Liquid colouring compositions

Abstract

The invention relates to a liquid colouring composition comprising carotenoids, a method of production for such compositions, food products and a manufacturing process to obtain the food products.

Description

[0001] LIQUID COLOURING COMPOSITIONS

[0002] TECHNICAL FIELD

[0003] The present invention relates to colouring compositions for the use in food and beverages, particularly for use in dairy and cheese products, a process for producing such colouring compositions, and it relates to food and beverage products comprising said compositions.

[0004] BACKGROUND

[0005] The cheese industry uses the natural food colouring called annatto, which is made from seeds of the Bixa orellana tree, to obtain certain colour hues in the curd characteristic for different types of cheese. However, annatto food colouring has a major disadvantage. When processing the cheese, the annatto colorant does not only affect the colour of the cheese curd but also the colour of the whey.

[0006] Several attempts have been made to replace annatto with other carotenoids, such as p- carotene.

[0007] GB2248170 (Hoffmann-La Roche) relates to p-carotene emulsified (and encapsulated) with caseinate thereby claiming that with the caseinate the p-carotene is better trapped in the cheese curd and not transferred to the whey fraction.

[0008] WO9304598 (Betatene) relates to carotenoid emulsions comprising a carotenoid in an oil solvent dispersed in an aqueous dispersion of a water-dispersible matrix with a stabilizer, wherein the stabilizer may be casein or caseinate.

[0009] WO9907233 (Rhodia) relates to producing a uniform coloration in cheese by associating a food grade colorant, such as annatto or p-carotene, with a carrier, such as caseinate, which is then dispersed within the milk source that is processed into a final cheese product.

[0010] EP1964479 (DSM IP Assets) relates to a powder containing carotenoids for the use in food and beverage compositions but not as a colouring composition but for health effects. Therefore, the aim was to provide a powder that does not have a noticeable colouring effect on the final water-based food or beverage product.

[0011] WO201 0069889 (Cybercolors) relates to a composition for cheese colouring comprising a composition of a paprika containing oil phase and a carotenoid and a fat in an aqueous phase comprising caseinate.

[0012] W02007003543 (BASF) relates to a process for manufacture of aqueous suspensions of carotenoids by grinding the carotenoid with modified starch and a sugar.

[0013] 34766- WO-PCT W02007009601 (DSM IP Assets) relates to solid or liquid water-dispersible colouring compositions containing carotenoids, preferably p-carotene, finely dispersed in a (modified) carbohydrate or (modified) protein carrier as colorants for food, beverages, or other applications.

[0014] WO201 0112406 (BASF) relates to ready-to-use stable suspensions of partially amorphous carotenoid particles used as a colouring composition for food and beverages with improved colour yields in the coloured products by providing finely dispersed particles of carotenoids in a protective colloid.

[0015] EP2717720 (Chr. Hansen) relates to colouring compositions for food, beverages and other applications comprising a carotenoid which is emulsified in an oil-in-water emulsion, and which further comprises solid carotenoid particles which are encapsulated in a hydrocolloid making it water-dispersible and thereby miscible with the oil-in-water emulsified carotenoid fraction.

[0016] EP4186370 (Chr. Hansen) relates to similar compositions as EP2717720 by further specifying the emulsifier of the oil-in-water phase and the encapsulating hydrocolloid used, whereas the hydrocolloid used for encapsulating the solid carotenoid particles is native casein.

[0017] In times of awareness of food waste, scarcity of resources, and whey having a lot of health benefits, it is not acceptable that the whey obtained from cheese making with annatto, - carotene, or other natural or nature-identical colorants, is discarded due to the colouring of the whey, or that further process steps of decolouration are required and thereby reduce sustainability. This is because coloured whey is not suitable for different food or health applications where the colour may be perceived as inapt. So, there is a need for colouring compositions that allow the curd to assume its colour whereas the whey remains uncoloured.

[0018] With EP2717720 and EP4186370, cheese colouring compositions were provided that generally achieve good colouring results of the curd and almost uncoloured whey. But the colouring compositions described therein have a major disadvantage over the coloration with annatto. The final colour of the curd only fully develops after several weeks of storage. With the fast-ripening techniques of the modern cheese maker this means that the cheese is ready for the market in terms of ripening, but the colour has not fully developed yet. A cheese product still going through colour changes might give rise to concern to consumers in view of potential food safety issues. Thus, a cheese that is ready for the market might not be sold due to such deferred colour development issue. There is no natural or nature-derived colorant available for cheese colouring, that does not leach into the whey and that does develop the final colour hue within the accelerated ripening times of 4 to 8 weeks.

[0019] 34766- WO-PCT But even annatto itself has a disadvantage as it often leads to some pinking of the curd after a certain time.

[0020] So, there is still a need for colouring compositions comprising carotenoids allowing the curd and / or the rind to assume its final colour hue at a reduced development time and whereas the whey obtained during cheese making with such colouring composition remains uncoloured. Especially, there is a need for colouring compositions whereas the colour of the curd and / or the rind is fully developed during ripening.

[0021] By combining different delivery concepts of the carotenoids, and p-carotene in particular, i.e., by combining liquid / liquid encapsulation with solid encapsulation and solubilizing both types of encapsulated particles in an isolated or purified milk protein or an isolated or purified milk protein extract, such as casein or caseinates, the inventors found a specific liquid colouring composition that overcomes the problems of the prior art and allows cheese making with an evenly coloured curd and an unchanged, thus colourless whey, and whereas the colour assumed by the curd is fully developed at the end of an even shortened ripening period of cheese of less than 12 weeks. A further advantage of combining the different concepts allows easy adjustment of the colour hue by varying the ratio of the different parts of the colouring composition.

[0022] SUMMARY

[0023] In a first aspect, a liquid colouring composition is provided for the colouring of a solid, a semisolid, or a liquid food, or a beverage comprising, i) a mixture of a) a liquid oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably said liquid oil-in-water emulsion does not contain a hydrocolloid, and b) a liquid aqueous dispersion of solid carotenoid particles, encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch, gelatine, gums, or pectins; and optionally a plasticizer and / or a filler, wherein the hydrocolloid is preferably a modified food starch, gelatine, gum acacia, or pectins, more preferably wherein the hydrocolloid is octenyl succinate anhydride starch (OSA-starch), and wherein preferably the carotenoid used to form said aqueous dispersion (part b) of the mixture) is crystalline, and

[0024] 34766- WO-PCT ii) a milk protein, or a milk protein extract, preferably an isolated milk protein or an isolated milk protein extract, more preferably casein or a caseinate, yet more preferably a sodium caseinate, a potassium caseinate, or a calcium caseinate, most preferably sodium caseinate or calcium caseinate.

[0025] In a second aspect of the present invention, a method of production of a liquid colouring composition for the colouring of a solid, a semi-solid, or a liquid food, or a beverage is provided comprising the steps of i) providing an oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably the emulsion is generated by dispersing the carotenoid in said oil, more preferably with the help of a solvent and / or a solubilizer, and emulsifying the carotenoid comprising said oil in an aqueous solution by homogenization and / or application of high-shear forces, ii) providing a liquid aqueous dispersion or suspension of solid carotenoid particles encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch and preferably a plasticizer, iii) blending, irrespective of the sequence of addition, said oil-in-water emulsion obtained in step i) with the liquid aqueous dispersion or suspension of solid carotenoid particles encapsulated in a hydrocolloid obtained in step ii), and with a milk protein, or a milk protein extract, preferably an isolated milk protein or an isolated milk protein extract, iv) optionally blending the composition from step i), ii), and / or iii) with additional ingredients, preferably a filler and / or antioxidants, more preferably said antioxidants being selected from the group consisting of ascorbic acid or salts thereof, tocopherol, citric acid, or mixtures thereof, and v) optionally homogenizing the so obtained liquid colouring composition, and wherein preferably said encapsulation of the carotenoid of step ii) is entailed by milling the liquid aqueous dispersion or suspension comprising the hydrocolloid with the solid crystalline carotenoid particles.

