Thermally stable oil in water emulsion

Abstract

Thermally stable oil-in-water emulsions including an oil containing polyunsaturated fatty acids. The present invention relates to thermally stable oil-in-water emulsions comprising an oil consisting of a polyunsaturated fatty acid, water, an emulsifier and a water-soluble stabilizer, and to processes for preparing thermally stable oil-in-water emulsions. The thermally stable oil-in-water emulsions remain flowable at a temperature of -40°C. <198> The particles do not change size after 9 months of storage at a temperature of -40°C. <198> ca -15 <198> w.

Description

1 / 80 THERMALLY STABLE OIL-IN-WATER EMULSION BACKGROUND OF THE INVENTION Field of the Invention

[001] The present invention relates to oil-in-water emulsions comprising an oil consisting of one or more polyunsaturated fatty acids, and processes for preparing freezable oil-in-water emulsions. Background

[002] Polyunsaturated fatty acids (PUFAs, including long-chain PUFAs LC-PUFAs) have been shown to improve cognitive function and maintain cardiovascular health, among other benefits. Specifically, omega-3 PUFAs are important components of the diet for preventing arteriosclerosis and coronary heart disease, for alleviating inflammatory conditions, and for slowing the growth of tumor cells, and omega-6 PUFAs are important both as structural lipids and as precursors, for example, to prostaglandins and leukotrienes. PUFAs are an important component of a healthy diet, but since PUFAs are not synthesized by humans in vivo, these compounds must be ingested. For example, plant and animal oils (e.g., fish, nuts, blackberries, hemp, algae, and the seeds and / or leaves of various plants such as flax, chia, perilla, and purslane) are rich in PUFAs.Many people also choose to ingest PUFAs through dietary supplements and / or food products enhanced with PUFAs. As a result, consumer demand for products containing PUFAs has increased. Petition 870210100103, dated 10 / 29 / 2021, pp. 13 / 16 2 / 80 recently, and a wide range of products already contain PUFAs.

[003] Oil-in-water emulsions have been used as a vehicle for PUFAs, both as a precursor for the preparation of edible products, and specifically, as a component of a formulated beverage, food, nutraceutical, and pharmaceutical products. However, PUFAs present in an emulsion can become unstable and degradable (e.g., through oxidation and / or proteolytic degradation), and therefore, maintaining the physical and chemical stability of PUFAs in an emulsion is important. Freezing an oil-in-water emulsion containing PUFAs is an attractive means of reducing the oxidative potential of PUFAs during transport and / or storage. Unfortunately, freezing can lead to demulsification and requires long periods of time to thaw before use.

[004] What is needed is an oil-in-water emulsion that remains fluid and is resistant to demulsification and changes in particle size at subfreezing temperatures (i.e., temperatures below 0°C). In addition, what is needed is a freezable oil-in-water emulsion that is resistant to the oxidative degradation of a PUFA contained therein. BRIEF SUMMARY OF THE INVENTION

[005] The present invention provides thermally stable oil-in-water emulsions comprising an oil comprising a polyunsaturated fatty acid (PUFA), water, and a water-soluble emulsifier and stabilizer selected from: a mixture of sodium chloride and a Petition 870210074972, dated 08 / 16 / 2021, page 13 / 96 3 / 80 monosaccharide, a mixture of propylene glycol and a monosaccharide, and glycerol. When the water-soluble stabilizer is selected from a mixture of sodium chloride and a monosaccharide, or a mixture of propylene glycol and a monosaccharide, or a mixture of propylene glycol and a monosaccharide, then the stabilizer may be present at a concentration of about 20% to about 50% by weight of the emulsion. When the water-soluble stabilizer is glycerol, then the water-soluble stabilizer may be present at a concentration of about 20% to about 55% by weight of the emulsion.In some embodiments, the thermal stability of the oil-in-water emulsion remains at least partially liquefied at a temperature of about -80°C and, in some embodiments, -40°C, and preferably, it is free from variation in particle size and organoleptic properties after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to about 0°C, or at a temperature of about -80°C to about -40°C.

[006] In some embodiments, the emulsion has a shelf life of at least 12 months at freezing temperatures, that is, temperatures equal to or lower than 0°C, preferably between -80°C and 0°C, and even more preferably between about -40°C and about -15°C or -40°C and 0°C, or at a temperature of about -80°C and about 40°C.

[007] In some embodiments, the thermal stability of the oil-in-water emulsion is free from variation Petition 870210074972, dated 08 / 16 / 2021, p. 14 / 96 4 / 80 in particle size and organoleptic properties after 10 freeze-thaw cycles.

[008] In some embodiments, the emulsion has a minimum shelf life of 9 or 12 months at freezing temperatures, i.e., temperatures equal to or lower than 0°C, and is also free from variation in particle size and organoleptic properties after 10 freeze-thaw cycles. For example, in some embodiments, the emulsions of the present invention are free from undesirable variation in particle size and organoleptic properties when stored at temperatures between -40°C and 0°C, or at a temperature of about -40°C to about -15°C, or preferably when stored at temperatures between -80°C and 0°C, or at a temperature of about -80°C to about -40°C.

[009] In some embodiments, sodium chloride is present at a concentration of about 10% to about 25% by weight of the emulsion and a monosaccharide is present at a concentration of about 3% to about 15% by weight of the emulsion. In some embodiments, propylene glycol is present at a concentration of about 10% to about 30% by weight of the emulsion, and a monosaccharide is present at a concentration of about 10% to about 30% by weight of the emulsion. In some embodiments, glycerol is present at a concentration of about 25% to about 55% by weight of the emulsion, for example, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, etc.

[0010] In some embodiments, a PUFA is selected from: α-linolenic acid, γ-linolenic acid, Petition 870210074972, dated 08 / 16 / 2021, p. 15 / 96 5 / 80 linoleic acid, conjugated linoleic acid, arachidonic acid, ω-3 docosapentaenoic acid, ω-3 eicosapentaenoic acid (eicosapentaenoic acid), docosahexaenoic acid and combinations thereof. In some embodiments, PUFA is present at a concentration of about 5% to about 40% by weight of the emulsion.

[0011] In some embodiments, an emulsifying agent is chosen from acacia gum, modified acacia gum, a lecithin, an agar, ghatti gum, modified ghatti gum, pectin, carrageenan, xanthan gum, a modified starch, in particular a modified food starch, a modified alginate, a polyoxyethylene sorbitan, a polyoxyethylene sorbitan ester, a sugar ester, a fatty alcohol, a natural plant product (e.g., quillaja), mono- and / or diglycerides, proteins, and combinations thereof. In some embodiments, an emulsifying agent is present in a concentration of about 10% and about 30% by weight of the emulsion.

[0012] In some embodiments, water is present in a concentration of about 20% to about 60% by weight of the emulsion. In some embodiments, the emulsion has a pH of about 2 to about 7.

[0013] Thus, in one embodiment the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil comprising a polyunsaturated fatty acid, in a concentration of about 5% to about 40% by weight, water, in a concentration of about 20% to about 60% by weight, modified acacia gum or a starch, especially modified starch, such as starch Petition 870210074972, dated 08 / 16 / 2021, p. 16 / 96 6 / 80 food grade modified as an emulsifier and a water-soluble stabilizer which is sodium chloride at a concentration of about 10% to about 25% by weight, and a monosaccharide at a concentration of about 3% to about 15% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of about -40°C, and is free from a change in particle size after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0014] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil comprising a polyunsaturated fatty acid, in a concentration of about 5% to about 40% by weight, water, in a concentration of about 20% to about 60% by weight, modified acacia gum or a starch, especially a modified starch, such as a starch modified as an emulsifier and a water-soluble stabilizer, which is propylene glycol, in a concentration of about 10% to about 30% by weight, and a monosaccharide with a concentration of about 10% to about 30% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of about -40°C, and is free from a change in particle size after 9 months of storage at a temperature of about -40°C to about 0°C.or at a temperature of about -40°C to about 15°C or at a temperature of about -80°C to about Petition 870210074972, dated 16 / 08 / 2021, page 17 / 96, 7 / 80 40°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0015] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil consisting of a polyunsaturated fatty acid at a concentration of about 5% to about 40% by weight, water at a concentration of about 20% to about 60% by weight, modified acacia gum or a starch, especially a modified starch, such as a modified starch as an emulsifier, and a water-soluble stabilizer, which is glycerol, at a concentration of about 30% to about 40% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of about 40°C, and is free from variation in particle size and organoleptic properties after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C,or at a temperature of about -80°C to about -40°C.

[0016] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil comprising a polyunsaturated fatty acid, in a concentration of about 5% to about 40% by weight, of Quillaia saponaria as an emulsifier, and a water-soluble stabilizer which is glycerol, in a concentration of about 20% to about 40% by weight, and the other components as desired, in which the thermally stable oil-in-water emulsion remains at least partially liquefied at a Petition 870210074972, dated 08 / 16 / 2021, p. 18 / 96 8 / 80 temperature of -40°C, or at a temperature of about 80°C to 0°C, or at a temperature of about -80°C to about -40°C and is free from a variation in particle size and organoleptic properties after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0017] In some embodiments, an oil-in-water emulsion of the present invention is substantially free of saccharides.

[0018] In some embodiments, an emulsion further comprises a flavor-masking agent. In some embodiments, an emulsion further comprises a preservative.

[0019] In some embodiments, an emulsion of the present invention further comprises an antioxidant. Antioxidants suitable for use with the present invention include, but are not limited to, vitamin C, vitamin E, a polyphenol, a phenol derivative, carnosic acid, lipoic acid, taurine, an aromatic carboxylic acid, salts of an aromatic carboxylic acid, amino acids that have antioxidant properties, proteins that have antioxidant properties, and combinations thereof.

[0020] The present invention is also directed to processes for preparing a thermally stable oil-in-water emulsion. In some embodiments, a process comprises: A combination of water and an emulsifier to provide an aqueous mixture. Petition 870210074972, dated 08 / 16 / 2021, p. 19 / 96 9 / 80 addition to the aqueous mixture of an oil comprising a polyunsaturated fatty acid, while mixing to provide an oil-in-water emulsion, and addition to the oil-in-water emulsion of a water-soluble stabilizer selected from: a mixture of sodium chloride and a monosaccharide, a mixture of propylene glycol and a monosaccharide, and glycerol, wherein the water-soluble stabilizer is present in a concentration of about 20% to about 50% by weight of the emulsion to provide a thermally stable oil-in-water emulsion, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of about -40°C and is free from a variation in particle size after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C at 0°C, or at a temperature of about -80°C to about -40°C. Other embodiments, features and advantages of the present invention, as well as the structure and operation of the various embodiments of the present invention, are described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE FIGURES

[0021] The accompanying drawings, which are incorporated into this document and form part of the descriptive report, illustrate one or more embodiments of the present invention and, together with the description, further serve to explain the principles of the invention and to enable one skilled in the relevant art to make use of the invention. Petition 870210074972, dated 08 / 16 / 2021, p. 20 / 96 10 / 80

[0022] Figure 1 shows the average particle size (μm) of the emulsion from Example 4, Table 5, before one freeze / thaw cycle (square), after 1 freeze / thaw cycle (triangle), after 2 freeze / thaw cycles (x), and after three freeze / thaw cycles (star).

[0023] Figure 2 shows a processing diagram for obtaining a thermally stable (freezable) emulsion.

[0024] One or more embodiments of the present invention will now be described with reference to the accompanying drawings. DETAILED DESCRIPTION OF THE INVENTION

[0025] This specification describes one or more embodiments that incorporate the features of the present invention. The embodiments described merely exemplify the invention. The scope of the invention is not limited to the embodiment(s) described. The invention is defined by the appended claims.

[0026] Throughout this disclosure, all expressions of percentage, ratio, incorporation and the like are by weight unless otherwise indicated. As used herein, by weight is synonymous with the term by mass and indicates that a proportion or percentage defined herein is calculated according to weight rather than volume, or some other measure.

[0027] As used in this document, composition and mixture are used interchangeably and refer to a combination of two or more materials, substances, excipients, parts and the like. Petition 870210074972, dated 08 / 16 / 2021, page 21 / 96 11 / 80

[0028] As used in this document, homogeneous refers to mixtures, compositions, and specifically emulsions with a substantially uniform distribution, for example, of oil particles in a continuous aqueous phase. Homogeneity is synonymous with uniformity and may refer to intrasample uniformity, batch-to-batch uniformity, and / or operation-to-operation uniformity. For example, intrasample uniformity can be determined by analyzing a first portion of an emulsion, mixture, or composition and comparing it with a second portion of the same emulsion, mixture, or composition. Typical deviations of a composition (e.g., variation in the weight percentage of excipients, particle size, and the like) from a substantially homogeneous mixture are about 10% or less, about 5% or less, about 3% or less, about 2% or less, about 1% or less, or within experimental error.

[0029] The oil-in-water emulsions of the present invention comprise an immiscible mixture of a continuous aqueous liquid phase and a discontinuous oil phase. As used herein, continuous aqueous liquid phase refers to the portion of the emulsion in which the discontinuous oil phase is dispersed. Consequently, a discontinuous oil phase refers to the multiplicity of separate elements dispersed within, and immiscible with, the continuous aqueous liquid phase. The discontinuous oil phase is present in the form of particles. As used herein, particles refers to an oil phase of an emulsion comprising a plurality of Petition 870210074972, dated 08 / 16 / 2021, p. 22 / 96 12 / 80 separate particles. As used herein, particle size refers to particle diameter, which is the diameter of the base particles in an approximate spherical shape based on a volumetric measurement of the particle. In addition to spherical particles, the oil-in-water emulsions of the present invention may also comprise, without limitation, hemispherical, ellipsoidal and / or cylindrical particles.

