US20130316061A1 - Oxidatively stable polyunsaturated fatty acid containing oil - Google Patents

Oxidatively stable polyunsaturated fatty acid containing oil Download PDF

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Publication number
US20130316061A1
US20130316061A1 US13/694,134 US201213694134A US2013316061A1 US 20130316061 A1 US20130316061 A1 US 20130316061A1 US 201213694134 A US201213694134 A US 201213694134A US 2013316061 A1 US2013316061 A1 US 2013316061A1
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oil
ppm
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oil according
tocopherol
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Michael Lenn Stefanski
Xuejun Tang
Kristine Sheila Crawford
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DSM IP Assets BV
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DSM IP Assets BV
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Priority to US13/694,134 priority Critical patent/US20130316061A1/en
Assigned to DSM IP ASSETS B.V. reassignment DSM IP ASSETS B.V. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: CRAWFORD, Kristine Sheila, TANG, XUEJUN, STEFANSKI, MICHAEL LENN
Publication of US20130316061A1 publication Critical patent/US20130316061A1/en
Priority to US14/446,218 priority patent/US20150017304A1/en
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/007—Other edible oils or fats, e.g. shortenings, cooking oils characterised by ingredients other than fatty acid triglycerides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23D—EDIBLE OILS OR FATS, e.g. MARGARINES, SHORTENINGS, COOKING OILS
    • A23D9/00—Other edible oils or fats, e.g. shortenings, cooking oils
    • A23D9/06—Preservation of finished products
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
    • A23L27/82—Acid flavourants
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
    • A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
    • A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
    • A23L3/3463—Organic compounds; Microorganisms; Enzymes
    • A23L3/3544—Organic compounds containing hetero rings
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/115—Fatty acids or derivatives thereof; Fats or oils
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0007—Organic substances
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0085—Substances of natural origin of unknown constitution, f.i. plant extracts
    • C—CHEMISTRY; METALLURGY
    • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
    • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
    • C11B5/00—Preserving by using additives, e.g. anti-oxidants
    • C11B5/0092—Mixtures
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/02—Antioxidant

Definitions

  • the invention relates to an oil comprising one or more polyunsaturated fatty acids having at least 4 double bonds.
  • Oils containing polyunsaturated fatty acids may oxidize and become rancid during storage. This results in unpleasant flavors and odors in the oil as well as in the products in which such oils are incorporated.
  • a PUFA with 4 or more C—C double bonds may be less oxidatively stable than a PUFA with fewer double bonds.
  • metals such as, iron, may increase the rate at which a PUFA containing oil oxidizes.
  • FIG. 1 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 7b, 7c, 8e, and 8f.
  • FIG. 2 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 7d, 7e, and 8b.
  • FIG. 3 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 7f, 7g, and 8c.
  • FIG. 4 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 7h, 7i, and 7j.
  • FIG. 5 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 7c, 7k, and 8a.
  • FIG. 6 is a graph depicting the concentration of iron (ppb) contained in oils substantially free of lecithin (examples 7a, 7c, 7h, 8a-c, and 80 versus an oil containing mixed tocopherol, ascorbyl palmitate, and lecithin (example 7b).
  • FIG. 7 is a graph depicting the fishy aroma sensory value over time of oils according to examples 7a, 8d, and 7m.
  • an oil comprising (i) at least 30 wt. % of one ore more polyunsaturated fatty acids having at least 4 double bonds; (ii) at least one first antioxidant; and (iii) less than about 1000 ppm lecithin.
  • an oil comprising (i) at least 30 wt. % of one or more polyunsaturated fatty acids having at least 4 double bonds; (ii) at least one first antioxidant; and (iii) less than about 750 ppm of a second antioxidant chosen from an ascorbic acid derivative.
  • an oil comprising (i) at least one polyunsaturated fatty acid having at least 4 double bonds; (ii) at least one first antioxidant; and (iii) less than about 30 ppb iron.
  • the oil comprises at least 30 wt. % of one or more polyunsaturated fatty acids having at least 4 double bonds.
  • an oil comprising (i) at least about 30 wt. % of one or more polyunsaturated fatty acids having at least 4 carbon-carbon double bonds; (ii) at least one first antioxidant; and (iii) means for improving oxidative stability, wherein said oil has a fishy aroma of less than 1.5.
  • wt. % of one or more polyunsaturated fatty acids having at least 4 double bonds is expressed with respect to the sum weight of the total fatty acids in the oil.
  • wt. % of one or more polyunsaturated fatty acids refers to the weight percentage of the sum of all polyunsaturated fatty acids having at least 4 double bonds present in the oil.
  • wt. % of a specific fatty acid or any combination of specific fatty acids is expressed with respect to the sum weight of the total fatty acids in the oil.
  • lecithin means less than or equal to about 150 ppm, about 15 ppm, about 10 ppm, about 5 ppm, about 1 ppm, and about 0 ppm.
  • essentially free when used to describe an ascorbic acid derivative means less than or equal to about 3 ppm, to about 2.5 ppm, to about 2 ppm, to about 1.5 ppm, to about 1 ppm, to about 0.5 ppm, to about 0.
  • lecithin when used herein includes, for example, sunflower lecithin, soy lecithin, egg lecithin, and mixtures thereof.
  • the oil described herein comprises less than about 1200 ppm lecithin, less than about 1000 ppm lecithin, less than 750 ppm lecithin, less than about 500 ppm lecithin, less than about 250 ppm lecithin, or less than about 200 ppm lecithin.
  • the oil described herein is free of lecithin. In another embodiment, the oil described herein is essentially free of lecithin.
  • the oil described herein has an iron content of less than about 30 ppb or more particularly less than or equal to about 25 ppb, 20 ppb, 15 ppb, 10 ppb, 5 ppb, 1 ppb, 0.5 ppb, or 0.2 ppb. In another embodiment, the iron content is 0 ppb.
  • antioxidant when used herein includes, for example, ascorbic acid derivatives, tocopherol, green tea extract, and/or mixtures thereof.
  • the antioxidant is a first antioxidant chosen from tocopherol, green tea extract, and/or mixtures thereof. In another embodiment, the antioxidant is a second antioxidant chosen from ascorbic acid derivatives.
  • ascorbic acid derivative when used herein includes, for example, ascorbic acid, ascorbyl palmitate, ascorbyl stearate, and/or mixtures thereof. In one embodiment, the ascorbic acid derivative is ascorbyl pamitate.
  • the ascorbic acid derivative is present in the oil described herein in an amount ranging from about 0 ppm to about 50 ppm, from about 0 ppm to about 250 ppm, from about 0 ppm to about 300 ppm, from about 0 ppm to about 400 ppm, from about 0 ppm to about 750 ppm, from about 50 ppm to about 250 ppm, from about 50 ppm to about 300 ppm, from about 50 ppm to about 400 ppm, from about 50 ppm to about 750 ppm, from about 250 ppm to about 750 ppm, from about 250 ppm to 400 ppm, from about 250 ppm to about 300 ppm, from about 300 ppm to about 400 ppm.
  • the oil described herein comprises less than about 750 ppm ascorbic acid derivative, less than about 400 ppm ascorbic acid derivative, less than about 300 ppm ascorbic acid derivative, less than about 250 ppm ascorbic acid derivative, or less than about 50 ppm ascorbic acid derivative.
  • the oil described herein is essentially free of an ascorbic acid derivative.
  • the oil described herein is essentially free of ascorbyl palmitate.
  • the oil described herein comprises about 250 ppm ascorbic acid derivative.
  • the oil described herein comprises about 250 ppm ascorbyl palmitate.
  • the oil described herein is free of ascorbic acid derivative.
  • the oil is free of ascorbyl palmitate.
