Antioxidant compositions

Abstract

The present disclosure relates to an antioxidant composition, capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprising a hops extract and / or an extract comprising rosmarinic acid or combinations thereof. It further relates to a method of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff by using the antioxidant compositions and uses thereof.

Description

[0001] ANTIOXIDANT COMPOSITIONS

[0002] FIELD OF THE INVENTION

[0003] The present disclosure relates to antioxidant compositions capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprising a hops extract and / or an extract comprising rosmarinic acid. It further relates to a method of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, preferably a fermented beverage.

[0004] BACKGROUND

[0005] Fermented beverages and / or foodstuffs, especially beers, are one of the most consumed refreshments worldwide. Apart from the traditional beers, a lot of focus these days is also on the production and provision of non-alcoholic fermented beverages, especially non-alcoholic beers.

[0006] The typical beer brewing process comprises various steps. A non-exhaustive list of the individual steps includes: a. Malting: the process begins with malting, where grains (usually barley) are soaked in water and allowed to germinate, this activates enzymes that convert starches into fermentable sugars; b. Mashing: the malted grains are then crushed and mixed with hot water in a process called mashing. This allows the enzymes to break down the starches into sugars, creating a sweet liquid known as wort; c. Lautering: after mashing, the liquid wort is separated from the solid grain material in a process called lautering. This is typically done by passing the mixture through a filter bed, allowing the liquid to drain while retaining the grain solids; d. Boiling: the wort is then transferred to a large container and boiled. During this process, hops are added to the wort, contributing bitterness, flavor, and aroma to the beer. Boiling also helps sterilize the wort and coagulate proteins; e. Cooling: after boiling, the wort needs to be rapidly cooled to a temperature suitable for yeast fermentation. This can be achieved using a heat exchanger or by transferring the wort to a separate vessel and using a cooling system; f. Fermentation: once the wort is cooled, yeast is added to begin fermentation. The yeast consumes the sugars in the wort and produces alcohol and carbon dioxide as by products. Fermentation can take anywhere from a few days to several weeks, depending on the desired beer style; g. Conditioning: after fermentation, the beer is often conditioned to further develop its flavors and carbonation. This can involve transferring the beer to a secondary vessel and allowing it to age for a period of time: h. Filtration and carbonation: before packaging, the beer may undergo filtration to remove any remaining solids or yeast. Carbonation can be achieved naturally through the fermentation process or by adding carbon dioxide; i. Packaging: finally, the beer is packaged into bottles, cans, or kegs for distribution and consumption.

[0007] It is important to note that there are variations in the brewing process depending on the type of beer being produced and the preferences of the brewer. Especially during the mashing process, the presence of fatty acids leads to undesirable off-notes or off-flavors. Fatty Acids, a subcategory of lipids, comprises a range of related organic chemicals that includes fats, oils, and waxes. Their physical properties are largely determined by the length and degree of unsaturation of the hydrocarbon fraction of their molecular structures. The nonpolar hydrocarbon chain accounts for the poor solubility of fatty acids in water while the carboxylic acid group is polar and accounts for the higher solubilities of short-chain fatty acids in water. The simplest lipids constructed from fatty acids are triglycerides, better known as fats. Fatty acids are relevant in brewing because they negatively affect the organoleptic stability of beer. The breakdown of unsaturated fatty acids such as linoleic and linolenic acids or derivatives thereof or similar causative compounds into staling compounds such as (E)- 2-nonenal (i.e. trans-2-nonenal), hexanal, heptanal and 2,4-decadienal by oxidative mechanisms is well documented and gives rise to the typical off-note “cardboard flavor” in beers. This affects the taste perception of the beer and is highly undesirable.

[0008] Thus, slowing and / or preventing the oxidative degradation pathway of such fatty acids, especially linoleic and linolenic acids, provides a viable approach to prevent off-note generation in fermented beverages, especially beers.

[0009] Commonly used antioxidants like (bi)sulfites, ascorbic acid, commercial tannins may be used to prevent oxidative degradation of fatty acids. However, these components may need to be declared on the label or packaging or have other regulatory limitations, which creates another challenge for brewers looking for clean labelling of their fermented beverages. Another challenge for brewers is that the effectiveness of antioxidants may be greatly reduced when added to the fermentable medium before boiling. In other words, the antioxidant compositions need to maintain their effectiveness even after the brewing process, so that it can reduce and / or prevent off-note development, even after packaging.

[0010] Another option is to use naturally or synthetically occurring components having an antioxidative effect. Hop resins are divided between soft resins — those that are soluble in hexane — and hard resins — those that are soluble in ether. The soft resin fraction contains the hop’s alpha and beta acids, whereas the hard resin fraction contains the hop’s oxidized alpha and beta acids, as well as polyphenols. While hops are already used in beer for flavor and microbiological stabilization and is also known to be a source of polyphenols that could participate in reducing oxidation of the beer, these polyphenols are not desirable in high concentration because they can be responsible for unwanted haze in the finished product. This has limited the application of hops (extracts) to prevent oxidative processes leading to off notes in fermented beverages. Furthermore, the high concentrations of polyphenols, while being known in the art, may present solubility problems in aqueous systems like beers.

[0011] Other known alternative is the usage of components like rosemary extracts for oxidation prevention. However, this approach has not received much attention in the art, especially the compatibility and effectiveness of adding these extracts during beer brewing.

[0012] It is the aim of the present disclosure to overcome the limitations of the prior art. SUMMARY

[0013] In a first aspect the present disclosure relates to an antioxidant composition, capable of reducing and / or prevent off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprising a hops extract and / or an extract comprising rosmarinic acid or combinations thereof.

[0014] In a second aspect, the present disclosure relates to a method of reducing and / or preventing off- notes due to fatty-acid oxidation in a beverage and / or foodstuff, preferably a fermented beverage, comprising the steps of: a) providing an antioxidant composition as described herein; b) adding an effective amount of the antioxidant composition to the beverage and / or foodstuff.

[0015] In another aspect, the present disclosure relates to the use of the antioxidant composition described herein to reduce and / or prevent off-notes due to fatty-acid oxidation in a beverage and / or foodstuff.

[0016] It has been observed that the antioxidant compositions, as described herein, reduce and / or prevent the development of sensorially perceptible off-notes in a beverage and / or foodstuff that are derived from the oxidation pathways of fatty-acids, especially due to the breakdown of unsaturated fatty acids, such as linoleic acid, linolenic acid or derivatives thereof into staling compounds such as (E)-2- nonenal (i.e. trans-2-nonenal), hexanal, heptanal and 2,4-decadienal, especially trans-2-nonenal.

[0017] The reduced and / or prevented off-note or off-flavor maybe characterized as “papery”, “cardboardlike”, “oxidized” or in general “stale”.

[0018] DETAILED DESCRIPTION

[0019] The following text sets forth a broad description of numerous different embodiments of the present disclosure. The description is to be construed as exemplary only and does not describe every possible embodiment since describing every possible embodiment would be impractical, if not impossible. It will be understood that any feature, characteristic, component, composition, ingredient, product, step or methodology described herein can be deleted, combined with or substituted for, in whole or part, any other feature, characteristic, component, composition, ingredient, product, step or methodology described herein. Numerous alternative embodiments could be implemented, using either current technology or technology developed after the filing date of this patent, which would still fall within the scope of the claims.

[0020] The terms “comprises,” “comprising,” “has,” “having,” “includes,” “including,” “contains,” “containing,” or any other variation are open-ended and are intended to cover a non-exclusive inclusion of elements, such that an article, apparatus, compound, composition, combination, method, or process that “comprises,” “has,” or “includes,” or “contains” a recited list of elements does not include only those elements but may include other elements not expressly listed, recited or written in the specification or claims. An element or feature proceeded by the language “comprises . . .a,” “contains . . . a,” “has . . . a,” or “includes . . .a” does not, without more constraints, preclude the existence or inclusion of additional elements or features in the article, apparatus, compound, composition, combination, method, or process that comprises, contains, has, or includes the element or feature. The terms “a” and “an” are defined as one or more unless expressly stated otherwise or constrained by other language herein. An element or feature proceeded by “a” or “an” may be interpreted as one of the recited element or feature, or more than one of the element or feature.