[0030] Assessing whether there has been a change in the particle size distribution of the emulsion over time is a good measure of the emulsion's stability. The absence of change, or a small change in the particle size distribution of the emulsion over time, indicates that the emulsion is stable. As used herein, particle size distribution refers to the number or concentration (e.g., percentage) of particles that have a given size (i.e., diameter), or the range of sizes, within a given batch of emulsion and / or batch of the invention. As used herein, a particle size distribution refers to the number or concentration (e.g., percentage) of particles that have a given size (i.e., diameter), or the range of sizes, within a given batch of emulsion and / or batch of the invention.Particle size and particle size distribution can be measured using Low Angle Laser Scattering (LALLS), for example with a Mastersizer Hydro 2000S (Malvern Instruments Ltd., Worcestershire, UK). Particle size and particle size distribution can also be... Petition 870210074972, dated 08 / 16 / 2021, page 23 / 96 13 / 80 measured by, for example, microphotography, video microscopy, video-enhanced microscopy, Coulter counting, differential scanning calorimetry, turbidimetry, dynamic and / or static light scattering, low-intensity ultrasound, nuclear magnetic resonance, or any other particle measurement technique of known size to those skilled in the common art.

[0031] As used in this document, a D50 value, or d(0.5), refers to the particle size of an oil phase and, specifically, the diameter at which 50% of the measurable particles of the oil phase have a larger equivalent diameter, and the other 50% of the particles have a smaller equivalent diameter. So, D50 generally refers to the average particle diameter.

[0032] In some embodiments, the discontinuous oil phase particles have a weighted (average) particle size of about 20 nm to about 1.5 μm, about 50 nm to about 1 μm, about 100 nm to about 1.5 μm, preferably about 100 nm to about 1 μm, about 150 nm to about 700 nm, or about 200 nm to 500 nm.

[0033] As used in this document, a D90 value, or d(0.9), refers to the particle size of an oil phase and, specifically, the diameter at which 90% of all measurable particles in the oil phase have a diameter equal to or less than the D90 value, and 10% of the measurable particles will have a diameter greater than the D90 value. Petition 870210074972, dated 08 / 16 / 2021, p. 24 / 96 14 / 80

[0034] In some embodiments, the discontinuous oil phase particles have a D90 of about 10 μm or less, about 5 μm or less, about 2 μm or less, or about 1 μm or less.

[0035] As used in this document, a D10 value, or d(0,1), refers to the particle size of an oil phase and, specifically, the diameter at which 10% of all measurable particles in the oil phase have a diameter equal to or less than the D10 value, and 90% of the measurable particles have a diameter greater than the D10 value.

[0036] In some embodiments, the discontinuous oil phase particles have a D10 of about 50 nm or less, about 60 nm or less, about 70 nm or less, about 80 nm or less, about 90 nm or less, about 100 nm or less, about 200 nm or less, about 250 nm or less, about 300 nm or less, about 400 nm or less, or about 500 nm or less.

[0037] As used in this document, a D100 value or d(1.ooo) refers to the particle size of an oil phase and, specifically, the diameter at which 100% of all measurable particles in the oil phase have a diameter equal to or less than the D100 value, and 0% of the measurable particles have a diameter greater than the D100 value.

[0038] The particle size distribution in a mixture can also be defined by the ratio D10:D50, the ratio D10:D90, and the ratio D50:D90. In some embodiments, the particle size distribution of an oil-in-water emulsion of the present invention is such that the distribution Petition 870210074972, dated 08 / 16 / 2021, p. 25 / 96 15 / 80 of the emulsion has a Dio:Doo ratio of about 1:10 or less, about 1:8 or less, about 1:6 or less, about 1:5 or less, or about 1:3 or less. In some embodiments, the particle size distribution in a mixture, or emulsion, may also be defined by the set of particles that are between about 0.1 μm and about 0.36 μm in diameter. In some embodiments, the percentage of particles that are in the range of about 0.01 μm to about 0.36 μm in diameter is greater than about 95%, greater than about 96%, greater than about 97%, greater than about 98%, greater than about 99%, or 100% of the particles.

[0039] In some embodiments, it is preferred that at least 90% of the particles have a particle size between about 0.02 and about 0.36 microns.

[0040] As employed in this document, a substantial variation in particle size refers to an increase in any of D10, D50 and / or D90 of about 10% or more, for example, about 20% or more, about 25% or more, about 30% or more, or about 40% or more. In some embodiments, an oil-in-water emulsion of the present invention can be stored for a period of 9 months or more, or a year or more, without substantial variation in particle size.

[0041] As used in this document, D [3,2] refers to the particle size of an oil phase and, specifically, to the surface weighted average diameter.

[0042] As used in this document, D[4,3] refers to the particle size of a phase of Petition 870210074972, dated 08 / 16 / 2021, p. 26 / 96 16 / 80 oil, and specifically to the volume-weighted average diameter.

[0043] As used in this document, emulsion stability refers to the ability of an emulsion to resist changes in the physical and chemical properties of the emulsion, including physical destabilization such as creaming, flocculation, coalescence, partial coalescence, phase inversion and Ostwald ripening over time, and chemical changes in the emulsion formulation to protect and stabilize a PUFA, for example, from oxidation. Changes in physical instability are reflected in a change in one or more physical properties of the emulsion and may include, for example, a change in pH, viscosity, particle size and / or distribution.

[0044] As used in this document, uniformity refers to absolute deviations from the mean.

[0045] As used in this document, unless otherwise indicated or evident from the context, the terms or less or less than refer to percentages that include 0% or quantities not detectable by ordinary means.

[0046] The thermally stable oil-in-water emulsions of the present invention comprise an oil consisting of one or more polyunsaturated fatty acids (PUFAs), water, an emulsifier and a water-soluble stabilizer selected from: (1) a mixture of sodium chloride and a monosaccharide, (2) a mixture of propylene glycol and a monosaccharide, and (3) glycerol. When the water-soluble stabilizer is selected from a mixture of Petition 870210074972, dated 08 / 16 / 2021, page 27 / 96 17 / 80 sodium chloride and a monosaccharide, or a mixture of propylene glycol and a monosaccharide, then the stabilizer may be present at a concentration of about 20% to about 50% by weight of the emulsion. When the water-soluble stabilizer is glycerol, then the water-soluble stabilizer may be present at a concentration of about 20% to about 55% by weight of the emulsion. The thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of about 40°C, and preferably is free from variation in particle size and organoleptic properties after 9 months of storage at a temperature of about -40°C to about -15°C, or at a temperature of about 40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about 40°C.

[0047] The thermally stable oil-in-water emulsions of the present invention are particularly advantageous because the emulsions can be stored at a temperature below 0°C for an extended period of time (e.g., 6 months or more, 9 months or more, or a year or more) without undergoing a significant change in particle size or organoleptic quality.

[0048] For example, in some embodiments an oil-in-water emulsion of the present invention is free from particle size variation when stored at about -40°C to about -15°C for a period of 9 months or more, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, Petition 870210074972, dated 08 / 16 / 2021, p. 28 / 96 18 / 80 or at a temperature of about -80°C to about -40°C for a period of 9 months or more.

[0049] As employed in this document, a variation in particle size may refer to an increase in any of D10, D50 and / or D90 of about 40% or more, about 30% or more, about 25% or more, about 20% or more, or about 10% or more. Alternatively, a variation in particle size may also refer to a decrease in the percentage of particles that are larger than a specific size range. In some embodiments, the decrease in the percentage of particles that fall within a specific size range is less than 95%. In some embodiments, the specific size range is from about 0.01 to about 0.36 μm. In some embodiments, an oil-in-water emulsion of the present invention may be stored for a period of 9 months or more, or one year or more, without variation in particle size or particle size distribution.

[0050] In some embodiments, thermally stable oil-in-water emulsions are substantially free from coalescence, partial coalescence, flocculation, Ostwald ripening, creaming, sedimentation, demulsification, phase inversion, changes in particle size, and / or changes in organoleptic properties and the like, after six months or more at a temperature of about -40°C to about -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C. Petition 870210074972, dated 08 / 16 / 2021, p. 29 / 96 19 / 80

[0051] The thermally stable oil-in-water emulsions of the present invention are freezable and can be formulated to resist freezing down to a temperature of -40°C, or, in some embodiments, to a temperature of about -80°C. As used herein, when referring to the oil-in-water emulsions of the present invention as being freezable, it is intended that the oil-in-water emulsions remain in a fluid, non-solid state at a temperature below the freezing point of water, for example, at temperatures as low as about -41°C. In some embodiments, the emulsion remains fluid between about -40°C and about 15°C, at a temperature of about -40°C and about 0°C, or between -80°C and 0°C, or at temperatures between -80°C and 40°C.

[0052] In some embodiments, the oil-in-water emulsions remain in a fluid, non-solid state at freezing temperatures, for example, between about 80°C and 0°C, and preferably at temperatures between -80°C, 75°C, -70°C, -65°C, -60°C, -55°C, -50°C, -45°C, -40°C, 35°C, -30°C, -25°C, -20°C, -15°C, -10°C, -5°C, 0°C, 5°C, 10°C, or 5°C. The thermally stable oil-in-water emulsions of the present invention are fluid, that is, they remain fluid, at temperatures at which refrigerated or ambient emulsions may freeze, for example, at about -17°C to about -21°C. Thus, they are easy to use since they can be employed in many applications without thawing before use. For example, oil-in-water emulsions can be removed from frozen storage (i.e., storage at a Petition 870210074972, dated 08 / 16 / 2021, p. 30 / 96 20 / 80 temperature at which water would normally freeze), that is, about -40°C, for example, and used immediately without thawing (or liquefying) the oil-in-water emulsions. If desired, the emulsion temperature can be increased to room temperature or higher before use after removal from frozen storage.

[0053] As used in this document, the term flowable refers to the ability of a composition to be transported by gravity or by mechanical processes or by pneumatic or conventional pumping from a storage container. Thus, from the point of view of ease of use and cost of ownership, the thermally stable oil-in-water emulsions of the present invention provide a significant advantage over emulsions that solidify at temperatures equal to or lower than 0°C.

[0054] As employed in this document, a freeze-thaw cycle refers to a process whereby an oil-in-water emulsion is cooled to a temperature of about -40°C to 0°C (or to a different temperature if so stated), held at this temperature for at least 24 hours, and then returned to ambient temperature (e.g., 4°C to 25°C). In some embodiments, the thermally stable oil-in-water emulsions of the present invention are free from particle size variation during at least five or six freeze-thaw cycles, and preferably during at least 12 freeze-thaw cycles. Petition 870210074972, dated 08 / 16 / 2021, p. 31 / 96 21 / 80 a change in particle size after 10 or more freeze-thaw cycles, 12 or more, 15 or more, 20 or more, 25 or more, or 30 or more freeze-thaw cycles.

[0055] A specific advantage is that the oil-in-water emulsions of the present invention are substantially liquid at a temperature of about -40°C to -15°C or at a temperature of about -40°C to about 0°C or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C. Thus, while some solids may form in an oil-in-water emulsion of the present invention, the oil-in-water emulsion remains in a non-solid state even when subjected to freezing temperatures and, especially, temperatures as low as -40°C and, especially in some embodiments, about -80°C.

[0056] Not being tied to any specific theory, as an oil-in-water emulsion of the present invention is cooled below 0°C, which is the normal freezing temperature of water, the formation of aqueous solids is inhibited by the presence of a water-soluble stabilizer, which is present at a concentration of 20% to 55% by weight of the emulsion. Thus, the thermally stable oil-in-water emulsions of the present invention remain substantially liquefied at a temperature of -40°C, or at a temperature of about -80°C.

[0057] In some embodiments, the present invention provides a thermally stable oil-in-water emulsion consisting of an oil comprising a polyunsaturated fatty acid oil, water, an emulsifier and a Petition 870210074972, dated 08 / 16 / 2021, p. 32 / 96 22 / 80 Water-soluble stabilizer selected from: a mixture of sodium chloride and a monosaccharide, a mixture of propylene glycol and a monosaccharide, or glycerol. When the water-soluble stabilizer is selected from a mixture of sodium chloride and a monosaccharide, or a mixture of propylene glycol and a monosaccharide, then the stabilizer may be present at a concentration of about 20% to about 50% by weight of the emulsion. When the water-soluble stabilizer is glycerol, then the water-soluble stabilizer may be present at a concentration of about 20% to about 55% by weight of the emulsion.The thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of -40°C and preferably is free from variation in particle size and organoleptic properties after 9 months of storage at a temperature of -40°C to -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0058] In some embodiments, the emulsion has a minimum shelf life of 9 months or 12 months at freezing temperatures, i.e., temperatures equal to or below 0°C, for example, in some embodiments, the emulsion has a minimum shelf life of 9 months or 12 months at -40°C to -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about 80°C to 0°C, or at a temperature of about -80°C to about -40°C. In some embodiments, the thermally stable oil-in-water emulsion is free from variation in size of Petition 870210074972, dated 08 / 16 / 2021, p. 33 / 96 23 / 80 particles and in organoleptic properties after 10 freeze-thaw cycles. In some embodiments, the emulsion has a minimum shelf life of 9 or 12 months at freezing temperatures, i.e., temperatures equal to or below 0°C, and is also free from variation in particle size and organoleptic properties after 10 freeze-thaw cycles.