  • the oil described herein is essentially free of an ascorbic acid derivative and essentially free of lecithin. In a further embodiment, the oil described herein is essentially free of ascorbyl palmitate and essentially free of lecithin. In another embodiment, the oil is free of an ascorbic acid derivative and free of lecithin. In yet another embodiment, the oil is free of ascorbyl palmitate and free of lecithin.
  • the at least one first antioxidant is a tocopherol. In another embodiment, the at least one first antioxidant is an addition tocopherol. In a still a further embodiment, the at least one first antioxidant is a mixed tocopherol. In another embodiment, the at least one first antioxidant is dl- ⁇ -tocopherol, d- ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol and ⁇ -tocotrienol, or a mixture thereof.
  • tocopherol any isomer of tocopherol (or mixture thereof), including but not limited to dl- ⁇ -tocopherol (i.e., synthetic tocopherol), d- ⁇ -tocopherol (i.e., natural tocopherol), ⁇ -, ⁇ -, and ⁇ -tocopherol dl- ⁇ -tocopherol, and ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol and ⁇ -tocotrienol.
  • dl- ⁇ -tocopherol i.e., synthetic tocopherol
  • d- ⁇ -tocopherol i.e., natural tocopherol
  • ⁇ -, ⁇ -, and ⁇ -tocopherol dl- ⁇ -tocopherol dl- ⁇ -tocopherol
  • ⁇ -tocotrienol ⁇ -tocotrienol
  • ⁇ -tocotrienol ⁇ -tocotrienol
  • the oil described herein may comprise tocopherol in an amount ranging from about 900 ppm to about 3400 ppm, from about 900 ppm to about 2400 ppm, from about 900 ppm to about 2000 ppm, from about 900 ppm to about 1700 ppm, from about 900 ppm to about 1400 ppm, from about 1400 ppm to about 3400 ppm, from about 1400 ppm to about 2400 ppm, from about 1400 ppm to about 2000 ppm, from about 1400 ppm to about 1700 ppm, from about 1700 ppm to about 3400 ppm, from about 1700 ppm to about 2400 ppm, from about 1700 ppm to about 2000 ppm, from about 2000 ppm to about 3400 ppm, from about 2000 ppm to about 2400 ppm, or from about 2400 ppm to about 3400 ppm.
  • the oil described herein comprises less than about 3400 ppm tocopherol, less than about 2400 ppm tocopherol, less than about 2000 ppm tocopherol, less than about 1700 ppm tocopherol, less than about 1400 ppm tocopherol, or less than about 900 ppm tocopherol.
  • the oil described herein comprises at least about 900 ppm tocopherol, at least about 1400 ppm tocopherol, at least about 1700 ppm tocopherol, at least about 2000 ppm tocopherol, at least about 2400 ppm tocopherol, or at least about 3400 ppm tocopherol.
  • the oil described herein comprises about 1400 ppm tocopherol, about 1700 ppm tocopherol, or about 2400 ppm tocopherol.
  • addition tocopherol when used herein includes isomers and derivatives of tocopherol that are added to an oil described herein.
  • Addition tocopherols include, for example, ⁇ -tocopherol, dl- ⁇ -tocopherol, d- ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, D- ⁇ -tocopherol, ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, and/or mixtures thereof.
  • the oil described herein may comprise an addition tocopherol ranging from about 0 ppm to about 2500 ppm, from about 0 ppm to about 1500 ppm, from about 0 ppm to about 900 ppm from about 0 ppm to about 800 ppm, from about 0 ppm to about 500 ppm, from about 50 ppm to about 5000 ppm, from about 500 ppm to about 3500 ppm, from about 500 ppm to about 2500 ppm, from about 500 ppm to about 1500 ppm, from about 500 ppm to about 900 ppm, from about 500 ppm to about 800 ppm, from about 300 ppm to about 700 ppm, from about 800 ppm to about 2500 ppm, from about 800 ppm to about 1500 ppm, from about 800 ppm to about 900 ppm, from about 900 ppm to about 2500 ppm, from about 900 ppm to about 1500 ppm, from about
  • the oil described herein comprises less than about 2500 ppm addition tocopherol, less than about 1500 ppm addition tocopherol, less than about 900 ppm addition tocopherol, less than about 800 ppm addition tocopherol, or less than about 500 ppm addition tocopherol. In another embodiment, the oil described herein comprises about 500 ppm addition tocopherol, about 800 ppm addition tocopherol, or about 1500 ppm addition tocopherol.
  • the tocopherol is chosen from ⁇ -tocopherol, dl- ⁇ -tocopherol, d- ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, D- ⁇ -tocopherol, ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, and/or mixtures thereof.
  • mixtures of isomers and derivatives of addition tocopherols including, for example, mixtures of dl- ⁇ -tocopherol, d- ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocopherol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, ⁇ -tocotrienol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, and D- ⁇ -tocopherol.
  • the mixed tocopherol is a mixture of D- ⁇ -tocopherol, D- ⁇ -tocopherol, D- ⁇ -tocopherol, and D- ⁇ -tocopherol. In another embodiment, the mixed tocopherol is a mixture of from about 9 to about 20% D- ⁇ -tocopherol, from about 1 to about 4% D- ⁇ -tocopherol, from about 50 to about 65% D- ⁇ -tocopherol, and from about 20 to about 35% D- ⁇ -tocopherol. In yet another embodiment, the mixed tocopherol is a natural mixed tocopherol.
  • the oil described herein comprises mixed tocopherol in an amount ranging from about 900 ppm to about 3400 ppm, from about 900 ppm to about 2400 ppm, from about 900 ppm to about 2000 ppm, from about 900 ppm to about 1700 ppm, from about 900 ppm to about 1400 ppm, from about 1400 ppm to about 3400 ppm, from about 1400 ppm to about 2400 ppm, from about 1400 ppm to about 2000 ppm, from about 1400 ppm to about 1700 ppm, from about 1700 ppm to about 3400 ppm, from about 1700 ppm to about 2400 ppm, from about 1700 ppm to about 2000 ppm, from about 2000 ppm to about 3400 ppm, from about 2000 ppm to about 2400 ppm, or from about 2400 ppm to about 3400 ppm.
  • the oil described herein comprises less than about 3400 ppm mixed tocopherol, less than about 2400 ppm mixed tocopherol, less than about 2000 ppm mixed tocopherol, less than about 1700 ppm mixed tocopherol, less than about 1400 ppm mixed tocopherol, or less than about 900 ppm mixed tocopherol. In one embodiment, the oil described herein comprises about 1400 ppm mixed tocopherol, about 1700 ppm mixed tocopherol, or about 2400 ppm mixed tocopherol.
  • oils described herein may further contain natural tocopherols, in the form of, for example, tocotrienols that the microorganism produces during fermentation and which is ultimately contained in the crude oil.
  • the amount of natural tocopherols contained in the crude oil can range from, for example, about 25 ppm to about 500 ppm tocotrienols.
  • green tea extract includes, for example, green tea extracts containing, for example, polyphenolic compounds (i.e., catechins).
  • polyphenolic compounds i.e., catechins
  • examples of polyphenolic compounds that can be present in green tea extracts include epigallocatechin gallate 25 (EGCG), epigallocatechin (EGC), epicatechin gallate (ECG), epicatechin (EC), and mixture thereof.
  • the green tea extract contains at least one polyphenolic compound in an amount of from about 1% to about 90%, from about 5% to about 85%, from about 10% to about 80%, from about 15% to about 75%, from about 20% to about 70%, from about 25% to about 65%, from about 30% to about 60%, from about 35% to about 55%, or from about 40% to about 50% by weight of the green tea extract, exclusive of any carriers.
  • flavoring reagent includes, for example, rosemary extract and vanilla.
  • the rosemary extract may be extracted from, for example, Rosmarinus officinalis .
  • the rosemary extract may, for example, be organic.