[0026] A third aspect of the invention is a method of making coloured cheese comprising the steps of i) adding a starter culture, rennet and the liquid colouring compositions as described hereinabove to a suitable milk base, ii) incubating the mixture under conditions favourable to the generation of the curd and whey fraction, iii) separating the curds from the whey.

[0027] 34766- WO-PCT A fourth aspect of the invention is a food product comprising said liquid colouring composition as described hereinabove.

[0028] A further aim of the present invention is to provide a use of said liquid colouring composition as described hereinabove for colouring cheese curd.

[0029] The details, examples and preferences provided in relation to any particular one or more of the stated aspects of the present invention will be further described herein and apply equally to all aspects of the present invention. Any combination of the embodiments, examples and preferences described herein in all possible variations thereof is encompassed by the present invention unless otherwise indicated herein, or otherwise clearly contradicted by context.

[0030] Brief Description of the Drawings

[0031] Figure 1 : CIE-L*a*b* values of different embodiments of the invention in comparison with DairyMax™ warm orange inside the cheese over 12 weeks showing similar L*a*b* values; a): L* inside, b): a* inside, c): b* inside

[0032] Figure 2: CIE-L*a*b values of different embodiments of the invention in comparison with DairyMax™ warm orange outside the cheese over 12 weeks showing different a* and b* values; a): L* outside, b): a* outside, c): b* outside

[0033] Figure 3: CIE-L*C*h° values of different embodiments of the invention in comparison with DairyMax™ warm orange inside the cheese over 12 weeks; showing similar C* and h° values for all formulations of Examples 5a-e; a): C* inside, b): h° inside

[0034] Figure 4: CIE-L*C*h° values of different embodiments of the invention in comparison with DairyMax™ warm orange in the cheese rind (outside) over 12 weeks; a): C* outside, b): h° outside; showing a significantly deteriorated colour in the cheese rind to a more greyish chroma in example 5e (DairyMax™)

[0035] Figure 5: A E values of different embodiments of the invention in comparison with DairyMax™ warm orange inside the cheese compared to within the cheese rind (outside) over 12 weeks; relating to a significantly deteriorated colour in the cheese rind to a more greyish chroma for example 5e (DairyMax™)

[0036] 34766- WO-PCT DETAILED DESCRIPTION

[0037] Throughout the present specification and the accompanying claims, the words “comprise, include” and “having” and variations such as “comprises , comprising, includes” and “including” are to be interpreted inclusively. That is, these words are intended to convey the possible inclusion of other elements or integers not specifically recited, where the context allows.

[0038] The articles “a” and “an” are used herein to refer to one or to more than one (i.e. to one or at least one) of the grammatical object of the article.

[0039] Carotenoids are a particularly useful source of colouring agents for a variety of foods and beverages. The carotenoids include natural or nature-identical carotenes (such as a-carotene, p-carotene, y-carotene, or mixtures thereof etc.), lycopene, bixin, zeaxanthin, cryptoxanthin, canthaxanthin, lutein, p-apo-8’-carotenal, p-apo-12’-carotenal and can provide colour pigments ranging from yellow to red. An especially important member of the carotenoid class is - carotene, because p-carotene is not only useful as a colorant (typically yellow, orange and especially red in colour) but also provides a valuable source of vitamin A having several health benefits. Most of the other carotenoids have also further beneficial properties in addition to their colouring capacity. For this reason, carotenoids are often included in foods and beverages as a colorant, especially where their further beneficial property, such as vitamin A fortification, is needed or desired.

[0040] It was therefore an object of the following invention to provide a liquid colouring composition comprising one or more carotenoids for cheese making, wherein the liquid colouring composition in a preferred case should colour the curd and rind but not have any noticeable colouring effect on the whey. Another objective of the invention was to provide a colouring composition for cheese that allows shortened development times of the coloration of the cheese curd and / or cheese rind to match shortened ripening times of modern cheese making.

[0041] Furthermore, the liquid colouring composition should satisfy the usual demands of a food composition, such as being stable against oxidation, being and staying evenly dispersed in the product overtime, and so on.

[0042] It has surprisingly been found that the object of the present invention is achieved by a liquid colouring composition for the colouring of a solid, a semi-solid, or a liquid food, or a beverage comprising i) a mixture of

[0043] 34766- WO-PCT a) a liquid oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably said liquid oil-in-water emulsion does not contain a hydrocolloid, and wherein the emulsion preferably comprises less than 5 wt-%, less than 4 wt-%, less than 3 wt-%, less than 2 wt-% of non-stabilized carotenoids (as crystalline filter residue), preferably less than 1 wt-%, more preferably less than about 0.9 wt-% of non-stabilized carotenoids (as crystalline filter residue), based on the total weight of the emulsion, and b) a liquid aqueous dispersion or suspension of solid carotenoid particles encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch, gelatine, gums, such as e.g. gum acacia, pectins, casein, or caseinates; and optionally a plasticizer and / or a filler, wherein the hydrocolloid is preferably a modified food starch, gelatine, gum acacia, or pectins, more preferably wherein the hydrocolloid is octenyl succinate anhydride starch (OSA-starch), and wherein preferably the carotenoid used to form said aqueous dispersion (part b) of the mixture) is crystalline, and ii) a milk protein, or a milk protein extract, preferably an isolated milk protein or an isolated milk protein extract, more preferably casein or a caseinate, yet more preferably a sodium caseinate, a potassium caseinate, or a calcium caseinate, most preferably sodium caseinate or calcium caseinate.

[0044] It was not to be foreseen by the person skilled in the art that the liquid colouring composition as described herein would solve the above-mentioned issues.

[0045] A preferred embodiment of the present invention is wherein said liquid oil-in water emulsion comprises a carotenoid, wherein the carotenoid is selected from the group consisting of natural or nature-identical carotenes, p-carotene, a-carotene, y-carotene, zeaxanthin, lutein, lycopene, or mixtures thereof. Any carotenoid may be present as different stereoisomers (cis / trans) and / or as mixtures thereof. Most preferred is a liquid oil-in-water emulsion wherein the carotenoid is a natural or a nature-identical carotene, preferably p-carotene.

[0046] A more preferred embodiment of the present invention is a liquid oil-in-water emulsion wherein the amount of the carotenoid ranges between about 1 wt-% and about 10 wt-%, more preferably the amount of the carotenoid ranges between about 1 wt-% and about 5 wt-%, most preferably

[0047] 34766- WO-PCT the amount of the carotenoid ranges between about 1 wt-% and about 3 wt-%-%, based on the total weight of the liquid oil-in-water emulsion, based on the total weight of the emulsion.

[0048] According to the present invention the “plant oil”, also called vegetable oil, that can be used but is not limited to is corn oil, sunflower oil, soybean oil, safflower oil, rape seed oil, arachis oil, palm oil, palm kernel oil, cotton seed oil, coconut oil, or MCT oil (medium-chain triglycerides). Preferred according to the present invention is corn oil, sunflower oil, coconut oil or MCT oil. More preferred to the present invention is corn oil.