[0059] In some embodiments, sodium chloride is present at a concentration of about 10% to about 25% by weight of the emulsion, and a monosaccharide is present at a concentration of about 3% to about 15% by weight of the emulsion. In some embodiments, propylene glycol is present at a concentration of about 10% to about 30% by weight of the emulsion, and a monosaccharide is present at a concentration of about 10% to about 30% by weight of the emulsion. In some embodiments, glycerol is present at a concentration of about 25% to about 55% by weight of the emulsion, for example, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, etc.

[0060] In some embodiments, a PUFA is selected from: α-linolenic acid, γ-linolenic acid, linoleic acid, conjugated linoleic acid, arachidonic acid, ω-3 docosapentaenoic acid, ω-6 docosapentaenoic acid, eicosapentaenoic acid, docosahexaenoic acid, and combinations thereof. In some embodiments, the PUFA is present at a concentration of about 5% to about 40% by weight of the emulsion. In some embodiments, a PUFA is present in the oil-in-water emulsion in an amount of about 50 mg to about 80 mg per gram of emulsion, about 60 mg Petition 870210074972, dated 08 / 16 / 2021, p. 34 / 96 24 / 80 at approximately 75 mg per gram of emulsion, or from approximately 65 mg to approximately 70 mg per gram of emulsion.

[0061] In some embodiments, an emulsifying agent is chosen from among: modified acacia gum, lecithin, agar, modified ghatti gum, pectin, carrageenan, xanthan gum, modified food starch, modified alginate, a polyoxyethylene sorbitan ester, sugar ester and combinations thereof. In some embodiments, an emulsifying agent is present in a concentration of about 10% to about 30% by weight of the emulsion.

[0062] In some forms, water is present in a concentration of about 20% to about 60% by weight of the composition.

[0063] Thus, in one embodiment the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil comprising a polyunsaturated fatty acid, in a concentration of about 5% to about 40% by weight, water, in a concentration of about 20% to about 60% by weight, an emulsifier such as, for example, modified acacia gum and / or a starch, especially a modified starch, such as a modified food starch, for example, quillaja saponaria, and a water-soluble stabilizer which is sodium chloride in a concentration of about 10% to about 25% by weight and a monosaccharide, in a concentration of about 3% to about 15% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of -40°C, and is free from a change in particle size after 9 months of Petition 870210074972, dated 08 / 16 / 2021, p. 35 / 96 25 / 80 storage at a temperature of -40°C to -15°C, or at a temperature of about -40°C to about 0°C, or at a temperature of about -80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0064] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil consisting of a polyunsaturated fatty acid at a concentration of about 5% to about 40% by weight, water at a concentration of about 20% to about 60% by weight, an emulsifier such as, for example, modified acacia gum and / or a starch, especially a modified starch, such as a modified food starch, for example, quillaja saponaria, and a water-soluble stabilizer which is propylene glycol at a concentration of about 10% to about 30% by weight and a monosaccharide at a concentration of about 10% to about 30% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of -40°C, or at a temperature of about 80°C. at 0°C, or at a temperature of about -80°C to about -40°C,and is free from particle size variation after 9 months of storage at a temperature of 40°C to -15°C, or at a temperature of approximately -40°C to approximately 0°C, or at a temperature of approximately -80°C to 0°C, or at a temperature of approximately -80°C to approximately -40°C.

[0065] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil consisting of a polyunsaturated fatty acid at a concentration of about 5% to Petition 870210074972, dated 08 / 16 / 2021, p. 36 / 96 26 / 80 approximately 40% by weight of water, at a concentration of approximately 20% to approximately 60% by weight, an emulsifier such as, for example, modified acacia gum and / or a starch, especially a modified starch such as a modified food starch, for example, quillaja saponaria, and a water-soluble stabilizer which is glycerol, at a concentration of approximately 30% to approximately 40% by weight, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of 40°C, and is free from variation in particle size and organoleptic properties after 9 months of storage at a temperature of -40°C to -15°C, or at a temperature of approximately -40°C to approximately 0°C, or at a temperature of approximately -80°C to approximately 0°C, or at a temperature of approximately -80°C to approximately 0°C. -40°C.

[0066] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil consisting of polyunsaturated fatty acid, at a concentration of about 5% to about 40% by weight, and a starch, especially a modified starch such as a modified food starch, for example, quillaja saponaria as an emulsifier, and a water-soluble stabilizer, which is chosen from: (1) a mixture of sodium chloride and a monosaccharide, (2) a mixture of propylene glycol and a monosaccharide and (3) glycerol, such stabilizer being present at a concentration of 2940% by weight, and the other components as desired, wherein the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of -40°C or -80°C, and is free from a Petition 870210074972, dated 08 / 16 / 2021, p. 37 / 96 27 / 80 variation in particle size and organoleptic properties after 9 months of storage at a temperature of -40°C to -15°C or at a temperature of about -40°C to about 0°C, or at a temperature of about 80°C to 0°C, or at a temperature of about -80°C to about -40°C.

[0067] In another embodiment, the present invention is directed to a thermally stable oil-in-water emulsion comprising: an oil consisting of a polyunsaturated fatty acid, at a concentration of about 5% to about 20% by weight, about 10% to about 40% of a polymeric hydrocolloid from a plant source, and a water-soluble stabilizer which is glycerol, at a concentration of about 5% to about 45% by weight, about 15% to about 50% by weight of water, and other components as desired.

[0068] The individual components of the emulsifiers of the invention claims are further described below. Water-soluble stabilizer

[0069] The thermally stable emulsions of the present invention comprise a water-soluble stabilizer selected from: a mixture of sodium chloride and a monosaccharide, a mixture of propylene glycol and a monosaccharide, and glycerol. In addition to preventing the solidification of an oil-in-water emulsion of the present invention, the water-soluble stabilizer can allow the formation of small particles of the oil phase and stabilize the particle size of the discontinuous oil phase through multiple freeze-thaw cycles. Petition 870210074972, dated 08 / 16 / 2021, page 38 / 96 28 / 80 thawing. In some embodiments, the water-soluble stabilizer can also improve one or more organoleptic properties (e.g., taste, aroma, texture, and the like) of oil-in-water emulsions. In some embodiments, the products containing the emulsions of the present invention have good organoleptic properties compared to products that do not contain the emulsion. In some embodiments, there is no difference or very little / trace difference between the products that have the emulsion and the products that do not have the emulsion.

[0070] In some embodiments, the water-soluble stabilizer reduces the viscosity and / or surface tension of the continuous liquid aqueous phase. As a result, the discontinuous oil phase, which includes a PUFA, can be more easily dispersed in the oil-in-water emulsion. In some embodiments, the stability of the oil-in-water emulsions can also be improved by minimizing the density difference between the discontinuous oil phase and the continuous aqueous phase.

[0071] Water-soluble stabilizers suitable for use with the oil-in-water emulsions of the present invention include a mixture of sodium chloride and a monosaccharide, a mixture of propylene glycol and a monosaccharide, or glycerol.

[0072] The total concentration of the water-soluble stabilizing agent in an oil-in-water emulsion of the present invention is from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 25%, from about 35% to about 40%, from about 20%, from about 25%, from about 35%, or from about 37% by weight. Petition 870210074972, dated 08 / 16 / 2021, p. 39 / 96 29 / 80

[0073] In some embodiments, sodium chloride is present at a concentration of about 10% to about 25% by weight of an emulsion. In some embodiments, a monosaccharide is present at a concentration of about % to about 15% by weight of an emulsion. In some embodiments, a monosaccharide is present at a concentration of about 5% to about 10% by weight of an emulsion. In some embodiments, sodium chloride and a monosaccharide are present in a ratio of about 1:1.5 to about 5:1, 1:1 to 4:1, 1:1 to 3:1, 2:1 to 3:1, 1:1, 2:1, 2.5:1, 3:1, or 4:1.

[0074] In some embodiments, propylene glycol is present at a concentration of about 10% to about 30% by weight of the emulsion, and a monosaccharide is present at a concentration of about 10% to about 30% by weight of the emulsion. In some embodiments, propylene glycol and a monosaccharide are present in a ratio of about 3:5 to about 5:3, 3:4 to 4:3, or 1:1.

[0075] In some embodiments, glycerol (glycerin) is present in a concentration of about 46 to about 55%, about 40 to about 46%, about 25% to about 40%, about 30% to about 40%, about 32% to about 38%, about 34% to about 38%, or about 36% by weight of an emulsion. In some embodiments, glycerol may be present in a concentration greater than about 45%, for example, about 46%, about 47%, about 48%, about 49%, about 50%, about 51%, about 52%, about 53%, about 54%, or about 55%. The amount of glycerol can be greater than 45%, as above, provided that sufficient water is present to hydrate the... Petition 870210074972, dated 08 / 16 / 2021, p. 40 / 96 30 / 80 emulsifier so that the emulsifying starch will facilitate encapsulation to a sufficient degree.

[0076] Changes in the pH of the emulsion over time are a measure of the emulsion's stability. In a preferred embodiment, the emulsion formulation is well buffered. Such buffering improves the ability of the emulsion formulation to withstand changes in the process or realization and use, changes in its ingredients, and other factors.

[0077] In some embodiments, an oil-in-water emulsion of the present invention is acidic, and in some embodiments, it may be pH neutral or basic. When potassium sulfate is used as a preservative, it is preferable that the oil-in-water emulsion be acidic. The pH of an oil-in-water emulsion can be controlled by adding an appropriate amount of an acid and / or a base. Suitable acids and bases for use with the present invention include, but are not limited to, acetic acid, citric acid, hydrochloric acid, sodium hydroxide, sodium carbonate, sodium bicarbonate, and the like. Not limited by any particular theory, an acidic pH can stabilize the oil-in-water emulsion during storage. In some embodiments, anhydrous citric acid is present in an amount of about 3% to about 8% by weight of an emulsion.

[0078] In some embodiments, an oil-in-water emulsion of the present invention has a pH of about 7 or less, about 6 or less, about 5 or less, or about 4 or less. In some embodiments, an oil-in-water emulsion has a pH of about 2 to about 7, about 2.5 Petition 870210074972, dated 08 / 16 / 2021, p. 41 / 96 31 / 80 to about 6, about 3 to about 5, about 3.5 to about 5, about 4 to about 4.5, or about 4. Aqueous Liquid Phase

[0079] The continuous aqueous liquid phase includes an aqueous liquid, for example, a beverage, that is compatible with a PUFA oil, an emulsifier, and a water-soluble stabilizer. Suitable aqueous liquids for use with the continuous aqueous liquid phase include, but are not limited to, water, carbonated water, syrup, diet drinks, soft drinks, fruit juices (including, but not limited to, green grape, dark grape, mixed berries, tropical blends, orange / pineapple / mango, strawberry / banana, pomegranate / blackberry, green grape / raspberry), vegetable juices, isotonic drinks, non-isotonic drinks, soft drinks containing fruit juice, coffee, tea, milk and dairy products (e.g., milk, cream, and similar), soy products (e.g., milk) and similar products, and combinations thereof.

[0080] In some embodiments, an aqueous liquid component (e.g., water) is present in a concentration of about 20% to about 75%, about 20% to about 60%, about 25% to about 65%, about 25% to about 60%, about 25% to about 50%, about 25% to about 45%, about 28% to about 35%, about 35% to about 50%, about 40% to about 50%, about 40% to about 45%, about 28%, about 30%, about 35%, about 40%, or about 45% by weight of an emulsion.

[0081] In some embodiments, a continuous aqueous liquid phase (i.e., the aqueous liquid and any excipients soluble therein) comprises about 55% to about 95%, about 60% to about 95%, about 70% to about Petition 870210074972, dated 08 / 16 / 2021, p. 42 / 96 32 / 80 of 90%, about 80% to about 90%, about 80% to about 90%, about 80%, about 85%, or about 90% by weight of an oil-in-water emulsion. In some embodiments, a discontinuous oil phase is present in an oil-in-water emulsion of the present invention at a concentration of about 5% to about 45%, about 5% to about 40%, about 10% to about 30%, about 10% to about 20%, about 15% to about 30%, about 15% to about 25%, about 15%, or about 20% by weight of the emulsion. PUFA Oils

[0082] The thermally stable emulsions of the present invention comprise a PUFA oil. The thermally stable emulsions of the invention provide an emulsion that can be used to deliver safe and effective administration of a PUFA oil.

[0083] The oil-in-water emulsions of the present invention provide a safe and effective administration of a PUFA oil. As used herein, a PUFA (PUFA) refers to a fatty acid having a backbone comprising 16 or more carbon atoms (e.g., 16, 18, 20, or 22 carbon atoms (C16, C18, C20, or C22, respectively)), and two or more carbon-carbon double bonds in the structure. As used herein, a long-chain PUFA (LC-PUFA) refers to a fatty acid having a backbone comprising 18 or more carbon atoms, and two or more carbon-carbon double bonds in the backbone, for example, C18:3n3 (alpha-linolenic acid or ALA). When the notation CA:Bn-X is used for a methylene-interrupted PUFA, CA is the number of atoms of Petition 870210074972, dated 08 / 16 / 2021, p. 43 / 96 33 / 80 carbon (e.g., C18, C20, or C22), B is the number of double bonds, and X is the position of the first double bond from the methyl end of the fatty acid chain.