  • the rosemary extract may be obtained by drying leaves of rosemary, which belongs to the Perilla family, pulverizing the dried leaves, and subjecting the resultant pulverized material to extraction with water, hot water, hexane, ethanol, acetone, ethyl acetate, or a mixture of any of these solvents.
  • rosemary extract examples include caffeic acid, carnosol, carnosic acid, methoxy carnosic acid, rosmarinic acid, rosmanol, rosmaridiphenol, rosmaridiquinone, and/or mixtures thereof.
  • Numerous rosemary extracts are also available commercially, and any one or more can be used in the present invention.
  • Suitable rosemary extracts are commercially available from, for example, Kalsec (Kalamazoo, Mich., USA) under the trade name of Herbalox®; Vitiva (Markovci, Slovenia) under the trade name Inolens®; Naturex (Avignon, France) under the trade name StabileEnhance®; and Ecom Food Industries Corporation (Ontario, Canada) under the product code NR 3401.
  • the flavoring reagent is rosemary extract.
  • the rosemary extract is extracted from Rosmarinus officinalis.
  • the oil described herein may comprise rosemary extract in an amount ranging from about 0 ppm to about 5000 ppm, from about 0 ppm to about 3500 ppm, from about 0 ppm to about 3000 ppm, from about 0 ppm to about 2000 ppm, from about 0 ppm to about 750 ppm, from about 0 ppm to about 500 ppm, from about 50 ppm to about 5000 ppm, from 50 ppm to about 7,500 ppm, from about 50 ppm to about 10,000 ppm, from about 500 ppm to about 5000 ppm, from about 500 ppm to about 4000 ppm, from about 500 to about 3500 ppm, from about 500 ppm to about 3000 ppm, from about 500 ppm to about 2000 ppm, from about 500 ppm to about 750 ppm, from about 750 ppm to about 5000 ppm, from about 750 ppm to about 3500 ppm, from about
  • the oil described herein comprises less than about 5000 ppm rosemary extract, less than about 3500 ppm rosemary extract, less than about 3000 ppm rosemary extract, less than about 200 ppm rosemary extract, less than about 750 ppm rosemary extract, or less than about 500 ppm rosemary extract. In yet still another embodiment, the oil described herein comprises about 5000 ppm rosemary extract, about 3500 ppm rosemary extract, about 3000 ppm rosemary extract, or about 2000 ppm rosemary extract.
  • the oil described herein may comprise about 3000 ppm rosemary extract, about 1700 mixed tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is essentially free of lecithin.
  • the oil described herein may comprise about 3000 ppm rosemary extract, about 800 ppm addition tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is essentially free of lecithin.
  • the oil described herein may comprise about 3500 ppm rosemary extract and about 1400 ppm mixed tocopherol, with the proviso that the oil is essentially free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 3500 ppm rosemary extract and about 500 ppm addition tocopherol, with the proviso that the oil is essentially free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 2000 ppm rosemary extract and about 2400 ppm mixed tocopherol, with the proviso that the oil is essentially free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 2000 ppm rosemary extract and about 1500 ppm addition tocopherol, with the proviso that the oil is essentially free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 5000 ppm rosemary extract, about 1700 mixed tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is essentially free of lecithin.
  • the oil described herein may comprise about 5000 ppm rosemary extract, about 800 ppm addition tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is essentially free of lecithin.
  • the oil described herein may comprise about 3000 ppm rosemary extract, about 1700 mixed tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is free of lecithin.
  • the oil described herein may comprise about 3000 ppm rosemary extract, about 800 ppm addition tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is free of lecithin.
  • the oil described herein may comprise about 3500 ppm rosemary extract and about 1400 ppm mixed tocopherol, with the proviso that the oil is free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 3500 ppm rosemary extract and about 500 ppm addition tocopherol, with the proviso that the oil is free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 2000 ppm rosemary extract and about 2400 ppm mixed tocopherol, with the proviso that the oil is free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 2000 ppm rosemary extract and about 1500 ppm addition tocopherol, with the proviso that the oil is free of lecithin and ascorbyl palmitate.
  • the oil described herein may comprise about 5000 ppm rosemary extract, about 1700 mixed tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is free of lecithin.
  • the oil described herein may comprise about 5000 ppm rosemary extract, about 800 ppm addition tocopherol, and about 250 ppm ascorbyl palmitate, with the proviso that the oil is free of lecithin.
  • One embodiment is directed to an oil comprising at least about 30%, by weight of fatty acid content in the oil, of at least one polyunsaturated fatty acid having at least 4 carbon-carbon double bonds; at least one first antioxidant; and means for improving oxidative stability.
  • the p-Anisidine value (p-AV) is determined in accordance with AOCS Official Method Cd 18-90.
  • the oil described herein has a p-AV of less than about 40; less than about 30; or less than about 20.
  • the peroxide value (PV) is determined in accordance with the AOCS Official Method Cd 8-53.
  • the oil described herein has a PV less than about 20 meq/kg; less than about 10 meq/kg; or less than about 5 meq/kg.
  • the Rancimat values are determined by the standard test for oil stability, using a rancimat apparatus operated at 90° C., with airflow set at 10 L/hour (AOCS Cd 12b-92).
  • the oil described herein has a RANCIMAT value of less than or equal to about 3, to about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19 at suitable storage.
  • rancimat value is determined after about 3 months, about 5 months, about 6 months, about 7 months, or 8 months of suitable storage. A person of skill in the understand would understand at what conditions to store the oils described herein.
  • the oil described herein is stored at room temperature (25° C.) in Nylon/Foil/PE Low Density Polyethylene bags packaged with vacuum then heat sealed under N 2 (Heritage Packaging) or epoxy-phenolic lined aluminium containers (Elemental Container Inc.)
  • the marine/fishy aroma (smell) and marine/fishy aromatics (taste) sensory values were determined according to the method as described in Sensory Evaluation Techniques. Meilgaard et al., CRC Press; 4 edition (Dec. 13, 2006).
  • a panel of 8-18 experienced people taste (or smell) a sample. Each of these people determine the value of the sample. Afterwards all the values are averages arithmetically and the result is rounded up or down to the next number.
  • a value of ⁇ 1.5 fishy/marine aroma and ⁇ 2.5 fishy/marine aromatics is expected to be perceivable by the general population.
  • the oil described herein has a fishy aroma sensory value of less than 1.5 after about 3 months, after about 4 months, after about 5 months, after about 6 months, after about 7 months, after about 8 months, or after about 9 months of suitable storage. In another embodiment, the oil described herein has a fishy aroma value of less than 1.5 at a time period chosen from 0-3 months, at 0-4 months, at 0-5 months, at 0-6 months, at 0-7 months, at 0-8 months, or at 0-9 months.
  • the suitable storage is performed at room temperature (25° C.) in Nylon/Foil/PE Low Density Polyethylene bags packaged with vacuum then heat sealed under N 2 or epoxy-phenolic lined aluminium containers.
  • the oil described herein has a fishy aromatics sensory value of less than 2.5 after about 3 months, after about 4 months, after about 5 months, after about 6 months, after about 7 months, after about 8 months, or after about 9 months of suitable storage.
  • the oil described herein has a fishy aroma value of less than 1.5 at a time period chosen from 0-3 months, at 0-4 months, at 0-5 months, at 0-6 months, at 0-7 months, at 0-8 months, or at 0-9 months.
  • the suitable storage is performed at room temperature (25° C.) in Nylon/Foil/PE Low Density Polyethylene bags packaged with vacuum then heat sealed under N 2 or epoxy-phenolic lined aluminium containers.
  • the at least one C 18-22 PUFA having 4 carbon-carbon double bonds is chosen from docasoahexaenoic acid (“DHA”), eicosapentaenoic acid (“EPA”), arachidonic acid (“ARA”), omega-3 docosapentaenoic acid (“DPA n-3”), and omega-6 docosapentaenoic acid (“DPA n-6”).