[0049] When the one component (or mixture of components) is not (fully), i.e. partially, dissolved in the other component (or mixture of components), the composition is referred to as a “suspension” or “dispersion”.

[0050] Typically, dispersions are systems of particles dispersed in a continuous phase of a different material, wherein the particles and the continuous phase can be in the same or in a different state of matter.

[0051] A dispersion as referred to herein is to be understood as solid particles evenly dispersed in a liquid phase. The solid particles may be coated or encapsulated in a liquid or semi-liquid, such as a gel, and which is not or sparsely miscible with the continuous phase. In a preferred embodiment of the present invention, the dispersion is a liquid aqueous dispersion of particles relating to a system wherein the continuous phase is water.

[0052] In the present invention, a preferred dispersion is wherein the solid core is encapsulated in a liquid, semi-liquid, or gelled encapsulation agent, and more preferably wherein said encapsulation agent is a hydrocolloid as defined hereinbelow.

[0053] A suspension is a type of dispersion in which solid particles are dispersed throughout a liquid or gas but are large enough to eventually settle over time. Typical for a suspension is that particles are larger and less uniformly dispersed.

[0054] Suspensions and / or dispersions of the present invention are stabilized over a long period of time, preferably the suspensions or dispersions are stabilized for at least half a year, more preferably they are stabilized for between 6 months and 2 years, most preferably the suspensions or dispersions of the present invention are stabilized for between 10 and 16 months.

[0055] 34766- WO-PCT Emulsions are liquid particles dispersed in a non- or sparsely miscible liquid. Emulsions as referred to herein can also be particles having a solid core with a liquid surrounding them that is then dispersed in a non- or sparsely miscible liquid phase. Thus, the emulsion can be liquid particles in a non- or sparsely miscible liquid. A particular embodiment of the present invention are emulsions that contain liquid particles having a solid core dispersed in a non- or sparsely miscible liquid.

[0056] A preferred embodiment of the present invention is a liquid colouring composition wherein the amount of oil in the composition ranges between about 5 wt-% and about 40 wt-%, whereas the water content of the liquid colouring composition ranges between about 20 wt-% and about 45 wt-%, preferably the amount of oil in the liquid colouring composition ranges between about 15 wt-% and about 35 wt-%, based on the total weight of the liquid colouring composition.

[0057] Plasticizers as referred to herein are substances that increase plasticity and decrease viscosity of a material. Plasticizers that can be used in the present invention are selected from the group consisting of glycerol, sorbitol, citric acid, polyethylene glycol (PEG), or propylene glycol. Preferred plasticizers are glycerol and PEG, more preferred is glycerol. Plasticizers according to the present invention shall not include substances that entail a plasticizing effect only in a particular environment, such as sugars which exhibit a plasticizing effect in a biopolymer matrix such as starch or gelatin.

[0058] A preferred embodiment of the present invention relates to a liquid colouring composition wherein the plasticizer is glycerol or PEG. In a more preferred embodiment, the content of said plasticizer is at least 1 wt-% and ranges between about 1 wt-% and about 20 wt-%, preferably between about 1 wt-% and about 15 wt-%, more preferably between about 2 wt-% and about 10 wt-%, based on the total weight of the composition.

[0059] Another embodiment of the present invention relates to a liquid colouring composition wherein the plasticizer in part a) and part b) of the mixture may be identical or different. Preferably, the plasticizers in part a) and part b) of the mixture are identical.

[0060] A particularly preferred embodiment of the present invention relates to a liquid colouring composition wherein the liquid oil-in-water emulsion does not contain a hydrocolloid as defined hereinbelow.

[0061] Fillers as referred to hereinabove that may be added to the liquid aqueous dispersion or suspension may be maltodextrins, monosaccharides, such as fructose, glucose, maltose, disaccharides, such as lactose, maltose, sucrose, sugar alcohols, such as mannitol,

[0062] 34766- WO-PCT microcrystalline cellulose (MCC), cellulose, starches, calcium carbonate, dicalcium phosphate, or mixtures thereof, or preferably maltodextrins, fructose, glucose, or sucrose, or mixtures thereof.

[0063] The term “solid carotenoid particles” shall mean amorphous or crystalline particles of the respective carotenoid. In a preferred embodiment of the present invention said solid carotenoid particles are referred to as “solid core”. Embodiments comprising such solid carotenoid particles are explained hereinbelow.

[0064] A preferred embodiment of the present invention is wherein said liquid aqueous dispersion comprises a carotenoid, wherein the carotenoid is selected from the group consisting of natural or nature-identical carotenes, p-carotene, a-carotene, y-carotene, zeaxanthin, lutein, lycopene, or mixtures thereof. Any carotenoid may be present as different stereoisomers (cis / trans) and / or as mixtures thereof. Most preferred is a liquid aqueous dispersion wherein the carotenoid is p- carotene or natural carotenes comprising mixtures of p-carotene, a-carotene and other.

[0065] The carotenoid of the liquid aqueous dispersion (part b) of the mixture) may be the identical one or more carotenoid(s), or different one or more carotenoid(s) as the one or more carotenoid(s) of the liquid oil-in-water emulsion (part a) of the mixture). Preferred is an embodiment wherein the carotenoids in part a) and part b) of the mixture are identical, and wherein the one or more carotenoid(s) is / are carotene(s). More preferred is an embodiment, wherein said identical carotenoid is a natural or nature-identical carotene, most preferred wherein said identical carotene is p-carotene.

[0066] A more preferred embodiment of the present invention is a liquid aqueous dispersion (part b) of the mixture) wherein the amount of the carotenoid in said liquid aqueous dispersion ranges between about 1 wt-% and about 10 wt-%, more preferably the amount of the carotenoid ranges between about 1 wt-% and about 5 wt-%, most preferably the amount of the carotenoid ranges between about 1 wt-% and about 3 wt-%, based on the weight of the total weight of the liquid aqueous dispersion, based on the total weight of said liquid aqueous dispersion.

[0067] Preferably, the solid carotenoid particles used in the liquid aqueous dispersion (part b) of the mixture) of the present invention shall be generated from amorphous or crystalline particles, preferably they shall be generated from crystalline particles that have been milled before they are encapsulated.

[0068] 34766- WO-PCT The term “hydrocolloid” as used herein relates to polysaccharides, proteins or modified polysaccharides or proteins, or lignin sulfonates, or mixtures thereof.

[0069] The term "polysaccharide" as used herein includes acacia gum, pectins, celluloses, cellulose derivatives, and / or modified polysaccharides.

[0070] The term "modified polysaccharide" as used herein relates to polysaccharides which contain a lipophilic moiety, e. g. a hydrocarbon moiety having a chain length of preferably 5 to 18 carbon atoms in the straight chain. Preferably the modified polysaccharide should be acceptable for human consumption, i.e., preferred modified polysaccharides should be GRAS (generally recognized as safe) or approved for food consumption as determined by the various regulatory agencies worldwide. A preferred modified polysaccharide is modified food starch.

[0071] The term "modified food starch" as used herein relates to modified starches that are made from starches substituted by known chemical methods with hydrophobic moieties. For example, starch may be treated with cyclic dicarboxylic acid anhydrides such as succinic and / or glutaric anhydrides, substituted with an alkyl or alkenyl hydrocarbon group.