[0084] As used herein, the term PUFA encompasses the free acid form thereof, as well as their salts and esters. As used herein, the term ester refers to the substitution of the hydrogen in the carboxylic acid group of a PUFA molecule with another substituent. Typical esters are known to those skilled in the art, a discussion of which is provided by Higuchi, T. et al., Pro-drugs as Novel Delivery Systems, vol. 14, ACS Symposium Series, Bioreversible Carriers in Drug Design, ed. Edward B. Roche, Amer. Pharma. Assoc., Pergamon Press (1987), and Protective Groups in Organic Chemistry, McOmie ed., Plenum Press, New York (1973), each of which is hereby incorporated by reference in its entirety. Examples of common esters include methyl, ethyl, trichloroethyl, propyl, butyl, pentyl, t-butyl, benzyl, nitrobenzyl, methoxybenzyl, and benzhydryl.Other PUFA esters are described in U.S. Patent Application Publication Number 2010-0130608 A1, which is incorporated herein by reference in its entirety.

[0085] PUFAs for use with the present invention include omega-3, omega-6 and omega-9 fatty acids, and oxylipins derived therefrom. Exemplary omega-3 PUFAs for use with the present invention include, but are not limited to, α-linolenic acid (C18:3n-3), C18:4n-4, ω-3 eicosapentaenoic acid (20:5n-3), ω-3 docosapentaenoic acid (docosapentaenoic acid), ω-3 docosahexaenoic acid (22:6n-3), Petition 870210074972, dated 08 / 16 / 2021, p. 44 / 96 34 / 80 docosatetraenoic acid (22:4n-6) and its combinations. Exemplary Omega-6 PUFAs for use with the present invention include, but are not limited to, γ-linolenic acid, linoleic acid, conjugated linoleic acid, arachidonic acid (20:4n-6), docosapentaenoic acid-6 and their combinations. In some embodiments, a PUFA oil for use with the present invention is all-cis acid.

[0086] In some embodiments, PUFA comprises DHA. DHA refers to docosahexaenoic acid, also known by its chemical name (all-Z) 4,7,10,13,16,19-docosahexaenoic acid, as well as any salts or derivatives thereof. Thus, the term DHA encompasses DHA ethyl ester (DHA-EE), as well as DHA-free fatty acids, phospholipids, other esters, monoglycerides, diglycerides, and triglycerides containing DHA. DHA is a polyunsaturated ω-3 fatty acid.

[0087] The term ester in the term ethyl ester-DHA refers to the substitution of the hydrogen of the carboxylic acid group of the DHA molecule by an ethyl group. In some embodiments, the ester substituent can be added to the free DHA acid molecule when the DHA is in a purified or semi-purified state. Alternatively, the DHA ester is formed after the conversion of a triglyceride into an ester.

[0088] In some embodiments, the PUFA oil used to make the emulsion thermally stable is substantially free of one or more specific fatty acids. For example, a PUFA oil containing DHA-EE may be substantially free of eicosapentaenoic acid (EPA). Petition 870210074972, dated 08 / 16 / 2021, p. 45 / 96 35 / 80

[0089] EPA refers to eicosapentaenoic acid, known by its chemical name (all-Z) 5,8,11,14,17-eicosapentaenoic acid, as well as any salts or derivatives thereof. Thus, the term EPA encompasses free EPA acid as well as alkyl esters of EPA and EPA-containing triglycerides. EPA is a polyunsaturated ω-3 fatty acid. Unless otherwise indicated, an oil that is used to obtain the thermally stable emulsion that is substantially free of EPA refers to an oil in which EPA is less than about 3%, by weight, of the total fatty acid content of the oil.In some embodiments, the oil used to obtain the thermally stable emulsion comprises less than about 2% EPA by weight of the total fatty acid content of the oil, less than about 1% EPA by weight of the total fatty acid content of the oil, less than about 0.5% EPA by weight of the total fatty acid content of the oil, less than about 0.2% EPA by weight of the total fatty acid content of the oil, or less than about 0.01% EPA by weight of the total fatty acid content of the oil. In some embodiments, the oil has no detectable amount of EPA.

[0090] As used in this document, a substantially EPA-free emulsion may refer to an emulsion in which EPA is less than about 3% by weight of the total fatty acid content of the emulsion. In some embodiments, the emulsion includes less than about 2% EPA by weight of the total fatty acid content of the emulsion, less than about 1% EPA by weight of the total fatty acid content of the emulsion, less than about 0.5% EPA by weight of the total fatty acid content of the emulsion, less Petition 870210074972, dated 08 / 16 / 2021, p. 46 / 96 36 / 80 of approximately 0.2% EPA by weight of the total fatty acid content of the emulsion, or less than approximately 0.01% EPA by weight of the total fatty acid content of the emulsion. In some embodiments, the emulsion does not contain any detectable amount of EPA.

[0091] In some embodiments, the oil or emulsion containing DHA, or especially containing DHA-EE, is substantially free of 22:5n-6 docosapentaenoic acid (DPAn6). The term DPAn6 refers to docosapentaenoic acid, an omega-6 fatty acid, known by its chemical name (all-Z)-4,7,10,13,16-docosapentaenoic acid, as well as any salts or esters thereof. Thus, the term DPAn6 encompasses free DPAn6 acid, as well as DPAn6 ethyl esters and DPAn6-containing triglycerides. DPAn6 may be removed during DHA purification, or alternatively, DHA may be obtained from an organism that does not produce DPAn6, or produces very little DPAn6.

[0092] As used in this document, a substantially DPAn6-free oil refers to an oil used to obtain the emulsion that contains less than about 2% by weight of docosapentaenoic acid 22:5n-6 (DPAn6) of the total fatty acid content of the oil. In some embodiments, the oil contains less than about 1% DPAn6 by weight of the total fatty acid content of the oil. In some embodiments, the oil contains less than about 0.5% DPAn6 by weight of the total fatty acid content of the oil. In some embodiments, the oil contains no detectable amount of DPAn6. Petition 870210074972, dated 08 / 16 / 2021, p. 47 / 96 37 / 80

[0093] As used herein, a substantially DPAn6-free emulsion refers to an emulsion containing less than about 2% by weight of docosapentaenoic acid 22:5 n-6 (DPAn6) of the total fatty acid content of the emulsion. In some embodiments, the emulsion contains less than about 1% DP n6 by weight of the total fatty acid content of the emulsion. In some embodiments, the oil contains less than about 0.5% DPAn6 by weight of the total fatty acid content of the emulsion. In some embodiments, the emulsion contains no detectable amount of DPAn6.

[0094] Oil or emulsion containing DHA, or especially containing DHA-EE, may also be substantially free of arachidonic acid (ARA). ARA refers to the compound (all-Z) 5,8,11,14eicosatetraenoic acid (also referred to as (5Z,8Z,11Z,14Z)-icosa-5,8,11,14-tetraenoic acid), as well as any salts or derivatives thereof. Thus, the term ARA encompasses free ARA acid as well as alkyl esters of ARA and triglycerides containing ARA. ARA is a polyunsaturated ω-6 fatty acid. As employed herein, an oil used to obtain the emulsion that is substantially free of ARA refers to an oil in which ARA is less than about 3% by weight of the total fatty acid content of the oil.In some embodiments, the oil comprises less than about 2% ARA by weight of the total fatty acid content of the oil, less than about 1% ARA by weight of the total fatty acid content of the oil, less than about 0.5% ARA by weight of the total fatty acid content of the oil, less than about 0.2% ARA by weight of the total fatty acid content. Petition 870210074972, dated 08 / 16 / 2021, p. 48 / 96 38 / 80 fatty acid content of the oil, or less than about 0.01% ARA by weight of the total fatty acid content of the oil. In some embodiments, the oil has no detectable amount of ARA. As used herein, a substantially ARA-free emulsion refers to an emulsion in which ARA is less than about 3% by weight of the total fatty acid content of the emulsion. In some embodiments, the emulsion includes less than about 2% ARA by weight of the total fatty acid content of the emulsion, less than about 1% ARA by weight of the total fatty acid content of the emulsion, less than about 0.5% ARA by weight of the total fatty acid content of the emulsion, less than about 0.2% ARA by weight of the total fatty acid content of the emulsion, or less than about 0.01% ARA by weight of the total fatty acid content of the emulsion. In some formulations, the emulsion does not contain any detectable amount of ARA.

[0095] PUFA can be added to an emulsion of the present invention as a liquid (e.g., an oil), a solid (e.g., a powder), or a combination thereof.

[0096] PUFAs for use with the present invention may be isolated from any PUFA source comprising at least one PUFA capable of being dispersed in an emulsion. A PUFA for use with the present invention may be, for example, from a microbial source, a plant source, a seed source, an animal source, a fish source, or a combination thereof. Suitable PUFAs and sources of PUFAs for use with the present invention include those described in the US Patent Application Publication. Petition 870210074972, dated 08 / 16 / 2021, p. 49 / 96 39 / 80 number 2009-0023808, which is incorporated herein by reference in its entirety. For example, the PUFAs for use with the present invention may be from an oleaginous microorganism. A PUFA and / or PUFA-containing oil for use with the present invention may also be synthesized.

[0097] In some embodiments, a crude oil containing PUFA (e.g., from a fish, plant, seed and / or microbial source) is refined (to remove phospholipids and free fatty acids), bleached (to remove any colored bodies), enzymatically treated and / or gelled (to remove saturated fats).

[0098] Commercially available PUFAs suitable for use with the present invention include, but are not limited to, Martek DHA™-S Oil (nutritional oil) (Martek Biosciences Corp, Columbia, Md.), Rosemary-Free Martek DHA™-S Oil (rosemary-free nutritional oil) (Martek Biosciences Corporation, Columbia, Maryland), Microalgae DHA™ Oil (microalgae oil) (Martek Biosciences Corporation, Columbia, Maryland), OMEGAPURE® oils (Omega Protein Corp, Houston, Texas), Marinol® Oils (Lipid Nutrition, Wormerveer, NL), Meg-3 oils and powders (Ocean Nutrition Corp, Dartmouth, CA), Evogel (Symrise AG, Holzminden, DE), Marine Oil (Arista Industries, Wilton, Connecticut), and OMEGASOURCE® oils (Food Technology Source, Inc., Raleigh, NC).

[0099] In some embodiments, a PUFA oil is present in an oil-in-water emulsion of the present invention at a concentration of about 5% to about 40%, about 10% to about 30%, about 12% to about 25%, Petition 870210074972, dated 08 / 16 / 2021, p. 50 / 96 40 / 80 approximately 15% to approximately 20%, approximately 12%, approximately 15%, approximately 18%) or 20% by weight of the emulsion.

[00100] In some embodiments, a PUFA oil is present in the discontinuous oil phase of an oil-in-water emulsion of the present invention at a concentration of about 50% to about 99%, about 60% to about 99%, about 70% to about 99%, about 80% to about 99%, about 90% to about 99%, or about 95% to about 99% by weight of the discontinuous oil phase.

[00101] In some embodiments, a water-soluble stabilizer and the PUFA are present in a ratio of about 4:1 to about 1:1 by weight, about 3:1 to about 1:1, about 2:1 to about 1:1, about 3:1, about 2:1, about 3:2, or about 1:1 by weight. Emulsifiers

[00102] As used herein, an emulsifier refers to a material that promotes the stability of an oil-in-water emulsion such that the discontinuous oil phase remains substantially dispersed within the continuous aqueous liquid phase. Generally, an emulsifier is at least partially soluble in either the continuous aqueous liquid phase or the discontinuous oil phase. In some embodiments, an emulsifier is partially soluble in both the continuous aqueous liquid phase and the discontinuous oil phase.

[00103] Suitable emulsifiers for use in the emulsions of the present invention include any emulsifier compatible with the present LC-PUFAs in the emulsions, including natural, modified and synthetic emulsifiers and combinations thereof. Emulsifiers Petition 870210074972, dated 08 / 16 / 2021, page 51 / 96 41 / 80 modified emulsifiers include natural emulsifiers that are modified by chemical, enzymatic and / or physical processes. Emulsifiers particularly suitable for use with the present invention include, but are not limited to, a modified acacia gum (e.g., TICAMULSION® from TIC Gums, White Marsh, Md.), a lecithin, an agar, modified ghatti gum, pectin, carrageenan, xanthan gum, modified starch, in particular, a modified food starch, for example, modified corn starch (available from National & Chemical, Bridgewater, NJ), a modified alginate (for example, alginic acid esters, such as propylene glycol alginate), polyoxyethylene sorbitan, a polyoxyethylene-sorbitan ester (for example, Polysorbate 20, Polysorbate 80, and the like), a sugar ester (for example, sucrose monostearate, and the like), a fatty alcohol (for example, cetostearyl alcohol, cetearyl alcohol, cetylstearyl alcohol and the like), mono- and / or diglycerides, proteins, and combinations thereof. In some embodiments, the emulsifier may be a polymeric hydrocolloid, especially one that originated from a vegetable source.Examples of polymeric hydrocolloids that originate from a vegetable source include vegetable starches, gum arabic (acacia gum), and lignosulfonates, especially food-grade lignosulfonates. In some embodiments, the emulsifier may be a modified acacia gum or a modified starch, such as an acetylated starch or octenyl succinate starch. Examples of commercially available octenyl succinate starch include Cargill EmulTru™ 12674, which is... Petition 870210074972, dated 08 / 16 / 2021, p. 52 / 96 42 / 80 derived from waxy corn starch. In some embodiments, the emulsifier does not have an HLB value. An HLB value refers to the hydrophilic-lipophilic balance value, which is an indication of the degree to which a compound is hydrophilic or lipophilic. In some embodiments, the emulsifying agent has an HLB value of less than 10. In some embodiments, the oil-in-water emulsion does not contain a polyglycerol fatty acid ester. In some embodiments, the emulsifier may be a natural product, such as a natural product extracted from a plant. Therefore, in some embodiments, the emulsifying agent is not a polymeric hydrocolloid, but is preferably a molecule such as, for example, one supplied as quillaja, or Q-Naturale™ (sold by National Starch Food Innovation), which is derived from the quillaja saponaria tree, and combinations of emulsifying agents, such as those listed above.