  • DHA docasoahexaenoic acid
  • EPA eicosapentaenoic acid
  • ARA arachidonic acid
  • DPA n-3 omega-3 docosapentaenoic acid
  • DPA n-6 omega-6 docosapentaenoic acid
  • the oil comprises omega-3 PUFAs.
  • the omega-3 PUFAs are chosen from DHA, EPA, DPAn-3, and mixtures thereof.
  • the oil is characterized by at least one of the following fatty acids (or esters thereof), expressed as wt % of the total fatty acid content of the oil.
  • the embodiments described herein may further comprise about 3% or less of other fatty acids or esters thereof.
  • the oil described herein comprises at least about 30 wt. %, at least about 35 wt. %, at least about 40 wt. %, at least about 45 wt. %, or at least about 50 wt. % of one or more polyunsaturated fatty acids having at least 4 double bonds.
  • the oil described herein comprises from about 30 wt. % to about 60 wt. %, from about 30 wt. % to about 50 wt. %, from about 30 wt. % to about 40 wt. %, from about 40 wt. % to about 60 wt. %, or from about 40 wt. % to about 50 wt. % of one or more polyunsaturated fatty acid having at least 4 double bonds.
  • the oil described herein comprises at least about 30 wt. %, at least about 35 wt. %, at least about 40 wt. %, at least about 45 wt. %, or at least about 50 wt. % of DHA. In another embodiment, the oil described herein comprises from about 30 wt. % to about 60 wt. %, from about 30 wt. % to about 50 wt. %, from about 30 wt. % to about 40 wt. %, from about 40 wt. % to about 60 wt. %, or from about 40 wt. % to about 50 wt. % of DHA.
  • the oil described herein comprises at least about 35 wt. % of DHA+EPA, at least about 40 wt. %, at least about 45 wt. %, at least about 50 wt. % of DHA+EPA, wherein said oil comprises less than about 80 wt. %, less than about 70 wt. %, or less than about 60 wt. % of DHA+EPA.
  • the oil described herein comprises at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50%, by weight of the total fatty acid content of the oil, of at least one polyunsaturated fatty acid having at least 4 double bonds.
  • the oil described herein comprises from about 30% to about 60%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, or from about 40% to about 50%, by weight of the total fatty acid content of the oil, of at least one polyunsaturated fatty acid having at least 4 double bonds.
  • the oil described herein comprises at least about 30%, at least about 35%, at least about 40%, at least about 45%, or at least about 50%, by weight of the total fatty acid content of the oil, of DHA. In another embodiment, the oil described herein comprises from about 30% to about 60%, from about 30% to about 50%, from about 30% to about 40%, from about 40% to about 60%, or from about 40% to about 50%, by weight of the total fatty acid content of the oil, of DHA.
  • the oil comprises less than about 80%, less than about 70%, or less than about 60%, by weight totally fatty acid, of at least one C 18-22 PUFA having 4 carbon-carbon double bonds.
  • the oil described herein comprises from about 200 mg DHA/g oil, from about 300 mg DHA/g oil, from about 350 mg DHA/g oil, from about 400 mg DHA/g oil, or from about 500 mg DHA/g oil. In another embodiment, the oil described herein comprises from about 120 mg EPA/g oil or from about 130 mg EPA/g oil.
  • the oil described herein comprises from about 200 mg DHA/g oil to about 600 mg DHA/g oil, from about 200 mg DHA/g oil to about 500 mg DHA/g oil, from about 200 mg DHA/g oil to about 400 mg DHA/g oil, from about 300 mg DHA/g oil to about 600 mg DHA/g oil, from about 300 mg DHA/g oil to about 500 mg DHA/g oil, or from about 300 mg DHA/g oil to about 400 mg DHA/g oil.
  • the oil described herein comprises from about 100 mg EPA/g oil to about 250 mg EPA/g oil.
  • the oil described herein comprises from about 400 mg DHA+EPA/g oil or from about 500 mg DHA+EPA/g oil.
  • the oil described herein may comprise about 2% or less of ARA, by weight of the total fatty acid content of the oil. In a further embodiment, the oil described herein comprises about 3% or less of EPA, by weight of the total fatty acid content of the oil. In a still further embodiment, the oil described herein comprise about 18% or less or about 12% to about 18%, by weight of the total fatty acid content of the oil, of DPA n-6. In yet an even further embodiment, the oil described herein may comprise about 10% or less, by weight of the total fatty acid content of the oil, of other fatty acids.
  • the oil is substantially free of EPA.
  • the term “substantially free of EPA” may refer to an oil in which EPA is less than about 3%, by weight of the total fatty acid content of the oil.
  • the oil 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.
  • the oil has no detectable amount of EPA using techniques known in the art. In some embodiments, the oil has no EPA.
  • the oil can also be substantially free of ARA.
  • the ARA is less than about 3% by weight of the total fatty acid content of the oil.
  • ARA comprises less than about 2% by weight of the total fatty acid content of the oil, less than about 1% by weight of the total fatty acid content of the oil, less than about 0.5% by weight of the total fatty acid content of the oil, less than about 0.2% by weight of the total fatty acid content of the oil, or less than about 0.01% by weight of the total fatty acid content of the oil.
  • the oil has no detectable amount of ARA.
  • the oil described herein may comprise about 2 wt. % or less of ARA. In a further embodiment, the oil described herein comprises about 3 wt. % or less of EPA. In a still further embodiment, the oil described herein comprise about 18 wt. % or less or about 12 wt. % to about 18 wt. % of DPA n-6. In yet an even further embodiment, the oil described herein may comprise about 10 wt. % or less of other fatty acids.
  • the oil is substantially free of EPA.
  • the term “substantially free of EPA” may refer to an oil in which EPA is less than about 3 wt. %.
  • the oil comprises, less than about 2 wt. % EPA less than about 1 wt. % EPA, less than about 0.5 wt. % EPA, less than about 0.2 wt. % EPA, or less than about 0.01 wt. % EPA.
  • the oil has no detectable amount of EPA using techniques known in the art. In some embodiments, the oil has no EPA.
  • the oil can also be substantially free of ARA. In some embodiments, the oil comprises less than about 3 wt. % ARA. In some embodiments, the oil comprises less than 2 wt. % ARA, less than about 1 wt. %, less than about 0.5 wt. %, less than about 0.2 wt. %, or less than about 0.01 wt. % ARA. In some embodiments, the oil has no detectable amount of ARA.
  • the oil describe herein can be used in any application, where such oils are needed.
  • the oil can be used in, for example, food products (including beverages and dietary supplements), animal feed, and/or personal care products. These products can be in, for example, any form, such as, a liquid, emulsion, gel, and/or solid. These products can be ready to use (ready to consume) products as well as products which need to be further processed (for example by dilution, dissolving, heating, etc).
  • the oil disclosed herein is an edible oil.
  • the oil disclosed herein is an edible oil that is used in food products.
  • Exemplary food products include, but are not limited to nutritional bars, dietary supplements, granola bars, baked goods (e.g., breads, rolls, cookies, crackers, fruit pies, or cakes), pastas, condiments, salad dressings, soup mixes, snack foods, processed fruit juices, sauces, gravies, syrups, beverages, dry beverage powders, and jams or jellies.
  • baked goods e.g., breads, rolls, cookies, crackers, fruit pies, or cakes
  • pastas e.g., breads, rolls, cookies, crackers, fruit pies, or cakes
  • pastas e.g., condiments, salad dressings
  • soup mixes e.g., snack foods, processed fruit juices, sauces, gravies, syrups, beverages, dry beverage powders, and jams or jellies.
  • the oil described herein is organic.