[0072] A particularly preferred modified starch of this invention has the following formula (I) wherein St is a starch, R is an alkylene radical and R’ is a hydrophobic group. Preferably R is a lower alkylene radical such as dimethylene or trimethylene. R’ may be an alkyl or alkenyl group, preferably having 5 to 18 carbon atoms. A preferred modified starch of formula (I) is starch sodium octenyl succinate ("OSA-starch").

[0073] The term "OSA-starch" as used herein denotes any starch (from any natural source such as corn, wheat, tapioca, potato or synthesized) that was treated with octenyl succinic anhydride (OSA). The degree of substitution, i.e. the number of esterified hydroxyl groups with regard to the total number of hydroxyl groups usually varies in a range of from 0.1 % to 10 %, preferably in a range of from 0.5 % to 5 %, more preferably in a range of from 2 % to 4 %.

[0074] 34766- WO-PCT OSA-starches are commercially available e.g. from National Starch under the trade names HiCap 100, Capsul, Capsul HS, Purity Gum 2000, UNI-PURE, HYLON VII; from Roquette Freres; from CereStar under the tradename C*EmCap or from Tate & Lyle.

[0075] The term “protein” as used herein relates to plant protein, such as protein from legumes such as pea, chickpea, beans, soybeans, or lentils; or proteins from grains such as wheat protein; or to animal protein, such as gelatine in hydrolysed or unhydrolyzed form, e.g., fish, swine, or bovine gelatine; or such as a milk protein or a milk protein, preferably an isolated milk protein or an isolated milk protein extract, more preferably extract such as whey protein, casein, or caseinates; egg protein; or a lignin sulfonate.

[0076] Encapsulated solid carotenoid particles shall mean that the carotenoid particles are fully engulfed by the encapsulating agent whereas no free-floating carotenoid particles can be observed.

[0077] Carotenoid particles encapsulated with a hydrocolloid comprised in the liquid aqueous dispersion according to the present invention as described herein are obtained by a process that requires that the solid, preferably crystalline carotenoid particles are dispersed in an aqueous medium and then milled together with the encapsulation agent, i.e., the hydrocolloid. In a preferred embodiment, the solid carotenoid particles used for the liquid aqueous dispersion are crystalline. In another preferred embodiment, the solid carotenoid particles used for the liquid aqueous dispersion comprise crystalline p-carotene or natural carotenes, comprising a mixture of p-carotene, a-carotene and other.

[0078] In a preferred embodiment said liquid colouring composition of the present invention comprises said oil-in-water emulsion comprising less than 5 wt-%, less than 4 wt-%, less than 3 wt-%, less than 2 wt-% of non-stabilized carotenoids (as crystalline filter residue) based on the total weight of the emulsion, preferably less than 1 wt-%, more preferably less than about 0.9 wt-% of non-stabilized carotenoids (as crystalline filter residue), based on the total weight of the emulsion. The content of said non-stabilized carotenoids can be determined as filtration residue e.g., by filtration through a suitable filter (such as paper, e.g., Whatman, pore size 4- 12pm with or without a filter aid and on a glass sinter) resulting as a fraction of the total weight of the emulsion.

[0079] The milk protein, or the milk protein extract, according to the present invention is a milk protein or a milk protein extract that has been isolated or purified, thus is an isolated or purified milk protein or an isolated or purified milk protein extract which can be selected from the group

[0080] 34766- WO-PCT consisting of native casein, or a casein salt, such as sodium, potassium or calcium caseinate.

[0081] Preferred are calcium or sodium caseinate. More preferred is sodium caseinate.

[0082] In a preferred embodiment said milk protein, or a milk protein extract, as defined above is contained in the liquid colouring composition in an amount ranging between about 0.01 wt-% and about 40 wt-%, more preferably in an amount ranging between about 1 wt-% and about 25 wt-%. Most preferably said milk protein, or a milk protein extract, as defined hereinabove is contained in an amount ranging between about 1 wt-% and about 15 wt-%, based on the total weight of the composition.

[0083] In an even more preferred embodiment, said milk protein, or said milk protein extract, as defined above, is attached to the carotenoid particles in the emulsion and the dispersion, but the particles of the emulsion and the dispersion are not encapsulated by said milk protein, or said milk protein extract. The inventors found that it is not necessary to encapsulate the carotenoid particles entirely with said milk protein, or said milk protein extract, adsorption on the surface is sufficient to make sure that the carotenoid particles stay in the cheese curd and do not leak colour into the whey. This effect may be even increased by reducing the size of the carotenoid particles in the liquid colouring composition as compared to carotenoid colouring compositions as described in the prior art.

[0084] In a preferred embodiment, the content of carotenoid in the liquid colouring composition based on the total weight of the composition ranges between about 0.1 wt-% to about 10 wt-%, more preferably between about 0.5 wt-% to about 5 wt-%, most preferably between about 1 wt-% to about 3 wt-%, based on the total weight of the composition.

[0085] In a preferred embodiment said optional plasticizer is contained in said liquid colouring composition in an amount ranging between about 1 wt-% to about 20 wt-%, more preferably in an amount ranging between about 1 wt-% to about 15 wt-%, most preferably the amount of the plasticizer in said liquid aqueous dispersion is between about 2 wt-% to about 10 wt-%, based on the total weight of the composition.

[0086] The liquid colouring composition according to the present invention entails a particle size distribution at d90 of below about 1200 nm, preferably below about 1000 nm, more preferably below about 850 nm and most preferably below 650 nm. Whereas the Sauter Mean Diameter d[3.2] is below about 0.3 pm, preferably between about 0.100 pm and about 0.300 pm, most preferably between about 0.100 and about 0.200 pm. The Sauter Mean Diameter (SMD) is a theoretical value that is best to describe particle sizes and particle size distributions of droplets

[0087] 34766- WO-PCT as it relates the volume to the surface area of particles. At the Sauter Mean Diameter the particle has the same ratio of volume to surface area as the entire ensemble of particles. A lower particle size is believed to ensure that said milk protein or said milk protein extract is adsorbed better on the surface of the encapsulated carotenoid particles leading to a better result in providing a coloured curd and clear whey.

[0088] The liquid colouring composition of the present invention as described hereinabove may further contain an emulsifier. Preferably the emulsifier is selected from the group consisting of non-ionic emulsifiers such as polysorbates, e.g., polyoxyethylene sorbitan, or such as citric acid esters, mono- / di-glycerides, or polyglycerol fatty acid esters. Preferred emulsifiers according to the present invention are polyoxyethylene sorbitan fatty acid esters or polyglycerol fatty acid esters, or mixtures thereof, and preferably in an amount of less than 5 wt-%, less than 4 wt-%, less than 3 wt-%, or less than 2 wt-%, more preferred in an amount of less than 1 wt-%, based on the total weight of the composition. Most preferred emulsifiers are polyglycerol fatty acid esters in an amount of less than 1 wt-%, based on the total weight of the composition.

[0089] A particularly preferred embodiment of the present invention is that said liquid colouring composition does not contain any emulsifier other than defined hereinabove, and more preferably the optional emulsifier added to the oil-in-water emulsion is not a hydrocolloid other than selected from the group as defined hereinabove, and most preferably the optional emulsifier added to the oil-in-water emulsion is not a hydrocolloid as defined hereinabove.