[00104] In some embodiments, the total concentration of emulsifiers present in an oil-in-water emulsion of the present invention is from 10% to about 25%, about 10% to about 30%, about 12% to about 20%, about 14% to about 18%, about 14%, about 15%, about 16%, about 20%, or about 25% by weight. In some embodiments, the emulsifying agent is present in an amount of less than about 10% by weight of the emulsion, for example, when the emulsifier is lecithin. Excipients

[00105] The thermally stable oil-in-water emulsions of the present invention may comprise one or more excipients. As used herein, an excipient refers to substances useful for... Petition 870210074972, dated 08 / 16 / 2021, p. 53 / 96 43 / 80 combination with a PUFA to provide an oil-in-water emulsion or to provide one or more desired properties to such emulsion. Excipients suitable for use with the present invention meet all the requirements of the current United States and European Pharmacopoeias and various other regulations and standards for pharmaceutical substances, foods, cosmetics and additives. An example of a useful excipient is triacetin (1,2,3-triacetoxypropane; glyceryl triacetate). In general, excipients suitable for use with the present invention are considered safe for human consumption by the US Food and Drug Administration.As used herein, "safe for consumption" refers to excipients, compounds, materials, compositions and / or which, within the scope of sound judgment, are suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other commensurate potential complications with a reasonable risk / benefit ratio. Furthermore, one skilled in the art will recognize that pharmaceutically acceptable excipients may be used in the present invention, including those listed in The Handbook of Pharmaceutical Excipients, 5th ed., The Pharmaceutical Press and American Pharmacists Association, London, United Kingdom and Washington, DC (2006), which is incorporated herein by reference in its entirety.

[00106] In some embodiments, a PUFA contained in an oil-in-water emulsion of the present invention is not substantially detectable by taste and / or smell to a consumer. Thus, an emulsion of the present invention may Petition 870210074972, dated 08 / 16 / 2021, p. 54 / 96 44 / 80 to be ingested by a consumer without any undesirable odor and / or taste. In some embodiments, an emulsion further comprises a flavor masking agent or a flavor masking agent suitable for masking an odor and / or taste of an oil-in-water emulsion of the present invention, or of a product prepared using an oil-in-water emulsion of the present invention. Suitable flavor masking agents for use with the present invention include, but are not limited to, Martek Masker (Martek Biosciences Corp, Columbia, Md.) (supplied by Firmenich (Geneva, Switzerland) and also known as Firmenich Masker), which is a type of vanilla flavoring, Givaudan (Vernier, Switzerland), International Flavors and Fragrances (New York, NY), Sensient Technologies (Milwaukee, WI), and Ogawa Flavors and Fragrances (Tokyo, Japan) and combinations thereof.In some embodiments, the flavor-masking agent is present in an amount of less than 5%, less than 2%, or less than 1% by weight of an emulsion.

[00107] The oil-in-water emulsions of the present invention exhibit an extended shelf life. As used herein, shelf life refers to a period of time within which embodiments of the emulsions can be stored and remain suitable for consumer use. Thus, in addition to particle size stability, the oil-in-water emulsions of the present invention protect and stabilize a PUFA, for example, from oxidation. For example, when administered at doses of 32 mg DHA / 250 mL, the emulsions of the present invention remain stable and organoleptically acceptable for 9 to 12 months of shelf life in dark grape juice such as Concord juice. Petition 870210074972, dated 08 / 16 / 2021, pp. 55 / 96 45 / 80

[00108] In some embodiments, an oil-in-water emulsion of the present invention further comprises an antimicrobial agent, such as potassium sorbate or sodium benzoate or propylene glycol (if it is not already in the emulsion), or mixtures thereof. Such antimicrobial agents may be included in the composition in amounts up to the maximum limit permitted in food and / or beverage compositions. For example, the compositions of the invention may include an antimicrobial agent in an amount between about 0.05 and about 0.1% by weight of the emulsion.

[00109] In some embodiments, an oil-in-water emulsion of the present invention further comprises a preservative. Suitable preservatives for use with the present invention include, but are not limited to, vitamin C, a tocopherol, ascorbic acid or salts thereof (e.g., potassium sorbate), metal chelating agents (i.e., metal chelator) (e.g., ethylenediaminetetraacetic acid (EDTA) and its salts), sodium sulfite and salts thereof (e.g., sodium sulfite, potassium sulfite, and the like), bisulfite, and salts thereof (e.g., sodium bisulfite, etc.), cysteine ​​hydrochloride, polyphosphate (e.g., sodium hexametaphosphate, sodium acid pyrophosphate, sodium mono- and disodium phosphates and the like) and combinations thereof. In some formulations, preservatives are present in amounts of less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%.

[00110] In some embodiments, an oil-in-water emulsion of the present invention further comprises one or more antioxidants. As employed in the present Petition 870210074972, dated 08 / 16 / 2021, p. 56 / 96 46 / 80 document, antioxidants refers to compounds that retard or prevent the oxidation of other chemical species, such as vitamins, pigments, and lipids. Suitable antioxidants for use with the present invention include, but are not limited to, vitamin C (including fat-soluble forms such as ascorbyl palmitate), vitamin E (tocopherols), a polyphenol, a phenol derivative (e.g., butylated hydroxytoluene, butylated hydroxyanisole, t-butylhydroquinone and the like), carnosic acid, lipoic acid, taurine, an aromatic carboxylic acid (e.g., cinnamic acid, benzoic acid, ascorbic acid and the like), salts of an aromatic carboxylic acid (e.g., sodium ascorbate, potassium ascorbate and calcium ascorbate), amino acids that have antioxidant properties, proteins that have antioxidant properties, and combinations thereof.Ascorbate (ascorbic acid) helps to slow down lipid oxidation by reacting with oxygen to remove it from food. Ascorbic acid can also help regenerate oxidized tocopherols to their reduced state, so that tocopherols can continue to function as a free radical scavenger. Thus, the combination of ascorbic acid and tocopherol is especially advantageous, resulting in a synergistic effect in increasing the free radical scavenging capacity of tocopherol.

[00111] Suitable antioxidant polyphenols can be found and extracted from a variety of foods, including plants (e.g., extracts of rosemary, cumin, grape seeds, pine bark, oats, watercress, Petition 870210074972, dated 08 / 16 / 2021, p. 57 / 96 Polyphenols (e.g., basil, ginger, red clover, and similar plants), tea leaves (e.g., green tea, mate (also known as chimarrão or cimarron), and similar plants), fruits (e.g., pomegranate, apple, white cherry, plum, Golji berries, blueberries, tomato, papaya, grapes, and similar plants), vegetables (e.g., alfalfa and similar plants), and cocoa can be synthesized. Exemplary polyphenols include both natural extracts and synthetic compounds.Polyphenols also include, but are not limited to, flavones (e.g., apigenin, luteolin, tangeretin, chrysin, baicalein, scutellarein, wogonin, diosmin, flavoxate, and the like), flavonols (e.g., 3-hydroxyflavone, azaleatin, galangin, gossipectin, kaempferol, isoramethin, morin, myricetin, natsudaidain, paquipodol, quercetin, isoquercitin, quercitrin, rhamnazine, rhamnetin, and the like), flavonols (e.g., (+)-catechin, (-)-epicatechin, (-)-epicatechin gallate, (-)-epigallocatechin gallate), and flavones (e.g., apigenin, luteolin, tangeretin, chrysin, baicalein, scutellarein, wogonin, diosmin, flavoxate, and the like), a flavanone, an isoflavone, a tannin, a stilbene derivative (e.g., resveratrol and the like), an anthocyanin, an anthocyanidin, a proanthocyanidin, gallic acid, curcumin, and combinations thereof.

[00112] An antioxidant may be present in an oil-in-water emulsion of the present invention at a concentration of about 0.01% to about 10%, about 0.02% to about 8%, about 0.05% to about 5%, about 2% to about 20%, about 3% to about 18%, about 4% Petition 870210074972, dated 08 / 16 / 2021, p. 58 / 96 48 / 80 to about 15%, about 5% to about 12%, about 6% to about 10%, or about 7% to about 9% by weight of an emulsion. In some embodiments, an antioxidant is present in an amount of less than about 10%, less than about 5%, or less than about 2% by weight of the emulsion.

[00113] In some embodiments, an oil-in-water emulsion further comprises a flavoring, which may be a synthetic, natural, fruit or botanical flavoring, or a combination thereof. Suitable flavorings for use with the present invention include, but are not limited to, grape, strawberry, raspberry, cherry, almond, citrus, orange, tangerine, lemon, lime, lime-lemon, vanilla, vanilla cream, cocoa, chocolate, coffee, cola, tea, mint, spearmint, wintergreen, menthol, licorice, caramel and combinations thereof.

[00114] In some embodiments, an oil-in-water emulsion further comprises a flavor enhancer which, as used herein, refers to an excipient added to obtain a product with improved flavor or to provide a more pleasant mouthfeel during administration. Non-limiting examples of flavor enhancers suitable for use with the present invention include ribotide and monosodium glutamate.

[00115] In some embodiments, an oil-in-water emulsion additionally comprises a natural or artificial sweetener. Suitable sweeteners include, but are not limited to, sucrose, lactose, fructose, and acesulfame salts (e.g., acesulfame potassium and Petition 870210074972, dated 08 / 16 / 2021, pp. 59 / 96 49 / 80 similar), alitame, aspartame, brazzein, curculin, cyclamic acid and its salts (e.g., sodium cyclamate), dihydrochalcones, glycyrrhizin and its salts, a mogroside, mabinlin, monatin and its salts, monellin, neotame, saccharin and its salts (e.g., sodium saccharin), siamenoside, stevia, stevioside, sucralose, thaumatin, and combinations thereof.

[00116] In some embodiments, a sweetener is present in an oil-in-water emulsion of the present invention at a concentration of about 0.01% to about 20%, about 0.01% to about 1%, about 0.02% to about 15%, about 0.05% to about 10%, about 5% to about 20%, about 0.1% to about 5%, about 0.5% to about 4%, about 1% to about 3%, about 0.01%, about 0.05%, about 0.1%, about 1%, about 5%, or about 10% by weight of the emulsion.

[00117] In some embodiments, an oil-in-water emulsion is sugar-free (i.e., substantially free of a sugar and / or complex carbohydrates) and / or polysaccharides that can be easily converted into a sugar in the oral cavity.

[00118] In some embodiments, an oil-in-water emulsion further comprises a dye. A dye refers to a substance that can be added to an oil-in-water emulsion to improve and / or modify its color or appearance, such as, for example, oligomeric anthocyanins and procyanidins. A dye may also be added to an oil-in-water emulsion as a code or identifier (e.g., to indicate concentration, intended use, and type). Any type of dye (i.e., Petition 870210074972, dated 08 / 16 / 2021, pp. 60 / 96 50 / 80 natural color and / or artificial color, such as FD&C dyes) known as generally considered safe (GRAS) by the FDA, and therefore commonly used in the confectionery trade, or otherwise approved by the FDA for use in pharmaceutical and / or nutraceutical preparations, may be used with the present invention.

[00119] In some embodiments, the discontinuous oil phase further comprises a material selected from: a terpene (e.g., limonene, pinene, and the like), a flavoring oil, a vegetable oil, an essential oil and the like, and combinations thereof. Essential oils suitable for use with the present invention include, but are not limited to, a citrus oil (e.g., lemon, orange, lime, grapefruit, tangerine, bitter orange, and the like), a leaf oil (e.g., mint oil, peppermint oil, and the like), a spice oil (e.g., bergamot oil, rosemary oil, and the like), a seed oil (e.g., linseed oil, cranberry seed oil, and the like), a bark oil, and combinations thereof.

[00120] In some embodiments, an oil-in-water emulsion of the present invention further comprises a thickening agent. Suitable thickening agents for use with the present invention include, but are not limited to, brominated oil (e.g., brominated vegetable oil), ester gum and other wood resins, sucrose diacetate and hexaisobutyrate (SAIB), refined benzoin gum, ganauba wax, benzoate ester of Petition 870210074972, dated 08 / 16 / 2021, pp. 61 / 96 51 / 80 benzyl polyglyceryl, glyceryl tribenzoate and combinations thereof.