  • organic includes, for example, the standards set by the U.S. Department of Agriculture (USDA) and the European Union (EU) for including this term in food product labeling.
  • the EU standards are set forth, for example, in Regulation EC 834/2007, and in the US the USDA standards are set forth, for example, in the National Organic Program Regulation at 7 C.F.R., Part 205.
  • the oil described here is natural.
  • naturally includes, for example, using this term in food product labeling associated with food products that do not contain added color, artificial flavors, or synthetic substances.
  • the at least one C 18-22 PUFA having 4 carbon-carbon double bonds can be obtained from various sources including, for example, aquatic animals, such as, fish, marine mammals, and crustaceans (such as hill and other euphausids); animal sources including, for example, animal tissues that include for example brain, liver, and eyes and animal products that include, for example, eggs and milk; microalgae; plant; and/or seed.
  • the oil is obtained from fish, microalgae, plant or seed.
  • the at least one C 18-22 PUFA having 4 carbon-carbon double bonds is obtained from microalgae.
  • the microalgae is from the order Thraustochytriales.
  • the order Thraustochytriales includes, for example, the genera Thraustochytrium (species include arudimentale, aureum, benthicola, globosum, kinnei, motivum, multirudimentale, pachydermum, proliferum, roseum, striatum ), the genera Schizochytrium (species include aggregatum, limnaceum, mangrovei, minutum, octosporum ), the genera Ulkenia (species include amoeboidea, kerguelensis, minuta, profunda, radiate, sailens, sarkariana, schizochytrops, visurgensis, yorkensis ), the genera Aurantiacochytrium ; the genera Oblongichytrium , the genera Thraus
  • microalgae is Thraustochytrium sp.
  • microalgae is Schizochytrium sp.
  • microalgae is chosen from Thraustochytrium sp. and Schizochytrium sp.
  • the at least one C 18-22 PUFA having 4 carbon-carbon double bonds is obtained from plants, grown either in culture fermentation or in crop plants, including, for example, cereals (such as maize, barley, wheat, rice, sorghum, pearl millet, corn, rye and oats); beans; soybeans; peppers; lettuce; peas; Brassica species, such as, cabbage, broccoli, cauliflower, brussel sprouts, rapeseed, and radish; carrot; beets; eggplant; spinach; cucumber; squash; melons; cantaloupe; sunflowers; safflower; canola; flax; peanut; mustard; rapeseed; chickpea; lentil; white clover; olive; palm; borage; evening primrose; linseed; and tobacco.
  • cereals such as maize, barley, wheat, rice, sorghum, pearl millet, corn, rye and oats
  • beans soybeans
  • peppers lettuce
  • peas Brassica species, such as
  • the oil described herein is a crude oil. In another embodiment, the oil described herein is a refined oil. In yet a further embodiment, the oil described herein is a final oil.
  • a “crude oil” is an oil that is extracted from the biomass of a microorganism without further processing.
  • a “refined oil” is an oil that is obtained by treating a crude oil with standard processing of refining, bleaching, and/or deodorizing. See, e.g., U.S. Pat. No. 5,130,242.
  • a “final oil” is a refined oil that is further blended with a vegetable oil.
  • a final oil is a refined oil that has been blended with a vegetable oil chosen from medium chain triglycerides (MCTs), canola oil, palm oil, and sunflower oil.
  • MCTs medium chain triglycerides
  • the sunflower oil is high oleic sunflower oil.
  • the sunflower oil is organic.
  • the high oleic sunflower oil is organic.
  • One embodiment is directed to a method for improving the oxidative stability of an oil, comprising adding an effective amount of at least one first antioxidant to an oil comprising at least 30%, by weight of fatty acid content in the oil, of at least one polyunsaturated fatty acid having at least 4 double bonds
  • the microbial oils described herein can be recovered from microalgae by any suitable means known to those in the art.
  • the oils can be recovered by extracting with techniques, such as those described in, for example, International Pub. Nos. WO 2001/053512, WO 2001/051598, WO 2001/076715, and WO 2001/076385; U.S. Pub. Nos. 2007/0004678 and 2005/012739; and U.S. Pat. No. 6,399,803.
  • Processes for the enzyme treatment of biomass for the recovery of lipids are disclosed in International Pub. No. WO 2003/09628; U.S. Pub. No. 2005/0170479; EP Pat. Pub. 0776356 and U.S. Pat. No. 5,928,696.
  • the oil described herein is obtained via the following steps: generating biomass by fermenting microalgae capable of producing oil that contains at least one C 18-22 PUFA having 4 carbon-carbon double bonds; harvesting the biomass; spray drying the biomass; extracting oil from the biomass; refining the oil (to remove free fatty acids and phospholipids); bleaching the oil (to remove any remaining polar compounds and pro-oxidant metals, and to break down lipid oxidation products); chill filtering the oil (to remove any remaining insoluble fats, waxes, and solids); deodorizing the oil (optionally under vacuum and in, for example, a packed column, counter current steam stripping deodorizer); adding an antioxidant to the oil; and any combinations thereof.
  • the culture is harvested by centrifugation then pasteurized and spray dried.
  • the dried biomass is flushed with nitrogen and packaged before being stored frozen at ⁇ 20° C.
  • the oil is extracted from the dried biomass by mixing the biomass with n-hexane or isohexane in a batch process which disrupts the cells and allows the oil and cellular debris to be separated.
  • the solvent is then removed.
  • the oil described herein is stored at room temperature (25° C.) in Nylon/Foil/PE Low Density Polyethylene bags packaged with vacuum then heat sealed under N 2 on a Model AGV Multivac. (Multivac Sepp Haggenmüller GmbH & Co. KG).
  • the oils described herein are stored at room temperature (25° C.) in epoxy-phenolic lined aluminium containers.
  • the oil can comprise further ingredients, which can be useful for the final product and/or for the production process of the final product.
  • further ingredients can include, for example, colorants, fragrances, fillers, flavors, non-lecithin emulsifiers, stabilizers, and other lipophilic materials.
  • the rosemary extract used in examples 3-15 is available from, for example, Ecom Foods Industries Corporation (Ontario, Canada).
  • the high oleic sunflower oil used in examples 3-15 is available, for example, from Humko oils (Memphis, Tenn.) under the trade name TriSun®.
  • TAP1010 Sun is available from Vitablend (Wolvega, the Netherlands).
  • the soy lecithin used in the examples below is available, for example, from Archer Daniels Midland Co. (Decatur, Ill.) under the trade name Yelkin® Gold.
  • the ascorbyl palmitate used in the examples below is available, for example, from DSM, Nutritional Products (Basel Switzerland).
  • PTA-10208 Shizochytrium sp. deposited under ATCC Accession No. PTA-10208 (hereinafter referred to as “PTA-10208”) was grown via individual fermentation runs, as described below. Typical media and cultivation conditions are set forth in Table 1.
  • PTA-10208 produced a dry cell weight of 95 g/L after 200 hours of culture in a 10 L fermentor volume.
  • the lipid yield was 53.7 g/L; the omega-3 yield was 37 g/L; the EPA yield was 14.3 g/L; and the DHA yield was 21 g/L.
  • the fatty acid content was 57% by weight; the EPA content was 27.7% of FAME; and the DHA content was 39.1% of FAME.
  • the lipid productivity was 6.4 g/L/day, and the omega-3 productivity was 4.4 g/L/day under these conditions, with 1.7 g/L/day EPA productivity and 2.5 g/L/day DHA productivity.
  • PTA-10208 produced a dry cell weight of 56 g/L after 139 hours of culture in a 10 L fermentor volume.
  • the lipid yield was 53 g/L; the omega-3 yield was 34 g/L; the EPA yield was 11.5 g/L; and the DHA yield was 22 g/L.