[0090] The antioxidant according to the present invention may be any water-soluble or fat-soluble antioxidant. Water-soluble antioxidants according to the present invention may be ascorbic acid and salts thereof, e.g., sodium ascorbate, and the like. The fat-soluble antioxidants may be a tocopherol, e.g., (all-rac)-tocopherol (i.e., synthetic tocopherol), (all-rac)-a-tocopherol (i.e., natural tocopherol), p- and y-tocopherol and mixtures thereof; butyl hydroxy toluene, butyl hydroxy anisol, propyl gallate, t-butyl hydroxy quinoline, or ascorbic acid esters of fatty acids such as ascorbyl palmitate or ascorbyl stearate. Depending on the pH of the aqueous matrix solution the latter two compounds may alternatively be added to the water phase. Preferred are combinations of a water-soluble and a fat-soluble antioxidant, more preferred is a combination of a tocopherol and ascorbic acid, or a salt or mixtures thereof.

[0091] In a preferred embodiment, the content of the emulsion of mixture a) and the content of the dispersion of mixture b) can vary to adjust and tailor the colour of the final composition which is required to adjust the colour to the respective cheese application. By adding more of the liquid aqueous dispersion of mixture b) the colour of the final composition turns more to an orange to

[0092] 34766- WO-PCT red hue. Preferably the ratio of the mixture of the emulsion a) to the dispersion of b) can vary in a range of between 99:1 and 1 :99, more preferably of between 10:1 and 1 :10, most preferably in a range of between 99:1 and 50:50.

[0093] The liquid colouring composition as described hereinabove can be manufactured according to the following method of production comprising the steps of i) providing an oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably the emulsion is generated by dispersing the carotenoid in said oil, more preferably with the help of a solvent and / or a solubilizer, and emulsifying the carotenoid comprising oil in an aqueous solution by homogenization and / or by application of high-shear forces, ii) providing a liquid aqueous dispersion or suspension of solid carotenoid particles encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch and preferably a plasticizer, iii) blending, irrespective of the sequence of addition, said oil-in-water emulsion obtained in step i) with the liquid aqueous dispersion or suspension of solid carotenoid particles encapsulated in a hydrocolloid obtained in step ii), and with a milk protein, or a milk protein extract, preferably an isolated milk protein, or an isolated milk protein extract, iv) optionally blending the composition from step i), ii) or iii) with additional ingredients, preferably a filler and / or antioxidants, more preferably said antioxidants being selected from the group consisting of ascorbic acid or salts thereof, such as preferably ascorbyl palmitate; tocopherols, such as preferably (all-rac)-a-tocopherol; citric acid, or mixtures thereof, and said fillers being selected from the group consisting of maltodextrins, monosaccharides, such as fructose, glucose, maltose, disaccharides, such as lactose, maltose, sucrose, sugar alcohols, such as mannitol, microcrystalline cellulose (MCC), cellulose, starches, calcium carbonate, dicalcium phosphate, or mixtures thereof, or more preferred are maltodextrins, fructose, glucose, or sucrose, or mixtures thereof; and v) optionally homogenizing the so obtained liquid colouring composition, and wherein preferably said encapsulation of the carotenoid of step ii) is done by milling the liquid aqueous dispersion or suspension comprising the hydrocolloid with the solid crystalline carotenoid particles.

[0094] In a preferred embodiment, the content of the emulsion of step i) and content of the dispersion of step ii) can be varied to adjust and tailor the colour of the final composition which is required to adjust the colour to the respective cheese application. Cheddar is more orange than continental cheeses like Gouda. By adding more of the dispersion of step ii) the colour of the

[0095] 34766- WO-PCT final composition turns more to an orange to red hue. Preferably the ratio of the mixture of the emulsion of step i) to the dispersion of step ii) can vary in a range of between 99:1 and 1 :99, more preferably of between 10:1 and 1 :10, most preferably in a range of between 99:1 and 50:50.

[0096] The liquid colouring composition of the present invention entails a higher colour intensity (E1 / 1) than what is available in the prior art. The E1 / 1 indicates the colour intensity of the colourant of a sample of 1 % solution and of 1 cm of thickness, thus corresponds to E (1 %, 1 cm). The higher the value the higher the colour intensity of the sample. The composition of the present invention achieves values of E1 / 1 of more than 1000. Preferably the composition of the present invention achieves values of E1 / 1 of more than 1030, more preferably of more than 1100, most preferably of more than 1150. The composition of the present invention can achieve values of E1 / 1 as high as about 2000.

[0097] The production of the liquid colouring composition as described hereinabove is provided in more detail.

[0098] Step i) of said method of production comprises manufacture of the oil-in-water emulsion as described hereinabove and may comprise the following steps: a) Dissolving the solid carotenoid particles and a plasticizer as defined hereinabove in a plant oil, and preferably a suitable solvent, and optionally adding a fat-soluble antioxidant, and optionally heating the solution. b) Homogenizing the obtained carotenoid solution in an aqueous solution of a plasticizer and optionally one or more additional ingredients such as an antioxidant, preferably a water- soluble antioxidant, c) After a homogeneous emulsion is obtained the solvent is removed by distillation, preferably under reduced pressure, preferably under a reduced pressure of below 400mbar.

[0099] Step ii) of said method of production comprises the manufacture of the liquid aqueous dispersion as described hereinabove and may comprise the following steps: a) Adding crystalline carotenoid particles to a solution of a plasticizer and a hydrocolloid in water, and optionally one or more additional ingredients such as a filler and / or one or more antioxidants, b) Homogenising the dispersion or suspension preferably by grinding / milling and / or with high- shear forces.

[0100] 34766- WO-PCT Step iii) of said method of production comprises the blending of the emulsion of step i) and the liquid aqueous dispersion of step ii) and a milk protein, or a milk protein extract both as defined herein above. The different liquids can be blended irrelevant of the sequence, i.e., it is rather irrelevant if both, the liquid oil-in-water emulsion and the liquid aqueous dispersion are blended with each other and then blended with said milk protein, or said milk protein extract, or if the emulsion and the dispersion are blended with said milk protein, or said milk protein extract, separately and thus said milk protein, or said milk protein extract containing emulsion and said milk protein containing dispersion, or said milk protein extract containing dispersion, are then blended. Preferably, when blending with said milk protein, or said milk protein extract, the blending should be without applying high-shear forces. For efficiency reasons, it is recommended and preferred blending the mixture of said emulsion of step i) and said dispersion of step ii) with the milk protein, or the milk protein extract, both as defined herein above.

[0101] It is not required that the carotenoid particles are encapsulated by the milk protein, or the milk protein extract, both as defined herein above. The inventors of the present invention have a theory as to why encapsulation by said milk protein, or said milk protein extract, is not required, but do not want to be bound by that theory: Said milk protein, or said milk protein extract, agglomerates at the outer layer of the carotenoid particles and helps to introduce the particles to the agglomerating forming cheese curd. It appears not to be required that the carotenoid particles are encapsulated by said milk protein, or said milk protein extract, adsorption on the surface appears to be sufficient.

[0102] According to optional step iv) the composition obtained in step iii) can be blended with one or more additional ingredients such as antioxidants, as defined hereinabove, stabilizers or fillers. Preferred are antioxidants comprising a mixture of a fat-soluble antioxidant and a water-soluble antioxidant, more preferred is a mixture of (all-rac)-a-tocopherol and ascorbic acid, salt thereof, or a mixture thereof. Preferred fillers may be maltodextrins, monosaccharides, such as fructose, glucose, maltose, disaccharides, such as lactose, maltose, sucrose, sugar alcohols, such as mannitol, microcrystalline cellulose (MCC), cellulose, starches, calcium carbonate, dicalcium phosphate, or mixtures thereof, or more preferred are maltodextrins, fructose, glucose, or sucrose, or mixtures thereof.