[00121] In some embodiments, a thickening agent is present in a continuous aqueous liquid phase at a concentration of about 1% to about 30%, about 2% to about 25%, or about 3% to about 20% by weight of the continuous aqueous liquid phase.

[00122] In some embodiments, an oil-in-water emulsion of the present invention further comprises a water-dispersible or oil-dispersible bioactive. As used herein, water-dispersible bioactive refers to materials that are both dispersible and soluble in water (or an aqueous liquid), and oil-dispersible bioactive refers to materials that are both soluble and dispersible in an oil.Water- and / or oil-dispersible bioactives suitable for use with the present invention include, but are not limited to, an enzyme (e.g., papaya), a carotenoid (e.g., β-carotene, lycopene, astaxanthin, zeaxanthin, lutein, and the like, as well as oxygenated variants thereof), a terpene and / or terpenoid (e.g., eucalyptol, camphor, menthol, citral, and the like), an essential oil (e.g., eugenol, gingerol, avenacoside, and the like), a phenolic acid (e.g., gallic acid, rosmarinic acid, and the like), a flavonoid (e.g., naringin, quercetin, a catechin, an anthocyanin, a coumarin, and the like), a phytoestrogen, a proanthocyanidin, a curcuminoid, a vitamin (e.g., vitamin E, vitamin K, and the like), and combinations thereof. In some formulations, a water-dispersible bioactive is used. Petition 870210074972, dated 08 / 16 / 2021, p. 62 / 96 52 / 80 present in an oil-in-water emulsion, at a concentration of about 0% to about 20%, about 0.5% to about 15%, or about 1% to about 10%, by weight of the emulsion.

[00123] In some embodiments, an oil-in-water emulsion of the present invention further comprises a doubled oil. Doubled oils suitable for use with the present invention include, but are not limited to, 3x, 4x, 5x, 6x, 8x, 10x, 15x, 20x doubled bergamot oil (including bergapten-free bergamot oil), grapefruit oil (including grape oil and high-aldehyde grape juice extracts), lemon, lime, mandarin, orange (as well as orange juice extracts), tangerine, and similar oils, and combinations thereof. Doubled oils suitable for use with the present invention also include washed, distilled, cold-pressed, terpene-free and / or sesquiterpene-free variants of the exemplary doubled oils described above.

[00124] In some embodiments, a folded oil is present in an oil-in-water emulsion of the present invention at a concentration of about 0.1% to about 10%, about 0.2% to about 5%, about 0.3% and about 1%, about 0.5% to about 5%, or about 1% to about 3% by weight of the emulsion.

[00125] In some embodiments, the oil-in-water emulsions of the present invention are substantially free of mono- and / or diglycerides. As used herein, substantially free of mono- and / or diglycerides refers to the oil-in-water emulsions of the present invention comprising about 10% or less, Petition 870210074972, dated 08 / 16 / 2021, pp. 63 / 96 53 / 80 approximately 5% or less, approximately 1% or less, approximately 0.5% or less, approximately 0.1% or less, approximately 0.05% or less, or a detectable concentration of mono- and / or diglycerides, by weight of the emulsion.

[00126] The thermally stable oil-in-water emulsions of the present invention can be used as a component or a functional ingredient, for example, in a food product, a beverage, a plant-based composition, a food supplement, a nutritional product, a pharmaceutical composition (particularly one administered orally or by enteral feeding), and / or a nutraceutical composition. The oil-in-water emulsions of the present invention may be present in such compositions at a concentration suitable to provide a health benefit to a consumer due to the use (e.g., ingestion) of the product.

[00127] Thus, in some embodiments, the method of the present invention comprises daily administration to the needy individual of a thermally stable emulsion of the present invention, or of a product or composition containing the same, comprising a PUFA, particularly DHA, or more especially DHA-EE substantially free of eicosapentaenoic acid (EPA), wherein the DHA is derived from a non-algae source, for example, fish.

[00128] The term individual refers to mammals, such as humans or primates, such as monkeys, apes, orangutans, baboons, gibbons, and chimpanzees. The term individual can also refer to companion animals, for example, dogs and cats, zoo animals; equines, for example, horses, animals that serve as Petition 870210074972, dated 08 / 16 / 2021, pp. 64 / 96 54 / 80 food, for example, cows, pigs, sheep, and disease model animals, for example, rabbits, mice, and rats. The individual can be a human or non-human being. The individual can be of any age. For example, in some embodiments, the individual is a human child, that is, after birth until about 1 year of age; a human infant, that is, a human being between about 1 year of age and 12 years of age; a pubescent human, that is, a human being between about 12 and 18 years of age; or an adult human being, that is, a human being over about 18 years of age. In some embodiments, the individual is an adult, male or female.

[00129] As used in this document, the terms treat and treatment refer to both therapeutic treatment and prophylactic or preventive measures, where the objective is to prevent or slow down (reduce) the undesirable physiological condition or disease, or to obtain the desired clinical results. The term treatment also refers to the relief of symptoms associated with the aforementioned conditions or diseases.

[00130] In some embodiments, the preparation containing the PUFA provided by the emulsion of the present invention is administered continuously. The term continuous or consecutive, as used herein in reference to administration, means that the frequency of administration is at least once a day. It should be noted, however, that the frequency of administration may be greater than once a day and still be continuous or consecutive, for example, twice or even three times a day. Petition 870210074972, dated 08 / 16 / 2021, pp. 65 / 96 55 / 80 four times a day, provided that the dosage levels as specified herein are achieved.

[00131] In some embodiments, an oil-in-water emulsion is a concentrate for appropriate dilution by a local formulator, a filler, distributor, pharmacy, or other entities at the point of distribution and / or use. Oil-in-water concentrates are particularly suitable for products that need to be shipped and / or stored before use.

[00132] As used in this document, a concentrate refers to an oil-in-water emulsion suitable for dilution to produce a final oil-in-water emulsion that has a lower concentration of emulsifier and PUFA. For example, a concentrate may comprise a beverage emulsion concentrate that can be diluted to form a beverage. Specifically, the emulsion concentrate is readily dispersed within a continuous aqueous liquid phase, without further homogenization required. The formation of emulsion concentrates allows for the storage of LC-PUFAs in a stable and compact form for storage as well as transport, before being dosed into a final emulsion form for consumption by a consumer. Furthermore, the homogenization of the emulsion concentrate can be performed on a smaller scale than the homogenization of a final emulsion form to be consumed by a consumer. Thus, lower equipment costs are realized.

[00133] In some embodiments, the concentrate of the present invention can be added to solids or semi-solids. For example, an emulsion concentrate can be Petition 870210074972, dated 08 / 16 / 2021, pp. 66 / 96 56 / 80 added to solid or semi-solid foods or beverages, including but not limited to mayonnaise, whipped toppings, ice cream, yogurt, milkshakes, sauces, fruit concentrates, fruit purees, baby food, specialty coffees such as frappes, etc., and teas, especially iced and specialty coffees and teas containing milk or milk products, such as chai tea, Thai iced tea (cha-yen), and other similar products and combinations thereof.

[00134] In some embodiments, carrier additives, such as potassium sorbate, may be present in the final product at a level that is considered non-functional. For example, a carrier-through level of potassium sorbate of 1-3 ppm in the release of 32 mg of DHA in 256.38 mL of beverage is considered non-functional. In the case of food products, a carrier additive is a substance whose presence in a given product is due only to the fact that the additive was contained in one or more ingredients that went into the making of the product and that it does not serve any technological function in the finished product. Processes for the Preparation of Emulsions

[00135] The oil-in-water emulsions of the present invention can be prepared using any suitable method for mixing an aqueous liquid phase and a discontinuous oil phase to provide an oil-in-water emulsion. In some embodiments, an emulsifier and an aqueous phase material are slowly mixed to provide a stable dispersion, followed by the addition of an optional antioxidant, an optional preservative, an optional pH adjusting agent, and the like. This mixture Petition 870210074972, dated 08 / 16 / 2021, pp. 67 / 96 A 57 / 80 homogeneous aqueous phase is then vigorously combined while slowly adding oil-phase materials (e.g., a PUFA, an optional flavor masking agent, an optional antioxidant, an optional preservative, and others similar) to provide an oil-in-water emulsion. Alternatively, the ingredients of an aqueous liquid, an emulsifier, and an oil phase can be combined simultaneously to form an oil-in-water emulsion.

[00136] The water-soluble stabilizer can be added before, during, and / or after the formation of the oil-in-water emulsion. In some embodiments, a first portion of the water-soluble stabilizer is added to the aqueous phase mixture before emulsification, and a second portion of the water-soluble stabilizer is added to the oil-in-water emulsion after emulsification.

[00137] In some embodiments, a substantially aqueous composition is prepared consisting of water and an emulsifier, and the pH of the aqueous composition is adjusted as described herein by adding an appropriate amount of an acid and / or a base before emulsification.

[00138] In processes involving the sequential addition of ingredients, a substantially homogeneous aqueous phase composition (e.g., a dispersion) is prepared comprising an aqueous liquid, an emulsifier, and one or more optional excipients, and the oil phase ingredients (e.g., a PUFA and one or more optional excipients) are mixed in parallel to provide a substantially homogeneous aqueous phase mixture. Petition 870210074972, dated 08 / 16 / 2021, pp. 68 / 96 58 / 80 oily. The ingredients of the mixed oil phase are then slowly added to the aqueous phase composition while vigorously combining the mixture to provide an oil-in-water emulsion.

[00139] In some embodiments, an emulsifying agent is added to both the aqueous phase and an oil phase before emulsification.

[00140] In some embodiments, an emulsion concentrate comprising a portion of an aqueous liquid, an emulsifier, and a discontinuous oil phase is prepared, and a remaining portion of the aqueous liquid is then added to the emulsion concentrate to form the emulsion.

[00141] In some embodiments, a process comprises: combining water and an emulsifier to provide an aqueous mixture, adding to the aqueous mixture a polyunsaturated fatty acid (preferably an oil comprising a polyunsaturated fatty acid), while mixing so as to obtain an oil-in-water emulsion, and adding to the oil-in-water emulsion a water-soluble stabilizer, selected from a mixture of sodium chloride and a monosaccharide, a mixture of propylene glycol and a monosaccharide, and glycerol, wherein the water-soluble stabilizer is present in a concentration of 20% to 50% by weight of the emulsion to provide a thermally stable oil-in-water emulsion, in which the thermally stable oil-in-water emulsion remains at least partially liquefied at a temperature of -40°C and is free from a change in particle size, free from a change in, or of Petition 870210074972, dated 08 / 16 / 2021, pp. 69 / 96 59 / 80 undesirable organoleptic properties after 9 months of storage at a temperature of -40°C to -15°C.

[00142] In some embodiments, an initially formed oil-in-water emulsion is homogenized by passing the oil-in-water emulsion through a homogenizer one or more times (e.g., once, twice, three times, four or more times) to form a final oil-in-water emulsion. For example, the emulsion may be passed through a homogenizer at a total pressure of 6.8947 kPa / 3.447 kPa second stage, with 5 passes. In another example, the homogenization pressure may be 34.473 kPa total / 5.171 kPa second stage with 2 passes. The pressure and number of passes are determined by the homogenizer scale and type, as well as the desired final particle size.

[00143] In some embodiments, a scraped surface heat exchanger (SSHE) is used with preparations or materials that have a high viscosity, specifically for heating and cooling the preparation and for pasteurization, if desired, for example, with products containing potassium sorbate. In some embodiments, a homogenizer is connected to the scraped surface heat exchanger.

[00144] In some forms, the mixing is carried out under nitrogen cover.

[00145] Having described the invention in general terms, further understanding can be obtained by reference to the examples provided in this document. These examples are provided for illustrative purposes only and are not intended to be limiting. EXAMPLES Petition 870210074972, dated 08 / 16 / 2021, pp. 70 / 96 60 / 80 Example 1

[00146] An oil-in-water emulsion of the present invention was prepared as follows. Potassium sorbate (750 mg) and sodium ascorbate (40.12 g) were added to water (558.54 g) and dissolved. Modified acacia gum (200.59 g) was added to the water mixture, which was then covered and mixed slowly (200 rpm) for 4 to 6 hours. Any foam formed on the surface of the solution was discarded. After slow mixing, the pH of the aqueous mixture was adjusted to pH 4 by adding an appropriate amount of citric acid. The water mixture was used as a base for an oil-in-water emulsion.

[00147] An oil blend was prepared by combining and mixing Martek algae oil (40% DHA oil) (90 g, Martek Biosciences Corp, Columbia, Md.), rosemary extract (157.5 mg; STABILENHANCE® OSR 5% rosemary, 001280, Naturex Inc.), and flavor masking agent (900 mg, Flavor Masking Agent 599469AH, Firmenich, also referred to as Flavor Masking Agent).

[00148] A portion of the water mixture (358.9 g) was placed in a high-shear mixer and combined at 6,100 rpm, while the oil mixture was slowly added to provide an oil-in-water emulsion. Both the water mixture and the oil mixture were at 25°C during mixing. Mixing continued until all of the oil mixture had been emulsified.