  • the fatty acid content was 58% by weight; the EPA content was 21.7% of FAME; and the DHA content was 41.7% of FAME.
  • the lipid productivity was 9.2 g/L/day, and the omega-3 productivity was 5.9 g/L/day under these conditions, with 2 g/L/day EPA productivity and 3.8 g/L/day DHA productivity.
  • PTA-10208 produced a dry cell weight of 93.8 g/L after 167 hours of culture in a 2000 L fermentor volume.
  • the lipid yield was 47.2 g/L; the omega-3 yield was 33.1 g/L; the EPA yield was 10.5 g/L; and the DHA yield was 20.4 g/L.
  • the fatty acid content was 50.6% by weight; the EPA content was 23% of FAME; and the DHA content was 42.6% of FAME.
  • the lipid productivity was 6.8 g/L/day, and the omega-3 productivity was 4.7 g/L/day under these conditions, with 1.5 g/L/day EPA productivity and 2.9 g/L/day DHA productivity.
  • PTA-10208 produced a dry cell weight of 105 g/L after 168 hours of culture in a 2000 L fermentor volume.
  • the lipid yield was 46.4 g/L; the omega-3 yield was 33 g/L; the EPA yield was 10.7 g/L; and the DHA yield was 20.3 g/L.
  • the fatty acid content was 43.9% by weight; the EPA content was 24% of FAME; and the DHA content was 43.7% of FAME.
  • the lipid productivity was 6.6 g/L/day, and the omega-3 productivity was 4.7 g/L/day under these conditions, with 1.5 g/L/day EPA productivity and 2.9 g/L/day DHA productivity.
  • PTA-10208 produced a dry cell weight of 64.8 g/L after 168 hours of culture in a 2000 L fermentor volume.
  • the lipid yield was 38.7 g/L; the omega-3 yield was 29.9 g/L; the EPA yield was 8.5 g/L; and the DHA yield was 16.7 g/L.
  • the fatty acid content was 59.6% by weight; the EPA content was 23% of FAME; and the DHA content was 42.3% of FAME.
  • the lipid productivity was 5.53 g/L/day, and the omega-3 productivity was 3.8 g/L/day under these conditions, with 1.2 g/L/day EPA productivity and 2.3 g/L/day DHA productivity.
  • Example 2 Two samples of the biomass produced in accordance with Example 1 (PTA-10208 Sample #1 and PTA-10208 Sample #2) were analyzed for total crude oil content by solvent extraction, lipid classes were determined by high performance liquid chromatography/evaporative light scattering detection (HPLC/ELSD), triacylglycerol (TAG) was analyzed by HPLC/mass spectrometry (HPLC/MS), and fatty acid (FA) profiles were determined by gas chromatography with flame ionization detection (GC-FID).
  • HPLC/ELSD high performance liquid chromatography/evaporative light scattering detection
  • TAG triacylglycerol
  • HPLC/MS HPLC/mass spectrometry
  • FA fatty acid
  • the crude lipid content of each freeze dried biomass was determined using solvent grinding with hexane and compared to the sum of FAME (mg/g) generated by direct transesterification, and the resultant fatty acid methyl esters (FAME) were quantified by GC/FID analysis.
  • FAs in the extracted crude lipid were also quantified by transesterification and quantified using GC/FID analysis of the resultant FAME.
  • the weight percent of all neutral lipids (NL) and free fatty acids (FFA) were determined in the extracted crude lipid using normal phase HPLC with ELSD and atmospheric pressure chemical ionization-MS (APCI-MS) identification.
  • the method separates and quantifies sterol esters (SE), TAG, FFAs, 1,3-diacylglycerols (1,3-DAG), sterols, 1,2-diacylglycerols (1,2-DAG), and monoacylglycerols (MAG). Results are shown in Tables 2 and 3.
  • TAG and phospholipids were isolated from the extracted crude oil (PTA-10208 Sample #1 and PTA-10208 Sample #2). TAG was isolated using low pressure flash chromatography and PL was isolated using solid phase extraction (SPE). The identity of each isolated fraction was confirmed by thin layer chromatography (TLC). The fatty acid profiles of the isolated TAG and PL fractions were determined following direct transesterification using GC-FID as FAME. Results are shown in Table 4.
  • Crude Oil Extraction was extracted from samples of freeze-dried biomass using solvent grinding. For example, approximately 3 grams of biomass was weighed into a Swedish tube. Three ball bearings and 30 mL of hexane were added to the Swedish tube, which was sealed with a neoprene stopper and placed in a shaker for 2 hours. The resultant slurry was filtered using a Buchner funnel and Whatman filter paper. The filtered liquid was collected, the solvent removed under vacuum, and the amount of remaining crude lipid determined gravimetrically.
  • Fatty Acid Analysis The samples of biomass, extracted crude lipid, and isolated lipid classes were analyzed for fatty acid composition as FAME. Briefly, freeze-dried biomass and isolated lipid classes were weighed directly into a screw cap test tubes, while samples of the crude oil were dissolved in hexane to give a concentration of approximately 2 mg/mL. Toluene, containing internal standard, and 1.5 N HCl in methanol was added to each tube. The tubes were vortexed, then capped and heated to 100° C. for 2 hours. The tubes were allowed to cool, and saturated NaCl in water was added. The tubes were vortexed again and centrifuged to allow the layers to separate.
  • Solid Phase Extraction PL fractions were separated from the crude lipid by solid phase extraction (SPE) using 2 g aminopropyl cartridges (Biotage, Uppsala, Sweden) placed in a Vac Elut apparatus (Varian Inc, Palo Alto, USA). The cartridge was conditioned with 15 mL of hexane, and ⁇ 60 mg of each sample was dissolved in 1 mL CHCl 3 and applied to the cartridge. The column was washed with 15 mL of 2:1 CHCl 3 :isopropyl alcohol to elute all the neutral lipids, which was discarded. The fatty acids were then eluted with 15 mL of 2% acetic acid (HOAc) in ether, which was discarded. The PL portion was eluted with 15 mL of 6:1 Methanol:Chloroform, which was collected, dried under nitrogen, and weighed.
  • SPE solid phase extraction
  • Flash Chromatography Flash chromatography was used to separate the lipid classes present in the crude oil. Approximately 200 mg of crude oil dissolved in hexane was injected onto the head of the column. The chromatography system utilized Silica Gel 60 (EMD Chemical, Gibbstown, N.J.) with mobile phase composed of Petroleum Ether and Ethyl Acetate at 5 mL/min (Tables 6-7) or 3 mL/min (Tables 8-13). A step gradient was used to selectively elute each lipid class from the column. The mobile phase gradient started from 100% petroleum ether and finished with 50% ethyl acetate.
  • Fractions were collected in 10 mL test tubes using a Gilson FC 204 large-bed fraction collector (Gilson, Inc., Middleton, Wis.). Each tube was analyzed by thin layer chromatography (TLC) and the tubes containing individual lipid classes (as judged by single spots on TLC plate with expected retention factor (Rf)) were pooled, concentrated to dryness, and weighed. The total fraction content was then determined gravimetrically.
  • TLC thin layer chromatography
  • TAG and PL fractions were analyzed for fatty acid composition as fatty acid methyl esters (FAME).
  • FAME fatty acid methyl esters
  • the TAG fractions were dissolved in hexane to give a concentration of approximately 1-2 mg/mL. 1 mL aliquots of the solutions were concentrated to dryness under nitrogen. Toluene, containing internal standard, and 1.5 N HCl in methanol was added to each tube. The tubes were vortexed, then capped and heated to 100° C. for 2 hours. Internal standard and HCl methanol were added directly to the tubes containing the PL fraction and heated. The tubes were allowed to cool, and saturated NaCl in water was added.