[0021] The terms “about,” “approximately,” “essentially,” “substantially,” any other version thereof, or any other similar relative term, or similar term of approximation, are defined as being close to as understood by one having ordinary skill in the art. By way of non-limiting, illustrative embodiments, these terms are defined to be within 10% of recited value, or defined to be within 5% of a recited value, or defined to be within 4% of a recited value, or defined to be within 3% of a recited value, or defined to be within 2% of a recited value, or defined to be within 1 % of a recited value, or defined to be within 0.5% of a recited value, or defined to be within 0.25% of a recited value, or defined to be within 0.1% of a recited value.

[0022] It should be understood that when an amount in weight percent is described in the present disclosure, it is intended that any and every amount within the range, including the end points, is to be considered as having been expressly disclosed. For example, the disclosure of "a range of from about 1 to about 10" is to be read as indicating each and every possible number along the continuum between about 1 and about 10. It is to be understood that the inventors appreciate and understand that any and all data points within the range are to be considered to have been specified, and that the inventors have possession of the entire range and all points within the range.

[0023] When concentration is expressed as “ppm”, the concentration is parts per million by weight based on the total weight of the consumable or alternatively the “ppm” may also be construed as certain milligram per litre of a liquid consumable (mg / L). It should be understood that when a range of values is described in the present disclosure, it is intended that any and every value within the range, including the end points, is to be considered as having been disclosed. For example, “a range of from 1 ppm to 1000 ppm” of a component of the composition is to be read as indicating each and every possible number along the continuum between 1 and 1000. It is to be understood that the inventors appreciate and understand that any and all values within the range are to be considered to have been specified, and that the inventors have possession of the entire range and all the values within the range.

[0024] In a first embodiment, the antioxidant composition capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprises a hops extract and / or an extract comprising rosmarinic acid.

[0025] As used herein, the term “antioxidant composition” means a composition comprising an extract as described herein along with a suitable solvent, water and may further comprise other suitable components such as carriers, emulsifiers, stabilizers and the like. As used herein an “extract” means a composition comprising extractives (i.e. components extracted during the extraction process) and a solvent used during extraction and / or concentration of the extract. In a particular embodiment, the antioxidant composition and the extract may be used interchangeably.

[0026] As used herein, the term “capable of reducing and / or preventing off-notes due to fatty-acid oxidation” may be understood as the reduction and / or prevention of a sensorially perceptible off-note in a beverage and / or foodstuff that is derived from the oxidation of fatty-acids. The extracts, as described herein, impede the oxidation pathways and reduce and / or prevent the formation of off-note causing compounds, due to presence of anti-oxidative components in the extracts. The development of such compounds can be analytically determined e.g. using gas chromatography. The off-note can be identified either by trained panellists or a consumer of the beverage and / or foodstuff.

[0027] In a particular or preferred embodiment, the extracts as described herein reduce and / or prevent the breakdown of unsaturated fatty acids, such as linoleic acid, linolenic acid or derivatives thereof into staling compounds such as (E)-2-nonenal (i.e. trans-2-nonenal), hexanal, heptanal and 2,4-decadienal.

[0028] As used herein, the off-note or off-flavor maybe characterized as “papery”, “cardboard-like”, “oxidized” or in general “stale”. Such off-notes may develop in the fermented beverage, especially beer, and / or foodstuff shortly after packaging and keep on increasing thereafter. In a particular or preferred embodiment, the extracts, as described herein, reduce and / or prevent cardboard-like off-note developed in a beverage and / or foodstuff, especially fermented beverage. In a particular or preferred embodiment, the extracts, as described herein, reduce and / or prevent cardboard-like off-note developed due to the development of causative compounds like (E)-2-nonenal (i.e. trans-2-nonenal), hexanal, heptanal and 2,4- decadienal, especially in fermented beverage and more so in beer.

[0029] In a particular or preferred embodiment, the antioxidant composition may be in a liquid and / or solid form or even semi-solid form. The antioxidant composition may also be a suspension or in colloidal form.

[0030] It has been found that the antioxidant composition, comprising the extracts described herein, can be applied to reduce and / or prevent off-notes in a beverage and / or foodstuff, especially fermented beverage, more so beers, especially when added at the mashing stage of the brewing process. The antioxidant composition maintains its effectiveness in preventing or prolonging the development of off- notes, even after packaging.

[0031] Furthermore, said composition may be added without leading to any haze-development. As described herein, “haze” may be defined as the observance of turbidity resulting from undesired oxidative mechanisms, as described herein. Within the context of the present disclosure, haze was judged visually. It is surprisingly found that the antioxidant compositions, described herein, prevent the formation of detectable haze in the beverage and / or foodstuff. If needed, haze or turbidity in fermented beverages such as beers may also be determined according to the method of the American Society of Brewing Chemists (doi: 10.1094 / ASBCMOA-Beer-26). In a particular or preferred embodiment, the antioxidant composition as described herein does not lead to the formation of haze, when added during fermentation of a beverage, especially a beer and preferably when added during the mashing stage.

[0032] In a particular or preferred embodiment, the antioxidant composition described herein exhibits lower haze formation than a control, wherein the antioxidant composition is added to a beverage, preferably a fermented beverage, more preferably a beer. The haze formation or colloidal stability may be measured by any suitable method. On such method is EBC Analytica method 9.30.

[0033] In a particular or preferred embodiment, the antioxidant composition, as described herein, is present in the final product (such as the beverage and / or foodstuff) in at least about 100 ppm, preferably at least about 400 ppm, even more preferably at least about 500 ppm. In another particular or preferred embodiment, the antioxidant composition, as described herein, is present in the final product (such as the beverage and / or foodstuff) in at least about 100 ppm to about 1000 ppm, preferably from about 300 ppm to about 600 ppm, even more preferably from about 300 ppm to about 500 ppm. For clarification, the ppm amounts noted are the amount of the antioxidant composition in the final product. Thus, a skilled person such as a brewer or formulator can adjust the amount of the antioxidant composition in such a manner that eventually the noted ppm levels are obtained in the final product such as a beverage and / or foodstuff. As an example, the brewer may add more than 100 ppm at the mashing stage of the brewing process in order to obtain about 100 ppm in the final fermented beverage, especially beer. In a particular embodiment, the antioxidant composition, as described herein, is added in at least 500 ppm in case of hops extract and at least 300 ppm in case of an extract comprising rosmarinic acid, especially during the brewing of the fermented beverage and / or foodstuff, more especially during the mashing stage.

[0034] In a particular embodiment, the antioxidant composition may comprise or consist essentially of the extracts described herein, suitable solvent or carrier and water. The antioxidant composition may further comprise other suitable components such as carriers, emulsifiers, solvents, stabilizers and the like. In case of a solvent or carrier added to an antioxidant composition, this is over and above the solvent used in the extraction and / or concentration of an extract.

[0035] In particular embodiments, the carrier may be selected from the group consisting of, but not limited to, glycerin, ethanol, methanol propylene glycol, ethylene glycol, dipropylene glycol triacetin, maltodextrin, water and mixtures thereof. In another particular embodiment, the carrier may be selected from glycerin, water or combinations thereof. As used herein, the term “carrier” is the material that can be used to dilute or to dissolve the extract.

[0036] In another particular or preferred embodiment, the antioxidant composition, as described herein, capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprising a hops extract and / or an extract comprising rosmarinic acid provides a storage stable beverage and / or foodstuff.