[00149] The emulsion was homogenized using a first-stage pressure of 15,000 psi (103.4 MPa) and a second-stage pressure of 2,000 psi (13.79 MPa). The emulsion was Petition 870210074972, dated 08 / 16 / 2021, pp. 71 / 96 The 61 / 80 mixture was repeated through the homogenizer for approximately 30 seconds before collecting a first-pass product. After passing through the homogenizer, the emulsion was cooled by passing the product tube through an ice bath and then collected. The first 10 seconds of product were discarded. The homogenization process was repeated three times to provide an oil-in-water emulsion, which was placed in a high-shear mixer and mixed at 1,500 rpm. Dextrose (27 g) was slowly added to the emulsion, followed by sodium chloride (67.5 g) to provide the final emulsion, which was bottled, purged with nitrogen, and stored at a freezing temperature (-17°C to -21°C). Table 1. Composition of the final emulsion prepared in Example 1 Ingredient Percentage (weight / weight) Water 44% Martek algae oil (40% DHA oil) 15.8% Modified acacia gum 15.8% NaCl 15% Dextrose 6% Sodium ascorbate 3.16% Anhydrous citric acid 0.87% Flavor masking agent 0.16% Potassium sorbate 0.06% Rosemary extracts 0.03% aTICAMULSION® A-2010 Powder (TIC Gums) Petition 870210074972, dated 08 / 16 / 2021, pp. 72 / 96 62 / 80 bFlavor masking agent 599469AH (Firmenich) cRosemary STABILENHANCE® OSR 5% 001280 (Naturex Inc.) Example 2

[00150] An oil-in-water emulsion of the present invention was prepared as follows. Potassium sorbate (730 mg) and sodium ascorbate (38.74 g) were added to water (287.55 g) and dissolved. Dextrose (169.49 g) was added to the water mixture and dissolved, being heated if necessary. The water mixture was then cooled (if necessary) and propylene glycol (193.7 g) was added, and the resulting water mixture was thoroughly combined. Modified gum arabic (159.8 g) was then added to the water mixture, which was then covered and mixed slowly (200 rpm) for 4 to 6 hours. Any foam formed on the surface of the solution was discarded. After slow mixing, the pH of the aqueous mixture was adjusted to pH 4 by adding an appropriate amount of citric acid.

[00151] A portion of the water mixture (394.9 g) was placed in a high-shear mixer and combined at 6,100 rpm, while the oil mixture was slowly added to provide an oil-in-water emulsion. Both the water mixture and the oil mixture were at 25°C during mixing.

[00152] The mixing continued until the entire oil mixture was emulsified. The emulsion was homogenized using a front pressure of 15,000 psi (103.4 MPa) and a back pressure of 2,000 psi (13.79 MPa). A Petition 870210074972, dated 08 / 16 / 2021, pp. 73 / 96 The 63 / 80 emulsion was cycled through the homogenizer for approximately 30 seconds before collecting a first pass of product. After passing through the homogenizer, the emulsion was cooled by passing the product hose through an ice bath and then collected. The first 10 seconds of product were discarded. The homogenization process was repeated three times to provide the oil-in-water emulsion, which was packaged, purged with nitrogen, and maintained at a freezing temperature (-17°C to -21°C). Table 2. Composition of the final emulsion prepared in Example 2. Ingredient Percentage (weight / weight) Water 29.7% Propylene Glycol 20% Dextrose 17.5% Modified Acacia Gum 16.5% Martek Algae Oil (40% DHA oil) 12% Sodium Ascorbate 4% Anhydrous Citric Acid 0.5% Flavor Masking Agent 0.2% Rosemary Extracts 0.04% Potassium Sorbate 0.08% aTICAMULSION® A-2010 Powder (TIC Gums) bFlavor masking agent 599469AH (Firmenich) cRosemary STABILENHANCE® OSR 5% 001280 (Naturex) (Inc.) Example 3 Petition 870210074972, dated 08 / 16 / 2021, pp. 74 / 96 64 / 80

[00153] An oil-in-water emulsion of the present invention was prepared as follows. Potassium sorbate (730 mg) and sodium ascorbate (38.74 g) were added to water (287.55 g) and dissolved. Modified gum arabic (159.8 g) was then added to the water mixture, which was then covered and combined slowly (200 rpm) for 4 to 6 hours. Any foam formed on the surface of the solution was discarded. After slow combination, the pH of the aqueous mixture was adjusted to pH 4 by adding an appropriate amount of citric acid. Glycerol (162 g) was then added to the water mixture and the water mixture was thoroughly combined.

[00154] A portion of the water mixture (216.3 g) was placed in a high-shear mixer and combined at 6,100 rpm, while the oil mixture was slowly added to provide an oil-in-water emulsion. Both the water mixture and the oil mixture were at 25°C during the combination. Mixing continued until the entire oil mixture was emulsified.

[00155] The emulsion was homogenized using a front pressure of 15,000 psi (103.4 MPa) and a back pressure of 2,000 psi (13.79 MPa). The emulsion was cycled through the homogenizer for approximately 30 seconds before collecting a first-pass product. After passing through the homogenizer, the emulsion was cooled by passing the product hose through an ice bath and then collected. The first 10 seconds of product were discarded. The homogenization process was repeated three times to Petition 870210074972, dated 08 / 16 / 2021, pp. 75 / 96 65 / 80 provides an oil-in-water emulsion that has been packaged, purged with nitrogen, and kept at a freezing temperature (-17°C to -21°C). 3. Composition of the final emulsion prepared in Example 3. Ingredient Percentage (weight / weight) Glycerin 36% Water 28% Martek Algae Oil (40% DHA oil) 15% Modified Acacia Gum 14% Sodium Ascorbate 4% Anhydrous Citric Acid 0.78% Double Oil 0.33% Flavor Masking Agent 0.2% Rosemary Extract 0.2% Mixed Tocopherols 0.2% Potassium Sorbate 0.08% aTICAMULSION® A-2010 Powder (TIC Gums) bFlavor masking agent 599469AH (Firmenich) cRosemary STABILENHANCE® OSR 5% 001280 (Naturex) Inc.) dTOCOBLEND® L70 (Vitablend BV) Example 4

[00156] The following emulsions were prepared in a similar manner to the emulsion in Example 3. Table 4. Composition of a final emulsion. Ingredient Percentage Petition 870210074972, dated 08 / 16 / 2021, pp. 76 / 96 66 / 80 (weight / weight) Water 33% Glycerin 31.03% Modified food starcha 15% Martek algae oil (40% DHA oil) 10% Citric acid, anhydrous 5.4% Sodium ascorbate 2.5% Trisodium citrate, anhydrous 2.5% Sodium hexametaphosphate 0.15% Flavor masking agentb 0.15% Rosemary extractc 0.1% Mixed tocopherolsd 0.1% Potassium sorbate 0.07% aCargill EmulTru™ 12674 bFlavor Masking Agent 599469AH (Firmenich) cRosemary STABILENHANCE® OSR 5% 001280 (Naturex Inc. ) dTOCOBLEND® L70 (Vitablend BV) Table 5. Composition of a final emulsion. Ingredient Percentage (weight / weight) Water 34.23% Glycerin 30.43% Modified food starch 0.15% Martek algae oil (40% DHA oil) 10% Citric acid, anhydrous 4.8% Sodium ascorbate 2.5% Trisodium citrate, anhydrous 2.5% Petition 870210074972, dated 08 / 16 / 2021, pp. 77 / 96 67 / 80 Sodium hexametaphosphate 0.15% Flavor masking agent 0.15% Mixed tocopherols 0.1% Rosemary extracts 0.1% Potassium sorbate 0.07% aNational Starch & Chemical (Purity Gum 2000) bFlavor Masking Agent 599469AH (Firmenich) cTOCOBLEND® L70 (Vitablend BV) dRosemary STABILENHANCE® OSR 5% 001280 (Naturex (Inc.) Table 6. Composition of a final emulsion. Ingredient Percentage (weight / weight) Water 30% Glycerin 29.03% Modified food starcha 15% Martek algae oil (40% DHA oil) 15% Citric acid, anhydrous 5.4% Sodium ascorbate 2.5% Trisodium citrate, anhydrous 2.5% Sodium hexametaphosphate 0.15% Flavor masking agentb 0.15% Mixed tocopherolssc 0.1% Rosemary extractsd 0.1% Potassium sorbate 0.07% aTICAMULSION® A2010 Powder (TIC Gums) bFlavor Masking Agent 599469AH (Firmenich) Petition 870210074972, dated 08 / 16 / 2021, pp. 78 / 96 68 / 80 cTOCOBLEND® L70 (Vitablend BV) dRosemary STABILENHANCE® OSR 5% 001280 (Naturex Inc.) Example 5

[00157] The stability of the oil-in-water emulsions prepared in Examples 1 and 3 was tested by maintaining the emulsions at a temperature of -17°C for a period of 5, 6, 8, or 10 months, followed by thawing the oil-in-water emulsions at a temperature of 4°C and quantitatively evaluating the properties of the emulsions.

[00158] Oil-in-water emulsions (10 mL) were placed in a 15 mL container of an ice cream maker (fast freezing) or in a normal freezer (slow freezing) and stored at -17°C for up to 10 months. The particle size of the oil phase was measured after the designated period at -17°C and is reported in Tables 7 and 8 below. Particle size was measured using a Malvern Mastersizer Hidro 2000S (Malvern Instruments, Ltd., Worcestershire, UK). Individual samples of the oil-in-water emulsions were removed from the freezer at the designated interval, thawed, and the particle size and organoleptic characteristics (using juice model tests) of the emulsions were determined. Table 7. Particle size data as a function of emulsion stability for the oil-in-water emulsions prepared in Example 1. Sample name d(0.5) D[4.3] Uniformity* Initial at 4°C 0.127 0.136 0.338 Rapid Freezing, 5 months 0.129 0.137 0.305 Petition 870210074972, dated 08 / 16 / 2021, pp. 79 / 96 69 / 80 Slow Freezing, 5 months 0.126 0.134 0.318 Rapid Freezing, 6 months 0.127 0.136 0.34 Slow Freezing, 6 months 0.127 0.136 0.34 Rapid Freezing, 8 months 0.127 0.137 0.338 Slow Freezing, 8 months 0.126 0.135 0.335 Rapid Freezing, 10 months 0.122 0.132 0.373 Slow Freezing, 10 months 0.127 0.136 0.336 * A lower uniformity number corresponds to a higher quality emulsion. Table 8. Particle size data as a function of emulsion stability for the oil-in-water emulsions prepared in Example 3. Sample Name d(0.5) D[4.3] Uniformity* Initial at 4°C 0.154 0.17 0.413 Rapid Freezing, 5 months 0.151 0.168 0.418 Slow Freezing, 5 months 0.148 0.169 0.472 Rapid Freezing, 6 months 0.175 0.193 0.423 Slow Freezing, 6 months 0.175 0.194 0.424 Rapid Freezing, 8 months 0.168 0.186 0.421 Slow Freezing, 8 months 0.164 0.182 0.422 Petition 870210074972, dated 08 / 16 / 2021, pages 80 / 96 70 / 80 Fast Freezing, 10 months 0.164 0.182 0.42 Slow Freezing, 10 months 0.169 0.187 0.422

[00159] As shown in Tables 7 and 8, the particle size of the oil-in-water emulsions did not change significantly even after 10 months of storage at -17°C.

[00160] The organoleptic qualities of the oil-in-water emulsions were determined by a taste test with an expert jury. Briefly, the emulsions (32 mg DHA) were mixed with 8 ounces (236.6 mL) of green grape or Concord grape juice, microwaved, and tested using the expert jury. The formulations of Examples 1 and 3 had good organoleptic quality. Example 6

[00161] The freeze-thaw stability of the oil-in-water emulsions prepared in Example 4, Table 5, was tested by maintaining the emulsions at a temperature of -17°C for a period of 5 weeks. Each week the emulsion was thawed at a temperature of 4°C and tested. The particle size of the oil phase was measured using a Malvern Mastersizer Hidro 2000S (Malvern Instruments, Ltd., Worcestershire, United Kingdom) after the designated number of freeze-thaw cycles and is presented in Table 9 below. The data are graphically represented in Figure 1. Table 9. Particle size data as a function of emulsion stability for the oil-in-water emulsions prepared in Example 1. Petition 870210074972, dated 08 / 16 / 2021, pp. 81 / 96 71 / 80 Number of freeze-thaw cycles d(0.5) (pm) D[3.2] (pm) D[4.3] (pm) % of Results 0.01 pm - 0.36 pm Uniformity* 0 0.122 0.115 0.129 100 0.289 1 0.123 0.114 0.131 100 0.322 2 0.121 0.112 0.130 100 0.330 3 0.120 0.111 0.130 100 0.342 4 0.121 0.11 0.132 100 0.364 5 0.121 0.11 0.132 100 0.364 * A lower uniformity number corresponds to a higher quality emulsion.

[00162] As shown in Table 9, the oil-in-water emulsion of Example 4, Table 5, went through 5 freeze-thaw cycles without a change in particle size. Example 7

[00163] The freeze-thaw stability of the oil-in-water emulsions prepared in Example 4, Tables 4 and 6, was tested by maintaining the emulsions at a temperature of -17°C for a period of 5 weeks. For the stability test, the frozen oil-in-water emulsions were thawed overnight once a week at a temperature of 4°C, tested, and frozen again.