  • FAME fatty acid methyl esters
  • the tubes were vortexed again and centrifuged to allow the layers to separate. A portion of the organic layer was then placed in a GC vial and analyzed by GC-FID. FAMEs were quantified using a 3-point calibration curve generated using Nu-Check-Prep GLC 502B Reference Standard (NuCheck, Elysian, Minn.). Fatty acids present in the extract were expressed as mg/g and as a % of FAME.
  • the fatty acid profile of the biomass and extracted crude lipid for PTA-10208 Sample #1 was determined using GC/FID.
  • FAs in the biomass were transesterified in situ by weighing 28.6 mg of biomass directly into a FAME tube, while a sample of the extracted crude lipid was prepared by weighing 55.0 mg of crude lipid into a 50 mL volumetric flask and transferring 1 ml to a separate FAME tube.
  • the estimated crude lipid content of the biomass was determined to be 53.2% (as SUM of FAME) using GC with FID detection, while 52.0% (wt/wt) lipid was extracted from the dry biomass, giving a 97.8% recovery of total lipid.
  • the crude lipid was determined to be 91.9% fatty acids (as SUM of FAME) using GC/FID.
  • the major fatty acids contained in the crude lipid were C16:0 (182.5 mg/g), C20:5 n-3 (186.8 mg/g), and C22:6 n-3 (423.1 mg/g).
  • the lipid class profile of the extracted crude lipid was determined by weighing 55.0 mg of crude lipid into a 50 mL volumetric flask and transferring an aliquot into an HPLC vial for HPLC/ELSD/MS analysis.
  • the crude lipid contained 0.2% sterol esters (SE), 95.1% TAG, 0.4% sterols, and 0.5% 1,2-diacylglycerol (DAG).
  • SE sterol esters
  • TAG 1,2-diacylglycerol
  • 5% of the TAG fraction included a peak that eluted directly after the TAG peak, but did not give a recognizable mass spectrum.
  • TAG Isolated TAG from this sample as determined by flash chromatography made up approximately 92.4% of the crude oil. PL was not detected by weight or TLC after SPE isolation.
  • the major fatty acids (>50 mg/g) contained in the TAG were C16:0 (189 mg/g), C20:5 n-3 (197 mg/g), and C22:6 n-3 (441 mg/g).
  • the fatty acid profile of the biomass and extracted crude lipid for PTA-10208 Sample #2 was determined using GC/FID.
  • FAs in the biomass were transesterified in situ by weighing 32.0 mg of biomass directly into a FAME tube, while a sample of the extracted crude lipid was prepared by weighing 60.1 mg of crude lipid into a 50 mL volumetric flask and transferring 1 ml to a separate FAME tube.
  • the estimated crude lipid content of the biomass was determined to be 52.4% (as SUM of FAME) using GC with FID detection, while 48.0% (wt/wt) lipid was extracted from the dry biomass, giving a 91.7% recovery of total lipid.
  • the crude lipid was determined to be 95.3% fatty acids (as SUM of FAME) using GC/FID.
  • the major fatty acids contained in the crude lipid were C16:0 (217.5 mg/g), C20:5 n-3 (169.3 mg/g), and C22:6 n-3 (444.1 mg/g).
  • the lipid class profile of the extracted crude lipid was determined by weighing 60.1 mg of crude lipid into a 50 mL volumetric flask and transferring an aliquot into an HPLC vial for HPLC/ELSD/MS analysis. According to the HPLC/ELSD/MS analysis, the crude lipid contained 0.2% SE, 95.7% TAG, 0.3% sterols, and 0.7% 1,2-DAG. 5.1% of the TAG fraction included a peak that eluted directly after the TAG peak, but did not give a recognizable mass spectrum.
  • TAG Isolated TAG from this sample made up approximately 93.9% of the crude oil. PL was not detected by weight or TLC after SPE isolation.
  • Example PTA-10208 #3 A sample of crude oil from the microorganism deposited under ATCC Accession No. PTA-10208 (Sample PTA-10208 #3) was analyzed using HPLC/ELSD/MS. A total of 98.38% of lipids were recovered, with the sterol ester (SE) fraction accounting for 0.32%, the TAG fraction accounting for 96.13%, the 1,3-diacylglycerol (DAG) fraction accounting for 0.22%, the 1,2-DAG fraction accounting for 0.78%, and the sterol fraction accounting for 0.93%.
  • SE sterol ester
  • DAG 1,3-diacylglycerol
  • sterol fraction accounting for 0.93%.
  • a crude oil can be obtained in accordance with the procedures set forth in examples 1 and 2.
  • a crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • a refined oil can be further blended with high oleic sunflower oil (“HOSO”) to achieve a final oil with a combined DHA+EPA content of at least about 400 mg/g oil.
  • HOSO high oleic sunflower oil
  • ingredients contained in the oil include 1200 ppm sunflower lecithin; 2000 ppm rosemary extract; 2000 ppm mixed tocopherols; and 300 ppm ascorbyl palmitate.
  • the 2000 ppm mixed tocopherol came from 1700 ppm of added tocopherols (includes 1100 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70, and from the mixed tocopherols contained in the 3000 ppm of TAP 1010 Sun that was added to the oil.
  • TAP1010 Sun provided 300 ppm ascorbyl palmitate, 300 ppm mixed tocopherols, and 1200 ppm sunflower lecithin.
  • a crude oil can be obtained in accordance with the procedures set forth in examples 1 and 2.
  • a crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • a refined oil can be further blended with HOSO to achieve a final oil with combined DHA+EPA content of at least about 500 mg/g oil.
  • Typical characteristics of the final oil according to this example are set forth in Table 14.
  • ingredients contained in the final oil include 1600 ppm sunflower lecithin; 2000 ppm Rosemary Extract; 2400 ppm mixed tocopherols; and 400 ppm ascorbyl palmitate.
  • the 2400 ppm mixed tocopherol came from 2000 ppm of added tocopherols (which includes 900 ppm that was added in upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70, and from the mixed tocopherols contained in the 4000 ppm of TAP 1010 Sun that was added to the oil.
  • TAP1010 Sun provided 400 ppm ascorbyl palmitate, 400 ppm mixed tocopherols, and 1600 ppm sunflower lecithin.
  • a crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • a refined oil can be further blended with HOSO to achieve a final oil with combined DHA+EPA content of at least about 500 mg/g oil.
  • the characteristics of the final oil according to this example are similar to the characteristics set forth in Table 14.
  • ingredients contained in the final oil include 3000 ppm Rosemary Extract; 1700 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 250 ppm ascorbyl palmitate.
  • the potency was obtained via the following protocol AOCS Ce 1b-89 (modified).
  • the marine/fishy aroma (smell) and marine/fishy aromatics (taste) sensory values were determined according to the method as described in Sensory Evaluation Techniques, Meilgaard et al., CRC Press; 4 edition (Dec. 13, 2006).
  • a panel of 8-18 experienced people tasted and/or smelled a sample of the final oils according to examples 4 and 5. Each of these people determined the value of the sample. Afterwards all the values were averaged arithmetically and the result was rounded up or down to the next number.
  • a value of ⁇ 1.5 fishy/marine aroma and ⁇ 2.5 fishy/marine aromatics is expected to be perceivable by the general population.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • a refined oil can be further blended with HOSO to achieve a final oil with a DHA content of at least about 350 mg/g oil.
  • Typical characteristics of final oil according to this example are set forth in Table 16.
  • ingredients contained in the final oil include 900 ppm mixed tocopherols that was added during upstream processing.
  • the mixed tocopherols are available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 900 ppm mixed tocopherols that was added during upstream processing; 400 ppm ascorbyl palmitate; and 2800 ppm soy lecithin.
  • the mixed tocopherols are available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 900 ppm mixed tocopherols that was added during upstream processing; 750 ppm ascorbyl palmitate; and 2800 ppm soy lecithin.