[0037] By “storage stable” is meant that a beverage and / or foodstuff treated with the antioxidant composition, as described herein, prevents the development of undesirable off-notes due to fatty acid oxidation, especially “papery”, “cardboard-like”, “oxidized” or in general “stale”.

[0038] In a particular or preferred embodiment, the antioxidant composition reduces and / or prevents the perceptible undesirable off-note (i.e. is considered storage-stable) from developing at least for a period of 1 week, preferably 2 weeks, more preferably 3 weeks and even more preferably 4 weeks, from the time the beverage and / or foodstuff is packaged. In another particular or preferred embodiment, the antioxidant composition reduces and / or prevents the perceptible undesirable off-note from developing at least for a period of 1 month, preferably 3 months, more preferably 6 months and even more preferably 12 months, from the time the beverage and / or foodstuff is packaged. In a particular or preferred embodiment, the antioxidant composition reduces and / or prevents the perceptible undesirable off-note (i.e. is considered storage-stable) from developing or shows lower levels of detected trans-2-nonenal at least for a period of 1 week, preferably 2 weeks, more preferably 3 weeks and even more preferably 4 weeks, from the time the beverage and / or foodstuff is packaged when compared to known antioxidants like potassium metabisulfite or tannins. The storage stability may be determined as provided in the examples.

[0039] In a particular or preferred embodiment, said off-note is “cardboard-like”. The off-note can be sensorially detected by a trained flavorist or even a normal user consuming the beverage and / or foodstuff, especially a beer. The antioxidant composition capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprises a hops extract.

[0040] The hops used in the hops extract are derived from the hops plant, especially female flowers from the hops plant. It has been observed that hops extracts derived from the female flowers of the hops plants provide particularly effective extracts with antioxidative capabilities, containing xanthohumol and at least one other polyphenol.

[0041] In a particular or preferred embodiment, the hops extract is derived from spent hops. “Spent hops” are the remnants from supercritical CO2 extraction of hops, performed by hops processing companies to obtain liquid hop extracts. These spent hops are no longer useful for brewing and are typically discarded. However, it was discovered that these spent hops can be valorised for their antioxidative properties. In a preferred embodiment, the hops extract, as described herein, is derived from the spent hops, which are the remnants from supercritical CO2 extraction of hops, performed by hop processing companies to obtain liquid hop extracts. The spent hops may be in the form of pellets or another suitable form. The spent hops may have been already subjected to an extraction step, for e.g. using supercritical CO2 extraction.

[0042] Without being bound to theory, it is to be noted that the hops extract, as described herein, may be influenced by the extraction process of the starting (spent) hops material. As an example, the starting (spent) hops material, from which the hops extract is eventually obtained or obtainable, can be derived after extraction processes like supercritical CO2 extraction, hydrodistillation, alcoholic extraction etc. In a particular or preferred embodiment, the starting (spent) hops material is subjected to supercritical CO2 extraction.

[0043] As used in the present invention, the spent hops may be further extracted with a solvent. The material may be processed before extraction, for example it can be washed, dried, milled or grounded, etc. Particular solvents that may be used in the extraction process include water, alcohols (such as methanol, ethanol), acetone, ethyl acetate, hexane, dichloromethane, and any mixtures thereof, such as alcohol / water mixtures (such as mixtures of methanol and water). For example, the extraction solvents can be water, a water-alcohol mixture (from about 1 % to about 99% alcohol in water. For example, from about 30% to about 75% alcohol in water, or from about 30% to about 50% alcohol in water, such as from about 35% or from about 40% alcohol in water), or alcohol. Particular alcohols that may be mentioned include ethanol (EtOH) and methanol (MeOH).

[0044] In particular embodiments, the extraction solvent may be a methanol-water mix, such as from about 30% to about 90% methanol in water, or from about 30% to about 50% methanol in water. For example, from about 50% or from about 80% methanol in water. In a preferred embodiment, the extraction solvent is methanol-water mix with about 75% methanol and about 25% water. In another preferred embodiment, the extraction solvent may be an ethanol-water mix, preferably about 50% ethanol and about 50% water.

[0045] In a preferred embodiment of the composition of the invention the extraction of the hops material is carried out at a temperature of about 40°C to about 80°C, preferably about 50°C, for about 1 hour to about 3 hours, preferably about 2 hours.

[0046] In a preferred embodiment of the aerial parts, especially female flowers, obtained are further filtered and concentrated, preferably concentrated by membrane filtration. In a particular or preferred embodiment, the hops extract comprises at least xanthohumol and optionally at least one other polyphenol.

[0047] Xanthohumol, a prenylated flavonoid (a type of polyphenol) is found in the hard resin fraction of hops (humulus lupulus). It is somewhat unique, because nearly all other poly- and monophenols are found in non-lupulin plant material in the hop cone. Xanthohumol is the most abundant prenyl flavonoid in hops and may amount to nearly 1% of the hop-cone dry weight, depending on variety. Xanthohumol has no brewing value, because it does not contribute to or modify beer flavor. Despite the high levels of xanthohumol in some hops, its presence in beer is very low because it isomerizes rapidly during kettle boil. Traditionally hopped lager beer may have between 0 and 30 parts per billion (ppb) xanthohumol, whereas more heavily hopped ales and porters can have as much as 100 to 700 ppb. Hence, xanthohumol enriched hops extract have not been known to be used for its anti-oxidative effects to reduce and / or prevent off-notes in a beverage and / or foodstuff, especially fermented beverage, more so beers.

[0048] The hops extract may contain other components or extractives, such as but not limited to catechins, procyanidins, alpha acids, beta acids, polyphenols and mixtures thereof. A non-exhaustive list of different components in the hops extract, as described herein, may be found in the characterization data provided in the examples.

[0049] In a particular or preferred embodiment, the hops extract may contain polyphenols selected from the group consisting of procyanidins, quercetin, catechin, epicatechin and the like.

[0050] In a particular or preferred embodiment, the hops extract does not comprise any (added) tannins. For clarity, the hops extract, as described herein, may comprise unavoidable insignificant amounts of tannins, which may arise from the materials used to produce and / or process the antioxidant composition. The antioxidant composition not comprising tannins allows the use of a clean-label for beer manufacturers as tannins may need to be declared on the label as an added ingredient. Furthermore, it has been observed that the extracts, as described herein, effectively reduce and / or prevent off-notes even in the absence of tannins.

[0051] In a particular or preferred embodiment, the antioxidant composition may comprise at least about 30 wt.% of hops extractives, preferably at least 40 wt.%, even more preferably at least 50 wt.%. As used herein, “hops extractives” means a mixture of various compounds contained in the hops extract, as described herein, wherein the compounds comprise at least xanthohumol and optionally at least one another polyphenol. In a particular or preferred embodiment, the hops extract comprises at least about 1.5% w / w, at least about 1.7% w / w, at least about 1.8% w / w or even at least about 2% w / w of xanthohumol. In another embodiment, the hops extract comprises from about 0.5% w / w to about 5% w / w xanthohumol, preferably from about 1% w / w to about 3% w / w.

[0052] In another or alternative embodiment, the hops extract comprises from about 2 mg to about 5 mg, preferably from about 2.2 mg to about 5 mg, more preferably from about 2.5 mg to about 4.5 mg and even more preferably from about 2.5 mg to about 4 mg extracted xanthohumol per gram of spent hops material.

[0053] In another or alternative embodiment, the antioxidant composition comprises from about 0.5% to about 1 .5% w / w, preferably from 0.5% to about 1% w / w of xanthohumol.

[0054] In a particular or preferred embodiment, the antioxidant composition comprising the hops extract described herein exhibits lower haze formation than a control, wherein the antioxidant composition is added to a beverage, preferably a fermented beverage, more preferably a beer. The haze formation or colloidal stability may be measured by any suitable method. On such method is EBC Analytica method 9.30.