[00164] Oil-in-water emulsions were tested using a drinking ring test. Briefly, 540 g of water, 0.3 g of potassium sorbate, and 60 g of sucrose were added to a container and mixed for 2 minutes. Citric acid was added to adjust the pH of the Petition 870210074972, dated 08 / 16 / 2021, pp. 82-96 72 / 80 solution to 4.0. An equivalent amount of the emulsion to be tested was added to the beaker during mixing. The mixture was then poured into two round Boston glass flasks. One was placed horizontally and the other vertically. The flasks were monitored for 10 days at room temperature. Rings were identified by slightly tilting the solution in the glass flasks and examining for the presence of a visible ring at the top of the solution inside the container. If no ring was formed, the emulsion passed the ring test. The measurement of the oil phase particle size and the ring test were performed after the designated number of freeze-thaw cycles and are presented in Tables 10 and 11 below. Table 10. Particle size data as a function of freeze-thaw emulsion stability for the oil-in-water emulsions prepared in Example 4, Table 4. Weeks d(0.5) (pm) D[3.2] (pm) D[4.3] (pm) % of Results 0.01 pm 0.36 pm Uniformity* Ring Test (pass / fail) 0 0.120 0.113 0.128 100 0.306 Passed 1 0.125 0.115 0.134 100 0.338 Passed 2 0.119 0.107 0.130 100 0.373 Passed 3 0.119 0.107 0.130 100 0.374 Passed 4 0.119 0.107 0.130 100 0.373 Passed 5 0.121 0.107 0.130 100 0.372 Passed Table 11. Particle size data as a function of emulsion stability in freeze-thaw cycle. Petition 870210074972, dated 08 / 16 / 2021, pp. 83 / 96 73 / 80 thawing for the oil-in-water emulsions prepared in Example 4, Table 6. Weeks d(0.5) (pm) D[3.2] (pm) D[4.3] (pm) % of Results 0.01 pm 0.36 pm Uniformity of* Ring Test (pass / fail) 0 0.138 0.126 0.146 100 0.326 Passed 2 0.141 0.129 0.148 100 0.300 Passed 5 0.133 0.121 0.141 100 0.329 Passed

[00165] As shown in Tables 10 and 11, the formulations of Example 4, Tables 4 and 6, exhibit consistent particle size and passed the ring test after several freeze-thaw cycles. Example 8 Sensory Score of a Six-Month Emulsion

[00166] The descriptive analysis (DA) test was performed to obtain a sensory score over six months for the emulsions obtained according to the invention as follows. For each preparation, the strength of each attribute was rated on an intensity scale of 0-15 points, with 0 = none and 15 = very strong. Martek DA judges who were trained and experienced in aroma and flavor analysis evaluated Concord grape juice. The compositions evaluated were as follows.

[00167] The emulsions were prepared from starch obtained from two different starch suppliers (Cargill CS starch (EmulTru™ 12674) and National Starch & Chemical NS starch), and a supplier of modified acacia gum (TICAMULSION® A-2010 Powder, (TIC Gum MGA)) and Petition 870210074972, dated 08 / 16 / 2021, pp. 84 / 96 74 / 80 at a cooling temperature of -20°C. A total of three emulsions were prepared as follows: - CS-20: Cargill starch used as an emulsifier; - MGA-20: TIC Gum, modified acacia gum used as an emulsifier; and - NS-20C: Modified food starch from National Starch & Chemical used as an emulsifier.

[00168] A rating of 2 is the highest score to pass the test; none of the emulsions received a sensory score higher than 2 during the six-month evaluation period.

[00169] The CS-20, MGA-20, and NS-20C emulsions prepared as shown in Table 4 above were dosed into Welch Concord grape juice at 32 mg DHA / 250 g of juice. The dosed grape juice was processed with a microThermics at a preheating temperature of 250°F (121.1°C) and a filling temperature of 185°F (85°C). The grape juice was processed in PET containers, cooled, and stored at room temperature. The following samples were tested: (1) Control 1 containing grape juice dosed with the emulsion prepared by the same formulation as above, but in which a high oleic sunflower oil (HOSO) replaced the DHA™-S oil; (2) Control 2 containing grape juice processed by microThermics without the addition of any emulsion; (3) Sample 1 containing grape juice dosed with CS-20C emulsion and processed by microThermics; Petition 870210074972, dated 08 / 16 / 2021, pages 85 / 96 75 / 80 (4) Sample 2, containing grape juice dosed with NS-20C emulsion and processed by MicroThermics; (5) Sample 3 containing juice dosed with NS-20C emulsion and processed by microThermics.

[00170] The emulsions were evaluated for the presence of suspected aromas in the juice under two conditions: (a) sample storage at 32.2°C for 16 weeks (which accelerated the development of any aroma) and (b) sample storage at room temperature for 3 months. As above, the sensory classification of the DA sensory spectrum is a fifteen-point intensity scale on the spectrum where 0 = none and 15 = strong intensity. Neither sample accelerated (stored at) 32.2°C and at the end of 16 weeks, nor did the samples stored at room temperature and at the end of 3 months have an intensity scale classification greater than 2. Example 9 Differential Test (DFC)

[00171] A Different Control Test (DFC) was conducted as follows. The DFC test was conducted with Martek employees. The jurors were instructed to compare the unenriched sample (control) with the other variables fortified with DHA in order to determine if there is a difference between them. They were also instructed to measure the size of the difference, if any, on a 7-point scale of 0-6, with 0 being no difference found and 6 being a very large difference found. There was little or no change in the DFC test results for either sample after three months at room temperature. Petition 870210074972, dated 08 / 16 / 2021, pp. 86 / 96 76 / 80 Example 10 Thermally stable emulsion based on glycerin Q-Naturale™1

[00172] To obtain a thermally stable emulsion based on Q-Naturale™ glycerin, the following procedure was used. Q-Naturale™ from National Starch Food Innovation was employed. Glycerin was added to the Q-Naturale™ solution using a high-shear mixer at 6,000 rpm. The dry ingredients (sodium hexametaphosphate, sodium ascorbate, anhydrous trisodium citrate, potassium sorbate, and citric acid) were mixed at 6,000 rpm in the high-speed mixer. Martek DHA™-S oil and other oil components (flavor masking agent, rosemary extract, and tocoblend) were mixed at 6,000 rpm in the high-shear mixer. The preparation was mixed until all the oil was emulsified and no longer clinging to the edges (1 minute). The emulsion was then homogenized using 10,000 psi (68.95 MPa) for the first stage and 1,000 psi (6.895 MPa) for the second stage.The product was cooled by applying a cold water bath to the product hose and a circulating water bath in the homogenizer. The first 10 seconds of the products produced were discarded. A 30-second cycling time was allowed before collecting the next products that passed through. The products were homogenized during 8 passes. The resulting emulsion was packaged, purged with nitrogen, and placed in the freezer. The composition of the final emulsion is shown in Table 12. Table 12. Glycerin Emulsion - Q-Naturale™ Petition 870210074972, dated 08 / 16 / 2021, pages 87 / 96 77 / 80 Ingredient Percentage by weight Grams Q Naturale™ 200, 20-22 Brix 30.00 150.00 Glycerin 29.03 145.15 Sodium hexametaphosphate 0.15 0.75 Sodium ascorbate 2.50 12.50 Anhydrous trisodium citrate 2.50 12.50 Potassium sorbate 0.07 0.35 Anhydrous citric acid 5.40 27.00 Martek DHA™-S Rosemary Sun 30.00 150.00 Flavor masking agent 599469AH (Firmiech) 0.15 0.75 Rosemary STABILENHANCE® OSR 5% 001280 0.10 0.50 Tocoblend™70 IP 0.10 0.50 Total 100.00 500.00

[00173] Martek DHA™-S Rosemary Sun is a rosemary extract containing sunflower lecithin. Q-Naturale™ is a natural oil emulsifier derived from the quillaja saponaria tree and is commercially available from National Starch Food Innovation. It is a molecule that performs similarly to gum arabic (acacia gum) in sensory evaluations and creates emulsions with opacity levels similar to gum arabic and starch. However, it is stable over a wide pH and temperature range.

[00174] The particle size of the freezable emulation produced above was tested for uniformity and particle size initially, after preparation, and after three months of storage. The results are shown in Table 13. Petition 870210074972, dated 08 / 16 / 2021, pp. 88 / 96 78 / 80 Table 13 Average Uniformity (μm) % 0.36-5 μm % 0.01-0.36 μm 0 months 3 months 0 months 3 months 0 months 3 months 0 months 3 months 0.262 0.262 0.112 0.112 0 0 100 100

[00175] The emulsion scored a passing grade when subjected to the Ring Test. A Heated Fill Test was conducted with a preheat temperature of 150°F (65.6°C) / final heating of 200°F (93.3°C) / filling temperature of 175°F (79.4°C). The emulsion was measured in Welch's grape juice. The dosage level of the emulsion was 32 mg DNA / 250 g. No creaming was observed as a result of the Heated Fill Test. Example 11

[00176] Formulation of a modified starch based on a thermally stable emulsion with propylene glycol

[00177] A thermally stable emulsion-modified starch based on propylene glycol was prepared as shown in Table 14. Table 14: Modified starch and propylene glycol Ingredient Percentage by weight Grams Cargill EmulTru™ 12674 Starch 15.00 75 Propylene glycol 31.03 155.15 Sodium hexametaphosphate 0.15 0.75 Sodium ascorbate 2.50 12.50 Anhydrous trisodium citrate 2.50 12.50 Potassium sorbate 0.07 0.35 Anhydrous citric acid 5.40 27.00 DHA-S Rosemary Sun 10.00 50.00 Petition 870210074972, dated 08 / 16 / 2021, pages 89 / 96 79 / 80 Flavor masking agent 599469AH 0.15 0.75 Rosemary STABILENHANCE® OSR 5% 001280 0.10 0.50 Tocoblend™ 70 IP 0.10 0.50 Water (deionized) 33.00 165 Total 100.00 500.00 Example 12

[00178] Formulation of a modified starch based on a thermally stable emulsion with propylene glycol, glycerin and triacetin

[00179] A modified starch based on a thermally stable emulsion with propylene glycol, triacetin and glycerin was prepared as shown in Table 15. Table 15

[00180] Modified starch, propylene glycol, triacetin and glycerin Ingredient Percentage by weight Grams Cargill EmulTru™ 12674 Starch 15.00 75 Glycerin 4.00 20 Propylene glycol 14.03 70.15 Triacetin 13.00 65 Sodium hexametaphosphate 0.15 0.75 Sodium ascorbate 2.50 12.50 Anhydrous trisodium citrate 2.50 12.50 Potassium sorbate 0.07 0.35 Anhydrous citric acid 5.40 27.00 Petition 870210074972, dated 08 / 16 / 2021, pages 90 / 96 80 / 80 DHA-S Rosemary Sun 10.00 50.00 Flavor masking agent 599469AH 0.15 0.75 Rosemary STABILENHANCE® OSR 5% 001280 0.10 0.50 Tocoblend™ 70 IP 0.10 0.50 Water (deionized) 33.00 165 Total 100.00 500.00 CONCLUSION

[00181] All the various embodiments or options described in this document can be combined in any and all variations. Although the invention has been particularly illustrated and described with reference to some embodiments thereof, it will be understood by those skilled in the art that these have been presented only by way of example, and not as a limitation, and various alterations in form and details can be made without departing from the spirit and scope of the invention. Thus, the breadth and scope of the present invention should not be limited by any of the exemplary embodiments described above, but should be defined only in accordance with the following claims and their equivalents.

[00182] All documents cited herein, including journal articles or abstracts, published or corresponding U.S. or foreign patent applications, foreign patents or issued patents, or any other documents, are incorporated herein in their entirety by reference, including all data, tables, figures and text presented in the cited documents. Petition 870210074972, dated 08 / 16 / 2021, pages 91 / 96

Claims

1 / 2 CLAIMS 1. Thermally stable oil-in-water emulsion characterized by comprising: an oil comprising a polyunsaturated fatty acid present in a concentration of 5% to 40% by weight of the emulsion, water present in a concentration of 20% to 60% by weight of the emulsion, an emulsifier present in a concentration of 10% to 30% by weight of the emulsion, a water-soluble stabilizer selected from glycerol in a concentration of 25% to 46% by weight of the emulsion, and wherein the thermally stable oil-in-water emulsion is flowable at a temperature of -40°C, and is free from a variation in particle size after 9 months of storage at a temperature of -40°C to -15°C, and wherein the emulsifier is at least one selected from the group consisting of a modified acacia gum and a modified food starch.

2. Emulsion, according to claim 1, characterized in that the emulsion is flowable at a temperature of -80°C.

3. Emulsion, according to claim 1, characterized in that the thermally stable oil-in-water emulsion is free from a change in particle size after 10 freeze-thaw cycles.

4. Emulsion, according to claim 1, characterized by further comprising polypropylene glycol in Petition 870210100103, dated 10 / 29 / 2021, page 14 / 16 2 / 2 at a concentration of 10% to 30% by weight of the emulsion.

5. Emulsion, according to claim 1, characterized in that the polyunsaturated fatty acid is at least one selected from the group consisting of α-linolenic acid, γ-linolenic acid, linoleic acid, conjugated linoleic acid, arachidonic acid, ω-3 docosapentaenoic acid, ω-6 docosapentaenoic acid, eicosapentaenoic acid, docosahexaenoic acid and oxylipins. Petition 870210100103, dated 10 / 29 / 2021, p. 15 / 16

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