[0103] According to a preferred embodiment of the present invention, the additional ingredients, preferably the stabilizers or fillers, may be added to said milk protein, or said milk protein extract, before said milk protein, or said milk protein extract, is blended with the emulsion, the dispersion, or a blend of the emulsion and the dispersion as described hereinabove.

[0104] 34766- WO-PCT Preferred stabilizers or fillers are sucrose, salt, or mixtures thereof.

[0105] Homogenization as referred to herein is the process to break down and disperse particles in a liquid or semi-solid mixture in the emulsion, the dispersion or suspension to reduce the particle size and make a more uniform dispersion of the particles therein. Typical homogenization processes that can be used for the present invention are milling in a colloid mill (rotor-stator principle), or homogenizing in a high-pressure / high shear homogenizer, or milling in a wet grinding and dispersing equipment such as a bead mill or a ball mill. Preferably, a colloid mill (rotor-stator principle) is used for homogenization of the oil-in-water emulsion.

[0106] In another preferred embodiment, the liquid aqueous dispersion is milled in a wet milling process in a ball or bead mill which facilitates encapsulation and allows homogenization of the particles of the dispersion or the suspension, whereas the oil-in-water emulsion is preferably homogenized in a high-pressure / high-shear homogenizer or in a colloid mill (rotor-stator principle).

[0107] Suitable solvents for the use in step i) of the above method are organic solvents which preferably can be selected from the group consisting of halogenated solvents, such as chloroform, dichloromethane, water-miscible solvents, such as branched or unbranched alcohols, such as ethanol, isopropanol, isobutanol, water-immiscible solvents such as acetates, such as methyl acetate, ethyl acetate, n-butyl acetate, isopropyl acetate, isobutyl acetate, carbonates, such as dimethyl carbonate, diethyl carbonate, propylene carbonate, or ketones, such as acetone, methyl ethyl ketone, isobutyl methyl ketone, or mixtures thereof. More preferred solvents according to the method of the present invention are dichloromethane, ethanol, isopropanol, ethyl acetate, isopropyl acetate, isobutyl acetate, acetone or mixtures thereof.

[0108] Suitable solubilizers for the use in step i) of the above method can be lipids, non-ionic surfactants, organic liquids, such as beeswax or tocopherol, cyclodextrins or phospholipids or derivatives thereof, or mixtures thereof.

[0109] Cheeses which may be coloured according to this invention include, for example, Cheddar, Colby, Gouda, Monterey Jack and Leicester Cheese. It may be appreciated that any other natural cheese which is coloured for sale to consumers may be prepared with the novel carotenoid preparation.

[0110] 34766- WO-PCT Thus, the present invention is furthermore directed to a coloured cheese, particularly Cheddar, Gouda, Colby, Monterey Jack or Leicester Cheese, using and comprising the liquid colouring composition as defined hereinabove.

[0111] In cheese manufacturing, casein, fat, insoluble salts are reduced to a concentrated form by the action of rennet or acid or both. Pasteurized milk is inoculated with an appropriate starter culture to convert lactose to lactic acid, which lowers the pH of the milk. The novel carotenoid comprising liquid colouring composition, preferably a liquid colouring composition comprising natural or nature-identical p-carotene, is added and dispersed throughout the milk. At the starting pH of about 6.5 to about 6.8, the enzyme rennet is added to curdle the casein which precipitates and entraps as much as 90% of the milk fat. The amount of novel composition added to the milk depends on the colour of the cheese curd ultimately desired and is within the skill of the cheese producer. As a guidance, the range of novel composition added to milk is generally in the range from approximately 1 :5 000 to approximately 1 :40 000 (w / w).

[0112] Later, during the cheese manufacturing process, the curd is separated from the whey, suitably in conventional manner. The novel carotenoid comprising liquid colouring composition, preferably the novel liquid colouring composition comprising natural or nature-identical p- carotene, colours the cheese in the desired colour (i.e. yellow to orange colour for Cheddar Cheese) while the whey remains substantially uncoloured (white), or slightly greenish.

[0113] The following Examples illustrate the invention further without limiting it. All percentages and parts, which are given, are related to the weight and the temperatures are given in °C, and the pressures are absolute pressures when not stated otherwise.

[0114] EXAMPLES

[0115] Equipment used for characterization

[0116] Particle size distribution: p-carotene formulations were diluted in deionized water. Subsequently, particle size distributions were measured with a particle analyser Delsa™ Nano S (Beckmann Coulter, New Castle, DE, USA) using dynamic light scattering. Data was evaluated on the basis of intensity distributions.

[0117] Moreover, the laser diffraction instrument Mastersizer 3000 (Malvern Instruments, Malvern, UK) was used to assess volume-based particle size distributions. For the latter, particle and

[0118] 34766- WO-PCT dispersant refractive indices of 1 .596 and 1.330 were used, respectively. For emulsions the Mie scattering model for spherical particles was used.

[0119] Total carotenoid concentration

[0120] Total carotenoid concentrations of prototypes were determined by UV / Vis photometry (Lambda 365, PerkinElmer, Shelton, CT, USA).

[0121] In addition, the supernatant of the centrifuged stock solution was analyzed with HPLC-DAD analysis of total p-carotene. The Z-isomers of p-carotene were quantified on basis of the response of all-E p-carotene by correction with experimentally determined relative response factors.

[0122] Colour intensity (E1 / 1) and CIELAB( CIE-L*a*b*) colour values

[0123] The colour intensity E1 / 1 is defined as the difference in extinction at maximum wavelength and at 650 nm corrected for the concentration of carotenoid in the sample, amount weighted in and dilution. In detail, E1 / 1 is the absorbance of a sample of a 1% solution and of 1 cm of thickness and is calculated as follows: E1 / 1 (E (1 %, 1cm)) = (Amax-A650)*dilution factor I (weight of sample * content of product form in %).

[0124] Colour intensity (E1 / 1 - also called extinction coefficient) and CIE-L*a*b*, and CIE-L*C*h° respectively, colour values were measured using aqueous dispersions of p-carotene prototype formulations at total carotenoid concentrations of 5 ppm. An aliquot calculated based on the total carotenoid concentration (UVA / is photometry) was weighed into a 250 mL volumetric flask. The sample was further diluted to concentrations of 5 ppm. UVA / is spectra were recorded in the range of 300-700 nm. Absorbance was measured at the respective main absorption maximum (Amax) and at 650 nm (A650). Deionized water was used as reference. CIE-L*a*b colour values were recorded in total transmission mode (1cm, TTRANS; Colorimeter UltraScan Pro, HunterLab, Murnau, Germany).

[0125] Next to watery dispersions also dispersions in milk (Valflora UHT whole milk 3.5% fat) were made. A range of Oppm to 20ppm of total carotenoid concentrations were created. Dispersions were visually inspected and CIE-L*a*b* and CIE-L*C*h° colour values were measured. CIE- L*a*b and CIE-L*C*h° colour values were recorded in reflection mode (Colorimeter UltraScan Pro, HunterLab, Murnau, Germany).