[0055] In another or alternative embodiment, the antioxidant composition capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprises an extract comprising rosmarinic acid.

[0056] The extract comprising rosmarinic acid may be obtained or derived from typical natural (botanical) sources of rosmarinic acid. These include, without limitations, selected from the group consisting of rosemary (rosmarinus officinalis), lemon balm (Melissa officinalis), sage (salvia officinalis), oregano (origanum vulgare), thyme (thymu vulgaris), basil (ocinum basilicum), perilla (perfilla frutescens) and mixtures thereof.

[0057] Rosmarinic acid is an ester formed via the esterification of caffeic acid with 3,4- dihydroxyphenyl lactic acid. Rosmarinic acid is defined in the entry of the database CAS Registry with the number 20283 92 5 as well as structurally as shown below:

[0058] The extract comprising rosmarinic acid used in the present invention may further comprise other components like one or more of Isomelitric Acid, Syringic Acid, Caffeoyl tartaric acid, Caffeic acid, yunnaneic acid derivative, Sagerinic acid / caffeic acid tetramer, Luteolin-7-glucuronide, Salvianolic acid B, pulmitric acid B / sagecoumarin, Trihydroxycinnamic acid derivative.

[0059] In certain embodiments, the antioxidant composition or extract comprising rosmarinic acid comprises rosemary extractives. “Rosemary extractives” as used herein means that the extract obtained from rosemary plant material comprises at least rosmarinic acid and at least one other component, as described herein. In certain embodiments, the extract comprising rosmarinic acid comprises rosemary extractives in about 20% w / w to about 40% w / w. In case of extracts in powder form, the rosemary extractives may range from about 20% w / w to about 80% w / w.

[0060] In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise at least 1% w / w rosmarinic acid, such as at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60% or at least 70% w / w of rosmarinic acid.

[0061] In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise at least 1% w / w Syringic Acid, such as at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60% or at least 70% w / w of Syringic Acid.

[0062] In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise at least 1% w / w Isomelitric Acid, such as at least 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60% or at least 70% w / w of Isomelitric Acid.

[0063] In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise between about 1% w / w, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70% w / w of rosmarinic acid to about 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1% w / w of rosmarinic acid In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise between about 1% w / w, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70% w / w of Syringic Acid to about 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1% w / w of Syringic Acid.

[0064] In certain embodiments, the extract comprising rosmarinic acid used in the present invention may comprise between about 1% w / w, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70% w / w of Isomelitric Acid to about 60%, 50%, 40%, 30%, 20%, 10%, 5%, 1% w / w of Isomelitric Acid.

[0065] In another embodiment, the effective amount of the extract comprising rosmarinic acid added to the beverage and / or foodstuff provides between about 1 ppm to about 1000 ppm of rosmarinic acid, such as from about 2ppm, 3ppm, 5ppm, 10 ppm, 20ppm, 30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm, 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, 600ppm, 700ppm, 800ppm, 900ppm to WOOppm, 900ppm, 800ppm, 700ppm, 600ppm, 500ppm, 400ppm, 300ppm, 200ppm, 100ppm, 90ppm, 80ppm, 70ppm, 60ppm, 50ppm, 40ppm, 30ppm, 20ppm, 10ppm, or to 5ppm. In certain embodiments, the beverage and / or foodstuff comprises from about 50 ppm of rosmarinic acid; preferably 1 ppm to about 50 ppm of rosmarinic acid, preferably about 1 ppm to about 40 ppm rosmarinic acid, more preferably about 1 ppm to about 30 ppm of rosmarinic acid, even more preferably about 10 ppm to about 15 ppm of rosmarinic acid in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0066] In another preferred embodiment, the beverage and / or foodstuff comprises between about 1 ppm to about 1000 ppm of rosmarinic acid, such as from about 2ppm, 3ppm, 5ppm, 10 ppm, 20ppm, 30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm, 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, 600ppm, 700ppm, 800ppm, 900ppm to WOOppm, 900ppm, 800ppm, 700ppm, 600ppm, 500ppm, 400ppm, 300ppm, 200ppm, 100ppm, 90ppm, 80ppm, 70ppm, 60ppm, 50ppm, 40ppm, 30ppm, 20ppm, 10ppm, or to 5ppm. In certain embodiments, the beverage and / or foodstuff comprises from about 50 ppm of rosmarinic acid; preferably 1 ppm to about 50 ppm of rosmarinic acid, preferably about 1 ppm to about 40 ppm rosmarinic acid, more preferably about 1 ppm to about 30 ppm of rosmarinic acid, even more preferably about 10 ppm to about 15 ppm of rosmarinic acid in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0067] In another embodiment, the beverage and / or foodstuff comprises between about 1 ppm to about 50 ppm rosmarinic acid, preferably about 1 ppm to about 40 ppm rosmarinic acid, more preferably about 1 ppm to about 30 ppm rosmarinic acid, even more preferably about 3 ppm to about 15 ppm in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0068] In another preferred embodiment, beverage and / or foodstuff comprises between about 1 ppm to about 1000 ppm of Syringic acid, such as from about 2ppm, 3ppm, 5ppm, 10 ppm, 20ppm, 30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm, 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, 600ppm, 700ppm, 800ppm, 900ppm to WOOppm, 900ppm, 800ppm, 700ppm, 600ppm, 500ppm, 400ppm, 300ppm, 200ppm, WOppm, 90ppm, 80ppm, 70ppm, 60ppm, 50ppm, 40ppm, 30ppm, 20ppm, Wppm, or to 5ppm. In certain embodiments, the beverage and / or foodstuff comprises from about 50 ppm of Syringic acid; preferably 1 ppm to about 50 ppm of Syringic acid, preferably about 1 ppm to about 40 ppm Syringic acid, more preferably about 1 ppm to about 30 ppm of Syringic acid, even more preferably about 10 ppm to about 15 ppm of Syringic acid in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0069] In another embodiment, the beverage and / or foodstuff comprises between about 1 ppm to about 50 ppm Syringic acid, preferably about 1 ppm to about 40 ppm Syringic acid, more preferably about 1 ppm to about 30 ppm Syringic acid, even more preferably about 3 ppm to about 15 ppm in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0070] In another preferred embodiment, the beverage and / or foodstuff comprises between about 1 ppm to about 1000 ppm of Isomelitric acid, such as from about 2ppm, 3ppm, 5ppm, 10 ppm, 20ppm, 30ppm, 40ppm, 50ppm, 60ppm, 70ppm, 80ppm, 90ppm, 100ppm, 200ppm, 300ppm, 400ppm, 500ppm, 600ppm, 700ppm, 800ppm, 900ppm to OOppm, 900ppm, 800ppm, 700ppm, 600ppm, 500ppm, 400ppm, 300ppm, 200ppm, 100ppm, 90ppm, 80ppm, 70ppm, 60ppm, 50ppm, 40ppm, 30ppm, 20ppm, 10ppm, or to 5ppm. In certain embodiments, the beverage and / or foodstuff comprises from about 50 ppm of Isomelitric Acid; preferably 1 ppm to about 50 ppm of Isomelitric acid, preferably about 1 ppm to about 40 ppm Isomelitric acid, more preferably about 1 ppm to about 30 ppm of Isomelitric acid, even more preferably about 10 ppm to about 15 ppm of Isomelitric acid in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0071] In another embodiment, the beverage and / or foodstuff comprises between about 1 ppm to about 50 ppm Isomelitric acid, preferably about 1 ppm to about 40 ppm Isomelitric acid, more preferably about 1 ppm to about 30 ppm Isomelitric acid, even more preferably about 3 ppm to about 15 ppm in respect to the final weight of the beverage and / or foodstuff or alternatively in respect to 1000 mL of the final beverage.

[0072] In one or preferred embodiment, the extract comprising rosmarinic acid is derived from lemon balm (Melissa officinalis) and / or rosemary (Rosmarinus officinalis). The common names and taxonomical names are interchangeable used in the present application.