  • the mixed tocopherols are available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 3500 ppm Rosemary Extract; 1400 ppm mixed tocopherols; and 50 ppm ascorbyl palmitate.
  • the 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 2800 ppm Soy Lecithin; 3500 ppm Rosemary Extract; 1400 ppm mixed tocopherols; and 750 ppm ascorbyl palmitate.
  • the 1400 ppm mixed tocopherols includes 900 ppm that was added upstream, which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 2800 ppm Soy Lecithin; 3500 ppm Rosemary Extract; 3400 ppm mixed tocopherols; and 50 ppm ascorbyl palmitate.
  • the 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 2800 ppm Soy Lecithin; 3500 ppm Rosemary Extract; 3400 ppm mixed tocopherols; and 750 ppm ascorbyl palmitate.
  • the 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing) is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 500 ppm Rosemary Extract.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 500 ppm Rosemary Extract; 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 50 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 500 ppm Rosemary Extract; 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 750 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 500 ppm Rosemary Extract; 3400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 50 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 500 ppm Rosemary Extract; 3400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 750 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 2800 ppm soy lecithin; 2000 ppm Rosemary Extract; 2400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 400 ppm ascorbyl palmitate.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be further blended with HOSO to achieve final oil with DHA content of at least about 350 mg/g oil.
  • the characteristics of this oil are similar to the characteristics set forth in Table 16.
  • ingredients contained in the final oil include 3400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTML70; and 500 ppm Rosemary Extract.
  • Table 17 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • ingredients contained in the final oil include 1400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 3500 ppm Rosemary Extract.
  • Table 18 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • ingredients contained in the final oil include 3400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 3500 ppm Rosemary Extract.
  • Table 19 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • ingredients contained in the final oil include 2400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 2000 ppm Rosemary Extract.
  • Table 20 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • ingredients contained in the final oil include 2400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 0 ppm Rosemary Extract.
  • Table 21 summarizes the oil according to this example that does not contain ascorbyl palmitate, lecithin, or rosemary extract.
  • ingredients contained in the final oil include 900 ppm mixed tocopherols (that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 2000 ppm Rosemary Extract.
  • Table 22 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • ingredients contained in the final oil include 2400 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 2000 ppm Rosemary Extract.
  • Table 23 summarizes the oil according to this example that does not contain ascorbyl palmitate or lecithin.
  • the Rancimat values are determined by the standard test for oil stability, using a rancimat apparatus operated at 90° C., with airflow set at 10 L/hour (AOCS Cd 12b-92). The potency was obtained via the following protocol AOCS Ce 1b-89 (modified).
  • the marine/fishy aroma (smell) and marine/fishy aromatics (taste) sensory values were determined according to the method as described in Sensory Evaluation Techniques, Meilgaard et al., CRC Press; 4 edition (Dec. 13, 2006).
  • a panel of 8-18 experienced people tasted and/or smelled a sample a sample of the final oils according to examples 7a-m and 8a-g. Each of these people determined the value of the sample.
  • examples 7a-m and 8a-g were packaged in in Nylon/Foil/PE Low Density Polyethylene bags packaged with vacuum then heat sealed under N 2 on a Model AGV Multivac. (Multivac Sepp Haggenmüller GmbH & Co. KG) and stored at 25° C.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be further blended with HOSO to achieve a final oil with DHA content of at least about 350 mg/g oil.
  • Typical characteristics of the final oil according to this example are set forth in Table 25.
  • TAP1010 Sun provided 400 ppm ascorbyl palmitate, 400 ppm mixed tocopherols, and 1600 ppm sunflower lecithin.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be further blended with HOSO to achieve final oil with DHA content of at least about 400 mg/g oil.
  • the characteristics of the final oil according to this example are similar to the characteristics set forth in Table 26.
  • ingredients contained in the final oil include 3000 ppm Rosemary Extract; 1700 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 250 ppm ascorbyl palmitate.
  • the potency was obtained via the following protocol AOCS Ce 1b-89 (modified).
  • the marine/fishy aroma (smell) and marine/fishy aromatics (taste) sensory values were determined according to the method as described in Sensory Evaluation Techniques, Meilgaard et al., CRC Press; 4 edition (Dec. 13, 2006).
  • a panel of 8-18 experienced people tasted and/or smelled a sample of the final oils according to examples 10a-e and 11a-b. Each of these people determined the value of the sample. Afterwards all the values were averaged arithmetically and the result were rounded up or down to the next number.
  • a value of ⁇ 1.5 fishy/marine aroma and ⁇ 2.5 fishy/marine aromatics is expected to be perceivable by the general population.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be blended with HOSO to achieve a final oil with DHA content of at least about 400 mg/g oil.
  • Typical characteristics of the final oil according to this example are set forth in Table 28.
  • the 2400 ppm mixed tocopherol came from 2000 ppm of added tocopherols (includes 900 ppm that was added upstream), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70, and from the mixed tocopherols contained in the 4000 ppm of TAP 1010 Sun that was added to the oil.
  • TAP1010 Sun provided 400 ppm ascorbyl palmitate, 400 ppm mixed tocopherols, and 1600 ppm sunflower lecithin.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be further blended with HOSO to achieve final oil with DHA content of at least about 400 mg/g oil.
  • the characteristics of final oil according to this example are similar to the characteristics set forth in Table 28.
  • ingredients contained in the final oil include 2000 ppm Rosemary Extract; 1700 ppm mixed tocopherols (includes 900 ppm that was added upstream), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 250 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 3000 ppm Rosemary Extract; 1700 ppm mixed tocopherols (includes 900 ppm that was added upstream), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 250 ppm ascorbyl palmitate.
  • ingredients contained in the final oil include 5000 ppm Rosemary Extract; 1700 ppm mixed tocopherols (includes 900 ppm that was added upstream), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70; and 250 ppm ascorbyl palmitate.
  • a crude oil can be obtained from Schizochytrium sp via the processes described in, for example, WO 91/007498, WO 94/08467, WO 03/105606, and WO2011/153246.
  • the crude oil can be further processed via refining, bleaching, and deodorizing to obtain refined oils.
  • the refined oil can be further blended with organic HOSO, which is available from, for example, Adams Vegetables Oils, Inc, (Arbuckle, Calif.) to achieve a final oil with DHA content of at least about 400 mg/g oil.
  • Typical characteristics of the final oil according to this example are set forth in Table 29.
  • ppm Rosemary Extract which is available from, for example, Vitiva (Markovic, Slovenia) under the trade name Inolens® 4 organic and Naturex, (Avumble, France) under the trade name organic StabilEnhance® OSR-4; and 1700 ppm mixed tocopherols (includes 900 ppm that was added during upstream processing), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • ingredients contained in the final oil include 2000 ppm Rosemary Extract, which is available from, for example, Vitiva (Markovic, Slovenia) under the trade name Inolens® 4 organic and Naturex, (Avumble, France) under the trade name organic StabilEnhance® OSR-4; and 2400 ppm mixed tocopherols (includes 900 ppm that was added upstream), which is available from, for example, Vitablend (Wolvega, the Netherlands) under the trade name TocoblendTM L70.
  • the potency was obtained via the following protocol AOCS Ce 1b-89 (modified).
  • the marine/fishy aroma (smell) and marine/fishy aromatics (taste) sensory values were determined according to the method as described in Sensory Evaluation Techniques, Meilgaard et al., CRC Press; 4 edition (Dec. 13, 2006).
  • a panel of 8-18 experienced people tasted and/or smelled a sample of the oils according to examples 13, 14, and 15. Each of these people determined the value of the sample. Afterwards all the values were averaged arithmetically and the result was rounded up or down to the next number.
  • a value of ⁇ 1.5 fishy/marine aroma and ⁇ 2.5 fishy/marine aromatics is expected to be perceivable by the general population.

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