[0126] CIE-L*a*b* colour values of the cheeses and the corresponding whey were measured in reflection mode. The values were converted into the CIE- L*C*h° by using following equations:

[0127] L* = L* h° = arctan ( — )

[0128] 34766- WO-PCT Filtration residue

[0129] The filtration residue represents an overall parameter to assess the quality of the formulated carotenoid, describing the carotenoid fraction that is not properly dispersed or comminuted and comprising an inner phase of bigger particle sizes, herein-called non-stabilized carotenoids. Aliquots of 600 mg - 1000 mg of formulation were re-dispersed in ca. 150mL deionized water and filtered through a filter paper (Whatman 589 / 2; 0 70mm, pore size 4-12 pm) and ~2.0 g filter aid (Hyflo) on a glass sinter. Subsequently, the filter was washed with deionized warm water until colourless water had passed through the filter. The carotenoid fraction remaining in the filter was recovered by stepwise washing it off the filter with dichloromethane, acetone as well as ethanol. The organic phase was collected and diluted with cyclohexane / ethanol 90:10 prior to recording UV / vis spectra in the range of 300-700nm. The absorbance was measured by UV / Vis photometry at the main absorption maximum using this same cyclohexane / ethanol 90:10 as blank. Filtration residues of non-stabilized carotenoids were calculated on the basis of the total carotenoid concentration (UV / Vis photometry) of the emulsion.

[0130] Example 1 : preparation of colouring composition

[0131] Step 1 a) p-carotene oil-in-water emulsion

[0132] A solution of 100g p-carotene, 220g corn oil, 15g (all-rac)-a-tocopherol and 50g of a polyglycerol ester of fatty acids (E475) in chloroform was homogenized into an aqueous solution of 400g glycerine and 20g ascorbyl palmitate (in 200g water) in a colloid mill (rotor-stator principle). The solvent was removed by distillation under reduced pressure (below 400mbar). The emulsion was filtered, and a filter residue of less than 1 wt-% of non-stabilized carotenoids was removed. The obtained emulsion was filled into suitable containers for storage.

[0133] Step 1 b) p-carotene dispersion

[0134] 100g crystalline p-carotene and 2g (all-rac)-a-tocopherol are added into a solution of 353g glycerol, 200g modified food starch (OSA-starch (E1450)), in 350g water. The suspension is homogenized and then ground in a bead mill. The obtained suspension was filled into suitable containers for storage.

[0135] 34766- WO-PCT Step 2 Blending p-carotene oil-in-water emulsion with p-carotene dispersion

[0136] The emulsion of step 1 a) (70 parts) and the dispersion of step 1 b) (30 parts) were blended and stirred for 30 minutes.

[0137] Step 3 Blending the mixture of step 2 with a caseinate solution

[0138] The mixture obtained in step 2 was then blended with a caseinate solution (sodium caseinate (Sodium Caseinate, Emulac Na, Meggle GmbH & Co. KG, Wasserburg, Germany 8.1 wt-%) and stirred for 30 minutes.

[0139] Example 2: preparation of colouring composition

[0140] Step 1 a) p-carotene oil-in-water emulsion

[0141] A solution of 100g p-carotene, 220g corn oil, 15g (all-rac)-a-tocopherol and 50g of a polyglycerol ester of fatty acids (E475) in chloroform was homogenized into an aqueous solution of 400g glycerine and 20g ascorbyl palmitate (in 200g water) in a colloid mill (rotor-stator principle). The solvent was removed by distillation under reduced pressure (below 400mbar). The emulsion was filtered, and a filter residue of less than about 0.8 wt-% of non-stabilized carotenoids was removed. The obtained emulsion was filled into suitable containers for storage.

[0142] Step 1 b) p-carotene dispersion

[0143] 100g crystalline p-carotene and 2g (all-rac)-a-tocopherol are added into a solution of 353g glycerol, 200g modified food starch (OSA-starch, (E1450)), and 350g water. The suspension was homogenized and then ground in a bead mill. The obtained suspension is filled into suitable containers for storage.

[0144] Step 2 Blending each of the mixtures of step 1 with a caseinate solution a) The emulsion of step 1 a) was blended with a caseinate solution ((4.05 wt-% Calcium Caseinate, Emulac Ca, Meggle GmbH & Co. KG, Wasserburg, Germany) and stirred for 30 minutes. b) The dispersion of step 1 b) was blended with a caseinate solution ((4.05 wt-% Calcium Caseinate, Emulac Ca, Meggle GmbH & Co. KG, Wasserburg, Germany) and stirred for 30 minutes.

[0145] 34766- WO-PCT Step 3 Blending the p-carotene oil-in-water-caseinate emulsion with the p-carotene-caseinate dispersion

[0146] The emulsion of step 2 a) (70 parts) and the dispersion of step 2 b) (30 parts) were blended and stirred for 30 minutes.

[0147] Example 3: preparation of colouring composition

[0148] Step 1 a) p-carotene oil-in-water emulsion

[0149] A solution of 100g p-carotene, 220g corn oil, 15g (all-rac)-a-tocopherol and 50g of a polyglycerol ester of fatty acids (E475) in chloroform was homogenized into an aqueous solution of 400g glycerine and 20g ascorbyl palmitate (in 195g water) in a colloid mill (rotor-stator principle). The solvent is removed by distillation under reduced pressure (below 400mbar). The emulsion was filtered, and a filter residue of less than about 0.9 wt-% of non-stabilized carotenoids was removed. The obtained emulsion is filled into suitable containers for storage.

[0150] Step 1 b) p-carotene dispersion

[0151] 100g crystalline p-carotene and 2g (all-rac)-a-tocopherol are added into a solution of 353g glycerol, 200g modified food starch (OSA-starch, (E1450)), in 345g water. The suspension was homogenized and then ground in a bead mill. The obtained suspension is filled into suitable containers for storage.

[0152] Step 2 Blending p-carotene oil-in-water emulsion with p-carotene dispersion

[0153] The emulsion of step 1 a) and the dispersion of step 1 b) were blended and stirred for 30 minutes.

[0154] Step 3 Blending the mixture of step 2 with a caseinate solution

[0155] The mixture obtained in step 2 was then blended with a caseinate solution (calcium caseinate (Calcium Caseinate, Emulac Ca, Meggle GmbH & Co. KG, Wasserburg, Germany 8.1 wt-%) and stirred for 30 minutes.

[0156] 34766- WO-PCT Example 4: preparation of colouring composition

[0157] Step 1 a) p-carotene oil-in-water emulsion

[0158] A solution of 100g p-carotene, 220g corn oil, 15g (all-rac)-a-tocopherol and 50g of a polyglycerol ester of fatty acids (E475) in chloroform was homogenized into an aqueous solution of 400g glycerine and 20g ascorbyl palmitate (in 195g water) in a colloid mill (rotor-stator principle). The solvent is removed by distillation under reduced pressure (below 400mbar). The emulsion was filtered, and a filter residue of less than about 0.8 wt-% of non-stabilized carotenoids was removed. The obtained emulsion is filled into suitable containers for storage.

[0159] Step 1 b) p-carotene dispersion

[0160] 100g crystalline p-carotene and 2g (all-rac)-a-tocopherol are added into a solution of 353g glycerol, 200g modified food starch (OSA-starch, (E1450)), in 345g water. The suspension was homogenized and then ground in a bead mill. The obtained suspension is filled into suitable containers for storage.

[0161] Step 2 Blending p-carotene oil-in-water emulsion with p-carotene dispersion

[0162] The emulsion of step 1 a) (70 parts) and the dispersion of step 1 b) (30 parts) were blended and stirred for 30 minutes.