[0073] In a preferred embodiment, the extract comprising rosmarinic acid is obtained from aerial parts. In certain embodiments, the extraction is performed using water and / or an alcoholic solvent. The term “alcoholic solvent” includes, without limitation, ethanol, methanol, heptane, and hexane. The term “aerial parts” as used herein refers to, but is not limited to, the shoots, leaves, stems, flowers, fruits and seeds, preferably the leaves or seeds of Melissa officinalis or Rosmarinus officinalis or any other botanical comprising rosmarinic acid.

[0074] In a preferred embodiment the aerial parts extraction is carried out with water and / or ethanol or mixtures of water and ethanol.

[0075] The material may be processed before extraction, for example it can be washed, dried, milled or grounded, etc. Particular solvents that may be used in the extraction process include water, alcohols (such as methanol, ethanol), acetone, ethyl acetate, hexane, dichloromethane, and any mixtures thereof, such as alcohol / water mixtures (such as mixtures of methanol and water). For example, the extraction solvents can be water, a water-alcohol mixture (from about 1 % to about 99% alcohol in water. For example, from about 30% to about 75% alcohol in water, or from about 30% to about 50% alcohol in water, such as from about 35% or from about 40% alcohol in water), or alcohol. Particular alcohols that may be mentioned include ethanol (EtOH) and methanol (MeOH). In particular embodiments, the extraction solvent may be a methanol-water mix, such as from about 30% to about 90% methanol in water, or from about 30% to about 50% methanol in water. For example, from about 50% or from about 80% ethanol in water. In a preferred embodiment, the extraction solvent is ethanol-water mix with about 75% methanol and about 25% water.

[0076] The term "acetone extract" as used herein, refers to the extract obtained from any member of the when the extraction from the plant (particularly, aerial parts) has been performed using acetone as the only solvent. The term "alcohol extract" as used herein, refers to the extract obtained when the extraction from the plant (particularly, leaves) has been performed using alcohol as the only solvent. For example, 100% methanol and / or 100% ethanol. The term "hydro-alcoholic extract" as used herein, refers to the extract obtained when the extraction from the plant has been performed using a mixture of water and alcohol. For example, from about 1 % to about 99% alcohol (e.g., ethanol, methanol) in water, such an extract would be termed a hydro-ethanolic extract.

[0077] A detailed procedure to prepare a Rosemary extract was described in the US Patent No. 5859293 (W096 / 34534), which is incorporated herein by reference in its entirety.

[0078] Water extractions are well known in the art and their application is part of the general knowledge of the expert. In a preferred embodiment of the composition of the invention the extraction of the aerial parts is carried out at a temperature of about 80°C to about 100°C, preferably 90°C, for about 1 hour to about 3 hours, preferably about 1 .5 hours, by percolation.

[0079] In a preferred embodiment of the aerial parts obtained are further filtered and concentrated, preferably concentrated by membrane filtration.

[0080] Once the concentration step has been carried out, the extract may be purified on polymeric adsorbent resin and the purified extract is spray dried on maltodextrin or formulated on ethanol or propylene glycol.

[0081] In another or preferred embodiment of the extract comprising rosmarinic acid from the aerial parts is obtained by an extraction with ethanol and activated carbon. In another preferred embodiment the of the composition of the invention the extract of said aerial parts is obtained with ethanol and activated carbon wherein the ethanol is used at a concentration of about 70% v / v solvent to water and the activated carbon is used at a concentration of about 1 % w / v to about 15% w / v, preferably about 3 % w / v, about 4 % w / v, about 5 % w / v, about 6 % w / v, about 7 % w / v, about 8 % w / v, about 10 % w / v.

[0082] In another embodiment, the material used for extracting the extract comprising rosmarinic acid can be any other part of the plant.

[0083] An exemplary process for extraction and isolation of extracts comprising rosmarinic acid of the invention may comprise the following steps:

[0084] (i) extraction of the plant parts, especially aerial parts, by a suitable solvent (such as acetone or ethanol);

[0085] (ii) evaporation of the solvent; and, if required

[0086] (iii) purification of the extract (e.g., by chromatography).

[0087] In certain embodiments, the temperature of extraction is in a range of from about 20 °C to about 100 °C. In a particular embodiment, the temperature for extraction is in a range of from about 50 °C to about 70 °C. Typically, the ratio of plant material to solvent mixture used in the extraction process varies from about 1 :1 to about 1 :10 on a gram to milliliter basis, such as from about 1 :3 to about 1 :8. The incubation period (i.e. the period during which the plant material is in contact with the solvent) is typically from about 2 hours to about 24 hours.

[0088] Mechanical energy can be applied during the extraction process. Applying mechanical energy helps to homogenize the mixture, changes the physical structure of the starting biological material and may increase the extraction yields. The amount of mechanical energy applied in the method depends on at which step it is applied, the type the material, the amount of the starting material used in the mixture, the pH of the mixture, and the temperature of the mixture. The amount of mechanical energy also can influence the amount of time needed to complete the extraction process.

[0089] For example, material (such as rosemary) and the extraction solution (such as acetone or ethanol) may be mixed using techniques known in the art, for example using stirring, maceration, percolation or infusion, such as magnetic or mechanical stirring.

[0090] Stirring may be conducted at any suitable revolution per minute (rpm), for example, the stirring may be done from about 1 rpm to about 10 rpm or about 50 rpm to about 500 rpm. For mechanical stirring, this may typically be done from about 1 rpm to 500 rpm, such as from about 10 rpm to about 200 rpm.

[0091] Devices for applying mechanical energy can be a pump, a refiner, a homogenizer, an extruder, a lobe pump, and / or a centrifugal pump. The mixture can be circulated in a closed-loop system that includes a pressure vessel (able to contain a heated solvent mixture), a reflux vessel, a heat exchanger, such as a shell and tube heat exchanger, and a pump for recirculating the heated mixture back to the vessel, allowing multiple passes through the pump in the system.

[0092] After the material (such as rosemary) and solvent have been incubated, the solvent is separated from residual material by any suitable separation technique known in the art (like for example filtration).

[0093] Further filtration steps can be used. The solvent may be partially or totally removed by any method known in the art such as centrifugation, Rota vap, and any device allowing solvent evaporation or a liquid-liquid way of replacing the solvent.

[0094] In certain embodiments, the Lamiaceae extract is enriched in phenolic diterpenes. As used herein, the term "Phenolic diterpenes" may refer to carnosic acid, carnosol, methylcar- nosate, and other phenolic diterpene derivatives (rosmanol, isorosmanol, 11 ,12-di-O-methyli- sorosmanol, 12-0- methylcarnosic acid, rosmanol-9-ethyl ether, circimaritin, Methylated monooxidized product of carnosic acid, genkwanin, epirosmanol, epiisorosmanol, carnosic acid derivative, epirosmanol ethyl ether, cryptotanshinone) and mixtures thereof.

[0095] In one embodiment, the extract obtained or obtainable from a plant of the Lamiaceae family, preferably rosemary or lemon balm, may be present in the antioxidant composition in an amount of from about 0.0001% to about 0.1%, such as from about 0.0005% to about 0.010% by weight of said composition. In another embodiment, the dry extract content (i.e. rosemary extractives) in the antioxidant composition may range from about 5 to about 40%.

[0096] In a particular or preferred embodiment, the antioxidant composition comprising the extract comprising rosmarinic acid, described herein, exhibits lower haze formation than a control, wherein the antioxidant composition is added to a beverage, preferably a fermented beverage, more preferably a beer. The haze formation or colloidal stability may be measured by any suitable method. On such method is EBC Analytica method 9.30. In a preferred embodiment, the present disclosure provides a method of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, preferably a fermented beverage, comprising the steps of: a) providing an antioxidant composition, as described herein; b) adding an effective amount of the antioxidant composition to the beverage and / or foodstuff.

[0097] As used herein, the term “effective amount” means the addition of the antioxidant composition in at least a minimum amount that is required to reduce and / or prevent off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, compared to said beverage and / or foodstuff not containing any antioxidant composition as described herein. The term may also mean that the addition of the antioxidant composition in at least a minimum amount to reduce and / or prevent sensorially perceptible off-note in a beverage and / or foodstuff that is derived from the oxidation of fatty-acids, compared to said beverage and / or foodstuff not containing any antioxidant composition as described herein. The off-notes may be characterized as “papery”, “cardboard-like”, “oxidized” or in general “stale”.

[0098] In a particular or preferred embodiment, the antioxidant composition added in an effective amount reduces and / or prevents the perceptible undesirable off-note (i.e. is considered storage-stable) from developing at least for a period of 1 week, preferably 2 weeks, more preferably 3 weeks and even more preferably 4 weeks, from the time the beverage and / or foodstuff is packaged. In another particular or preferred embodiment, the antioxidant composition reduces and / or prevents the perceptible undesirable off-note from developing at least for a period of 1 month, preferably 3 months, more preferably 6 months and even more preferably 12 months, from the time the beverage and / or foodstuff is packaged.

[0099] In a particular or preferred embodiment, the amount of the antioxidant composition added in an effective amount may mean that said composition is present in the final product (such as the beverage and / or foodstuff) in at least about 100 ppm, preferably at least about 400 ppm, even more preferably at least about 500 ppm. In another particular or preferred embodiment, the antioxidant composition, as described herein, is present in the final product (such as the beverage and / or foodstuff) in at least about 100 ppm to about 1000 ppm, preferably from about 300 ppm to about 600 ppm, even more preferably from about 300 ppm to about 500 ppm. For clarification, the ppm amounts noted are the amount of the antioxidant extract in the final product. Thus, a skilled person such as a brewer or formulator can adjust the amount of the antioxidant composition in such a manner that eventually the noted ppm levels are obtained in the final beverage and / or foodstuff. As an example, the brewer may add more than 100 ppm at the mashing stage of the brewing process in order to obtain about 100 ppm in the final fermented beverage, especially beer.

[0100] In another preferred embodiment, the present disclosure relates to a use of the antioxidant composition, as described herein, to reduce and / or prevent off-notes due to fatty-acid oxidation or to reduce haze formation in a beverage and / or foodstuff, especially a fermented beverage, more so a beer.

[0101] Broadly, foodstuffs, as used in the present disclosure, may be construed as of all kinds of confectionery products, baked products, sweet products, savoury products, fermented products, dairy products, beverages, oral care products, nutraceuticals and pharmaceuticals.

[0102] Exemplary foodstuffs include, but are not limited to, chilled snacks, sweet and savoury snacks, fruit snacks, chips / crisps, extruded snacks, tortilla / corn chips, popcorn, pretzels, nuts, other sweet and savoury snacks, snack bars, granola bars, breakfast bars, energy bars, fruit bars, other snack bars, meal replacement products, slimming products, convalescence drinks, ready meals, canned ready meals, frozen ready meals, dried ready meals, chilled ready meals, dinner mixes, meat analogs, frozen pizza, chilled pizza, soup, canned soup, dehydrated soup, instant soup, chilled soup, UHT soup, frozen soup, pasta, canned pasta, dried pasta, chilled / fresh pasta, noodles, plain noodles, instant noodles, cups / bowl instant noodles, pouch instant noodles, chilled noodles, snack noodles, dried food, dessert mixes, sauces, dressings and condiments, herbs and spices, spreads, jams and preserves, honey, chocolate spreads, nut-based spreads, and yeast-based spreads.

[0103] Exemplary confectionery products include, but are not limited to, chewing gum (which includes sugarized gum, sugar-free gum, functional gum and bubble gum), centerfill confections, chocolate and other chocolate confectionery, medicated confectionery, lozenges, tablets, pastilles, mints, standard mints, power mints, chewy candies, hard candies, boiled candies, breath and other oral care films or strips, candy canes, lollipops, gummies, jellies, fudge, caramel, hard and soft panned goods, toffee, taffy, liquorice, gelatin candies, gum drops, jelly beans, nougats, fondants, combinations of one or more of the above, and edible flavour compositions incorporating one or more of the above.

[0104] Exemplary baked products include, but are not limited to, alfajores, bread, packaged / industrial bread, unpackaged / artisanal bread, pastries, cakes, packaged / industrial cakes, unpackaged / artisanal cakes, cookies, chocolate coated biscuits, sandwich biscuits, filled biscuits, savoury biscuits and crackers, bread substitutes.

[0105] Exemplary sweet products include, but are not limited to, breakfast cereals, ready-to-eat (“rte”) cereals, family breakfast cereals, flakes, muesli, other ready to eat cereals, children's breakfast cereals, hot cereals.

[0106] Exemplary savoury products include, but are not limited to, salty snacks (potato chips, crisps, nuts, tortilla-tostada, pretzels, cheese snacks, corn snacks, potato-snacks, ready-to-eat popcorn, microwaveable popcorn, pork rinds, nuts, crackers, cracker snacks, breakfast cereals, meats, aspic, cured meats (ham, bacon), luncheon / breakfast meats (hotdogs, cold cuts, sausage), tomato products, margarine, peanut butter, soup (clear, canned, cream, instant, ultrahigh temperature “UHT”), canned vegetables, pasta sauces.

[0107] Exemplary dairy products include, but are not limited to, cheese, cheese sauces, cheese-based products, ice cream, impulse ice cream, single portion dairy ice cream, single portion water ice cream, multi-pack dairy ice cream, multi-pack water ice cream, take-home ice cream, take-home dairy ice cream, ice cream desserts, bulk ice cream, take-home water ice cream, frozen yoghurt, artisanal ice cream, dairy products, milk, fresh / pasteurized milk, full fat fresh / pasteurized milk, semi skimmed fresh / pasteurized milk, long-life / uht milk, full fat long life / uht milk, semi skimmed long life / uht milk, fat-free long life / uht milk, goat milk, condensed / evaporated milk, plain condensed / evaporated milk, flavoured, functional and other condensed milk, flavoured milk drinks, dairy only flavoured milk drinks, flavoured milk drinks with fruit juice, soy milk, sour milk drinks, fermented dairy drinks, coffee Whiteners, powder milk, flavoured powder milk drinks, cream, yoghurt, plain / natural yoghurt, flavoured yoghurt, fruited yoghurt, probiotic yoghurt, drinking yoghurt, regular drinking yoghurt, probiotic drinking yoghurt, chilled and shelf-stable desserts, dairy-based desserts, soy-based desserts. Exemplary beverages include, but are not limited to, flavoured water, soft drinks, fruit drinks, coffee-based drinks, tea-based drinks, juice-based drinks (includes fruit and vegetable), milk-based drinks, gel drinks, carbonated or non-carbonated drinks, powdered drinks, alcoholic or non-alcoholic drinks, and ready to drink liquid formulations of these beverages.

[0108] Exemplary fermented foods include, but are not limited to, cheese and cheese products, meat and meat products, soy and soy products, fish and fish products, grain and grain products, fruit and fruit products.

[0109] In a preferred embodiment, the beverage and / or foodstuff is a fermented beverage and / or foodstuff, especially a beer.

[0110] EXAMPLES

[0111] The following examples are provided solely for the purpose of illustration and are not to be construed as limitations of the present invention, as many variations of the invention are possible without departing from the spirit and scope of the present disclosure.