[0163] Step 3 Blending the mixture of step 2 with a caseinate solution

[0164] The mixture obtained in step 2 was then blended with a caseinate solution ((Sodium Caseinate, Lactonat RN-5, Lactoprot Deutschland GmbH, Kaltenkirchen, Germany) and stirred for 30 minutes.

[0165] Example 5: Cheese making using the colouring composition of example 1-4 and the colorant DairyMax™ Warm Orange from Oterra

[0166] Cheddar type cheese:

[0167] - Jacket of the cheese vat has been filled with water and set to a temperature of 30.5°C

[0168] - Cheese vat is filled with pasteurized milk

[0169] - Starter cultures and colouring composition have been added and stirred.

[0170] - Rennet and CaCI2have been added and stirred

[0171] 34766- WO-PCT - Coagulum has been cut and under continuous stirring temperature was increased to 37.5°C.

[0172] - Stirring was continued until the pH gets to 6.2. Then stirring was stopped and the whey was drained. - When the pH is 5.3-5.4, the curd was milled.

[0173] - Salt was added (645g / 175L) and the milled curd was milled thoroughly

[0174] - The curd was placed into moulds and pressed. (Cheddar moulds, 3 bar), and then overnight at 4 bars.

[0175] - The pressed cheese was vacuum packed and ripened at 7°C. Table 1 : Overview of cheeses made in example 5 listing the different colorants / examples used

[0176] Example 6: Coloration data for different prototypes according to the present invention in examples 5a, c, and d compared to DairyMax™ (example 5e).

[0177] Table 2: CIE-L*a*b, CIE-L*C*h° and AE values of cheeses prepared according to examples

[0178] 5 a - d, and of the cheese preparation according to example 5 e: compared are the values for the cheese inside and the cheese rind (outside) during ripening.

[0179] 34766- WO-PCT

[0180] 34766- WO-PCT

[0181] The cheeses made using the colorants according to Examples 5 a-d show a significant advantage over the prior art in the Chroma (CIE-L*C*h°) and in the a* and b* values of the CIE- L*a*b* system, which translates to a brighter and more intense colour. Whereas the cheese using the DairyMax™ (example 5e) obtains a more greyish and duller colour after 12 weeks of ripening, which would not be appealing to a customer.

[0182] 34766- WO-PCT

Claims

Claims1 . A liquid colouring composition for the colouring of a food or a beverage comprising, i) a mixture of a) a liquid oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably said liquid oil-in-water emulsion does not contain a hydrocolloid, and b) a liquid aqueous dispersion of solid carotenoid particles, encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch, gelatine, gums, or pectins; and optionally a plasticizer and / or a filler, wherein the hydrocolloid is preferably a modified food starch, gelatine, gum acacia, or pectins, more preferably wherein the hydrocolloid is octenyl succinate anhydride starch (OSA-starch), and wherein preferably the carotenoid used to form said aqueous dispersion (part b) of the mixture) is crystalline, and ii) a milk protein, or a milk protein extract, preferably an isolated milk protein, or an isolated milk protein extract, more preferably casein or a caseinate, yet more preferably a sodium caseinate, a potassium caseinate, or a calcium caseinate, most preferably sodium caseinate or calcium caseinate.

2. The liquid colouring composition according to claim 1 , wherein the carotenoid in part a) and part b) may be identical or different, preferably the carotenoid is a carotene, more preferably the carotenoid is identical in part a) and b) and the carotenoid is natural carotene or nature-identical p-carotene.

3. The liquid colouring composition according to any one of the previous claims, wherein the oil-in-water emulsion comprises less than 5 wt-%, preferably less than 4 wt-%, less than 3 wt-%, less than 2 wt-% of non-stabilized carotenoids (as crystalline filter residue), more preferably less than 1 wt-%, most preferably less than about 0.9 wt-% of non-stabilized carotenoids (as crystalline filter residue), based on the total weight of the emulsion.

4. The liquid colouring composition according to any one of the previous claims, wherein the plasticizer is glycerol or PEG (polyethylene glycol), and wherein preferably the glycerol is comprised in an amount of at least 1 wt-%, preferably it ranges between about 1 and about34766- WO-PCT20 wt-%, more preferably between about 1 wt-% and about 15 wt-%, most preferably between about 2 wt-% and about 10 wt-%, based on the total weight of the composition.

5. The liquid colouring composition according to any one of the previous claims, wherein the particles of the liquid colouring composition have a particle size distribution d90 of less than about 1200 nm, preferably of less than about 1000 nm, more preferably of less than about 800 nm, even more preferably below 650 nm, and most preferably wherein the particles of the liquid colouring composition have a Sauter Mean Diameter (SMD; d[3.2]) of below about 0.3 pm.

6. The liquid colouring composition according to any of the previous claims, wherein the liquid oil-in-water emulsion (part a) of the mixture) further comprises an emulsifier in an amount of less than 1 wt-%, and wherein preferably the emulsifier is not a hydrocolloid.

7. The liquid colouring composition according to any of the previous claims, wherein the composition further comprises an antioxidant selected from the group consisting of tocopherol or ascorbic acid, or a salt thereof, or a mixture thereof.

8. A method of production of a liquid colouring composition comprising the steps of i) providing an oil-in-water emulsion comprising a carotenoid, a plant oil, a plasticizer, and water, wherein preferably the emulsion is generated by dispersing the carotenoid in said oil, more preferably with the help of a solvent and / or a solubilizer, and emulsifying the carotenoid comprising said oil in an aqueous solution by homogenization and / or by application of high-shear forces, ii) providing a liquid aqueous dispersion of solid carotenoid particles encapsulated in a hydrocolloid selected from the group consisting of modified or unmodified food starch, and preferably a plasticizer, iii) blending, irrespective of the sequence of addition, said oil-in-water emulsion obtained in step i) with the liquid aqueous dispersion of solid carotenoid particles encapsulated in a hydrocolloid obtained in step ii), and with a milk protein, or a milk protein extract, preferably an isolated milk protein, or an isolated milk protein extract, iv) optionally blending the composition from step i), ii), and / or iii) with additional ingredients, preferably a filler and / or antioxidants, more preferably said antioxidants being selected from the group consisting of ascorbic acid or salts thereof, tocopherol, citric acid, or mixtures thereof, and v) optionally homogenizing the so obtained liquid colouring composition, and34766- WO-PCTwherein preferably said encapsulation of the carotenoid of step ii) is entailed by milling the liquid aqueous dispersion of step ii) comprising the hydrocolloid with the solid crystalline carotenoid particles.

9. The method according to claim 8, wherein the carotenoid in step i) and ii) is a carotene, preferably p-carotene.

10. The method according to any one of the claims 8 or 9, wherein the antioxidant is selected from the group consisting of ascorbyl palmitate and tocopherol, or mixtures thereof.

11. Use of the liquid colouring composition according to any one of the claims 1 to 7 or obtained by the method according to any one of the claims 8 to 10 for colouring cheese curd.

12. A method of making coloured cheese comprising the steps of: i) adding a starter culture, rennet, and the liquid colouring composition according to any one of the claims 1 to 7 or obtained by the method according to any one of the claims 8 to 10 to a suitable milk base, ii) incubating the mixture under conditions favourable to the generation of the curd and whey fraction, iii) separating the curds from the whey.

13. A food product comprising the liquid colouring composition according to any one of the claims 1 to 7 or obtained by the method according to any one of the claims 8 to 10.

14. A food product according to claim 13 which is a cheese, preferably a Cheddar Cheese, a Gouda Cheese, a Monterey Jack, or a Leicester Cheese.34766- WO-PCT

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