[0112] Materials

[0113] - Hops extract:

[0114] The raw material used is spent hop flower pellets (Hops supplier in Germany) from supercritical CO2 extraction. The material is ground and then extracted with ethanol 50% v / v for 2 hours at 50°C, with a solvent ratio where solvent volume is equal to 10 times the mass of raw material. Then, the native extract is filtered at a temperature above 40°C through an AF6 filter. The filtrate is concentrated under vacuum (50 mbar) until 10% of dry matter content is obtained. At that point, the corresponding amount of carrier is added to the extract in order to reach, after final vacuum concentration, the following composition: 30% dry hop extractives, 50% carrier (glycerine) and 20% water;

[0115] - Rosemary extract obtained from leaves;

[0116] - Beer type (Kolsch and Pils).

[0117] Analytical Equipment

[0118] Unless otherwise indicated, any chromatographic analyses were carried out using an Agilent 1290 series LC system coupled to an Agilent G6546 accurate mass QTOF with electrospray ionisation (ESI)-Jet Stream Technology. Separation was performed on a column Eclipse Plus C18 (250x 4.6mm; 5pm).

[0119] Unless otherwise indicated, any mass spectrometry (MS) data acquisition as well as qualitative analysis were performed by using Agilent Mass Hunter Workstation Software. The accurate mass data for the molecular ions were processed using the Agilent Mass Hunter Qualitative Analysis B.10.00 software (Agilent Technologies). This software provides a list of possible elemental formulae by using the Generate Molecular Formula form peak spectrum.

[0120] Unless otherwise indicated, GC-MS analysis was performed by Agilent 7890B GC & Agilent 5977 MSD (with the High Efficiency Source (HES). 1. Hops extract in beer

[0121] 1.1. Determining contents of hops extract

[0122] 100 mg (on dry basis) of sample was diluted in 10 mL of mixture (MeOH / FW; V / V = 9 / 1) and extracted on ultrasonic bath during 15 min and filtered on 0.2 pm regenerated cellulose. Then, the infusion was analysed by HPLC coupled to ESI-QTOF-MS in negative / positive ionisation mode as well as by UPLC-DAD in order to carry out a characterisation and quantification of the primary (i.e. not all) compounds. The primary identified components are listed below in table 01 : Table d :

[0123] A more detailed list of identified components is presented below in table 02: Table 02

[0124] 1.2. Antioxidant activity of hops extract

[0125] The antioxidative activity of the hops extract was determined by Oxygen Radical Absorbance Capacity (ORAC) analysis. A suitable method for the ORAC analysis is as below:

[0126] Determination of the antioxidant capability by ORAC was conducted on a fluorescence micro plate reader Tecan Infinite, following the procedure of Ou et al. 2013 (AOAC Official Method 2012.23 Total Antioxidant Activity). The method describes how to carry out the analysis for lipophilic and hydrophilic sample.

[0127] ORAC-Assay measures the capacity of a sample to bind oxygen radicals. Reactive oxygen species generators (AAPH) are added in parallel reactions that contain equal amounts of a fluorescent probe (Fluorescein sodium salt). Reactions contain either a buffer blank or antioxidant samples and standard (Trolox). The antioxidant capacity of a sample is the net difference between the area under the curve (AUC) of the sample and that of the blank. This net AUC of sample is compared to the net AUC of Trolox standard concentration in order to have the ORAC value of the sample expressed as pmol Trolox equivalent per gram (pmol TE / g).

[0128] The result obtained is proportional to the quantity of extract. A blend of products will give an ORAC value proportional on ORAC of each extract weighted by their proportion in the blend;

[0129] ORAC blend = (ORAC of extract alone * proportion of extract in the blend).

[0130] Example: For a blend of 30 % extract 1 at ORAC = 2000 pmol TE Zg and 70 % of extract 2 at ORAC = 700 pmol TE Zg, the expected ORAC value of the blend: (2000 x 0.3) + (700 x 0.7) = 1090 pmol TE / g

[0131] The results are indicated below in table 03:

[0132] Table 03:

[0133] 1.3. Antioxidant activity of hops extract in beers (Kolsch and Pits)

[0134] The antioxidant composition comprising the hops extract was used in beer during the brewing process. It is added in the mashing water, i.e. when the mash tun is being filled with hot water, before malt and other cereals are mixed in. This is to ensure optimum homogeneity and to slow down oxidation of fatty acids when malts are mixed with water. Said composition was added at a dosage of 0.05 % w / v of the final beer volume. The beer is then brewed according to standard recipes. The beer is packaged in cans.

[0135] The beers are aged in accelerated way: 1 month at 40°C in an incubator, in the dark.

[0136] The tested beers were analyzed by GC-MS for trans-2-nonenal content according to the following method. Prior to all analyses, the beer was degassed. This was done by putting a volume of beer (~300mL) in a 600 mL beaker, covering with a watch glass and placing in an ultrasonic bath. The bath was turned on and the beer was defoamed (1 to 3 minutes).

[0137] The headspace method for beer analysis is based on the previous Beer Core Headspace Analysis. This method used 4-fluorobenzaldehyde (CAS# 459-57-4) as an internal standard. The selected aldehydes were: hexanal (CAS# 66-25-1), heptanal (CAS# 111-71-7), trans-2-nonenal (CAS# 18829-56- 6), E2,Z4-decadienal (CAS# 25152-83-4), and E2,E4-decadienal (CAS# 25152-84-5). These aldehydes tend to be found in beer at a very low level (ppb).

[0138] A GC-MS Method with Selected Ion Monitoring (SIM) was used. A sample was prepared by pipetting 10mL of degassed beer into a headspace vial, adding 0.5g of sodium bicarbonate (CAS# 144- 55-8), add 50uL of the internal standard solution and then sealing the vial.

[0139] Internal Standard Solution: 4-fluorobenzadyde in ethanol (~10ppm).

[0140] GCMS: Agilent 7890B GC & Agilent 5977 MSD (with the High Efficiency Source (HES));

[0141] Autosampler: Gerstel MultiPurpose Sampler (MPS) with SPME Fiber Tool and Heated Agitator;

[0142] GC Column: DB-5, 30m x 0.25mm x 0.25um (Agilent # 122-5032);

[0143] SPME Fiber: Light Blue (CAR / PDMS) (Supleco # 57295-U);

[0144] GCMS Method: Beer SIM Method Blue Fiber.M

[0145] Inlet: Gerstel Cooled Injection System (CIS4);

[0146] Mode: Splitless (with SPME inlet liner and Septumless Sampling Head (SLH));

[0147] Temperature: 270°C;

[0148] Carrier Gas: Helium;

[0149] Pressure: 6.31 psi.

[0150] GC Parameters:

[0151] Initial Temperature: 40°C for 5 minutes

[0152] Rate: 4°C / min

[0153] Final Temperature: 280°C for 10 minutes

[0154] MSD Parameters :

[0155] Acquisition Mode: SIM / Scan

[0156] MSD Scan Range: 35 - 450

[0157] Threshold: 150

[0158] Number of SIM Groups: 5 (Group start and stop times determined by retention time of the aldehydes).

[0159] Group 1 : hexanal

[0160] Ions: 44, 56, 57, 72, & 82 (Dwell 200)

[0161] Group 2: heptanal

[0162] Ions: 44, 55, 57, 70, 86, & 96 (Dwell 200)

[0163] Group 3: 4-fluorobenzaldehyde Ions: 75, 95, 96, 123, 124, & 126 (Dwell 200)

[0164] Group 4: trans-2-nonenal

[0165] Ions: 57, 70, 83, 96, & 111 (Dwell 200)

[0166] Group 5: 2,4-decadienal

[0167] Ions: 81 , 95, & 152 (Dwell 400);

[0168] MS Source Temperature: 230°C;

[0169] MS Quad Temperature: 150°C.

[0170] Gerstel MPS Parameters:

[0171] Incubate: 15 minutes at 50°C with mixing (250 rpm);

[0172] Extract: 45 minutes at 50°C no mixing;

[0173] Desorb: 2 minutes in GC Inlet at 270°C.

[0174] Prior to use condition the SPME fiber at 300°C for 30 minutes.

[0175] The trans-2-nonenal content in the tested beer is shown in fig. 01 (Kolsch) and fig.02 (Pils). After accelerated aging (40°C / 1 month), the amount of trans-2-nonenal in the beer treated with hops extract in the mash is significantly lower than in the control beer that received no treatment. Without being bound to theory, the hops extract hinders the oxidation reactions, as described herein, at least during the mashing which resulted in less trans-2-nonenal precursors present in the wort after mashing, thereby leading to a decreased amount of trans-2-nonenal in the fresh beer.

[0176] To further validate the behaviour of trans-2-nonenal in a hops extract treated beer compared to a control beer, the trans-2-nonenal content was analyzed in ambient aged beer (Kolsch) for 11 months. The result is indicated in fig. 03. Trans-2-nonenal level in the beer at end of ambient shelf life was found to be significantly decreased compared to control.

[0177] 1.4. Sensory evaluation of hops extract in beer (Pils)

[0178] To validate the effectiveness of hop extract, sensory variations across the accelerated shelf life of the beer samples were evaluated by 8 trained panellists. They ranked differences between each aged beer and its cold stored counterpart, based on the following attributes over a period of 4 weeks: aroma, flavour, hoppy notes, malty notes, off-notes (cardboard, fatty, grassy), bitterness, acidity, mouthfeel and aftertaste.

[0179] The results are displayed in table 03. The study confirmed that observed analytical effect of hop extract can be translated into positive sensory impact on the aged beer.

[0180] Table 04:

[0181] ‘Significant difference between the sample and the reference; NSD: Non significant difference. 1 .5. Colloidal stability in beer (Pits)

[0182] Colloidal stability was measured according to the EBC-Analytica method 9.30 - Prediction of shelf-life of beer, which measures the haze formation after a forcing test. It was found that the beer containing the hops extract exhibited a higher colloidal stability compared to a control beer. This was shown by the fact that the haze is lower in the beer with the hops extract compared to the control (Fig. 05).

[0183] 2. Rosemary extract in beer

[0184] 2.1. Determining contents of the extract containing rosmarinic acid

[0185] An extract comprising rosmarinic acid was obtained from leaves of rosemary plant according to the procedure described in WO 96 / 35434.

[0186] The obtained extract was analysed for rosemary extractives and their amounts. 100 mg of extract in 20 mL volumetric flask, diluted with a mixture of watenmethanol, 1 :1 (v / v) then filtered through 0.2 pm RC filter for LC / MS or 0.45 pm for HPLC. The UV detector and the Q-TOF were used in series to analyze the sample. 2 mass lists with the main compounds described in literature were used to help for the identification. The instrumental parameters are as below: HPLC and HPLC-MS analytical conditions:

[0187] • Column: Zorbax Eclipse Plus C18 250x4.6mm 5pm - Agilent

[0188] • Column temperature: 30°C

[0189] • Flow rate: 1 mL / min (split after DAD for LC-MS)

[0190] • Detection at 280 nm, 350 nm and Q-TOF

[0191] • Injection volume: 5 pL

[0192] • Mobile phase: A: 0.1% formic acid in water, B: 0.1% formic acid in acetonitrile

[0193] Post run: 5 min

[0194] Q-TOF parameters:

[0195] • Each sample was analyzed in full scan positive and negative mode. Moreover, 200 pL of each sample were pooled in a vial to be analyzed by MS / MS in negative and positive mode with three collision energies (10, 30 60).

[0196] • m / z range: 80-1700

[0197] • Fragmentor = 175 V

[0198] The results are shown in table 05: Table 05:

[0199] 2.2. Antioxidant activity of the extract in beer (Pits)

[0200] The antioxidant activity and GC-MS determination of the aged beer (4 weeks, 40 °C) was performed in the same way as done for the hops extract (see before). The control beer was a Pils beer. The trans-2-nonenol content of the rosemary extract treated beer and control is shown in fig. 04. After accelerated aging, the amount of trans-2-nonenal in the beer treated with rosemary extract in the mash was observed to be lower than in the control beer that received no treatment. Without being bound to theory, it is supposed that the extract containing rosmarinic acid hinders the oxidation reactions, as described herein, during the mashing which resulted in less trans-2-nonenal precursors present in the wort after mashing, thus a decreased amount of trans-2-nonenal in the fresh beer.

[0201] 2.3. Sensory evaluation of rosemary extract in beer (Pils)

[0202] To validate the effectiveness of rosemary extract, sensory variations across the accelerated shelf life of the beer samples were evaluated by 8 trained panellists. They ranked differences between each aged beer and its cold stored counterpart, based on the following attributes over a period of 3 weeks: Aroma, Flavour, hoppy notes, malty notes, off-notes (cardboard, fatty, grassy), bitterness, acidity, mouthfeel and aftertaste.

[0203] The results are displayed in table 05. The study confirmed that observed analytical effect of rosemary extract can be translated into positive sensory impact on the aged beer.

[0204] Table 06:

[0205] ‘Significant difference between the sample and the reference; NSD: Non significant difference.

Claims

CLAIMS1. An antioxidant composition, capable of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, comprising a hops extract and / or an extract comprising rosmarinic acid or combinations thereof.

2. The antioxidant composition according any of the preceding claims, wherein said composition is in liquid and / or solid form.

3. The antioxidant composition according any of the preceding claims, wherein said hops extract does not comprise any tannin.

4. The antioxidant composition according to any of the preceding claims, wherein the hops is derived from spent hops.

5. The antioxidant composition according to any of the preceding claims, where the hops extract comprises xanthohumol and at least one another polyphenol.

6. The antioxidant composition according to any of the preceding claims, where the hops extract comprises: at least about 1 .5% w / w, at least about 1 .7% w / w, at least about 1 .8% w / w or even at least about 2% w / w of xanthohumol; and / or comprises from about 2 mg to about 5 mg, preferably from about 2.2 mg to about 5 mg, more preferably from about 2.5 mg to about 4.5 mg and even more preferably from about 2.5 mg to about 4 mg extracted xanthohumol per gram of spent hops material.

7. A beverage and / or foodstuff comprising the antioxidant composition according to any of claims 1 to 6.

8. The beverage according to claim 7, wherein the beverage is a fermented beverage, preferably a beer.

9. The beverage according to claim 7 comprising at least about 100 ppm, preferably at least about 400 ppm, even more preferably at least about 500 ppm of the antioxidant composition.

10. The beverage according to claim 7 comprising from about 100 ppm to about 1000 ppm, preferably from about 300 ppm to about 600 ppm, even more preferably from about 300 ppm to about 500 ppm of the antioxidant composition.

11. A beverage and / or foodstuff, preferably a fermented beverage, even more preferably a beer, according to claim 7, wherein the beverage exhibits lower haze formation than a control beverage not containing the antioxidant composition.2612. A method of reducing and / or preventing off-notes due to fatty-acid oxidation in a beverage and / or foodstuff, preferably a fermented beverage, comprising the steps of: a) providing an antioxidant composition according to claims 1 to 6; b) adding an effective amount of the antioxidant composition to the beverage and / or foodstuff.

13. The method according to claim 12, wherein the antioxidant composition is added in step b) in at least about 100 ppm, preferably at least about 400 ppm, even more preferably at least about 500 ppm.

14. The method according to claim 12, wherein the antioxidant composition is added in step b) from about 100 ppm to about 1000 ppm, preferably from about 300 ppm to about 600 ppm, even more preferably from about 300 ppm to about 500 ppm.

15. The method according to claim 12, wherein the antioxidant composition is added in step b) to fermented beverage, preferably a beer, and wherein said composition is added at the mashing stage.

16. Use of the antioxidant composition according to claims 1 to 6 to reduce and / or prevent off-notes due to fatty-acid oxidation or to reduce haze formation in a beverage and / or foodstuff.

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