Edible oils with reduced content of mineral oil saturation hydrocarbons (MOSH) and methods for their production

Abstract

The present disclosure describes nutritional oils and methods of producing nutritional oils including a LC-PUFA and having a MOSH content of less than or equal to 7 ppm.

Description

NUTRITIONAL OILS HAVING REDUCED MOSH CONTENT AND METHODS OF MAKING THE SAMETECHNICAL FIELD

[0001] The present disclosure relates to microbial oils and nutritional oil compositions having reduced levels of mineral oil saturated hydrocarbons (MOSH). The present disclosure also relates to methods of making microbial oils and nutritional oil compositions having reduced levels of MOSH.BACKGROUND

[0002] Mineral oil includes a complex mixture of hydrocarbons having a variety of structures and chain lengths. Mineral oil hydrocarbons (MOH) include mineral oil saturated hydrocarbons (MOSH) and mineral oil aromatic hydrocarbons (MOAH). MOSH can include paraffins (openchain, saturated, optionally branched hydrocarbons) and naphthenes (cyclic, saturated, optionally highly alkylated, hydrocarbons), for example. MOAH can include mono- or polycyclic aromatic, optionally highly alkylated, hydrocarbons, for example.

[0003] MOH is a known food contaminant that is of particular concern in infant nutrition products. While MOAH is considered to be more toxic than MOSH, both are the subject of continued evaluation to determine their long-term effects on human health. MOH food contamination can originate from a variety of sources, such as packaging materials (e.g., plastics, adhesives, paper products, inks, etc.), manufacturing equipment (e.g., lubricants, cleaning agents, etc.), environmental contamination, etc.

[0004] Docosahexaenoic acid (DHA) and arachidonic acid (ARA) are long-chain polyunsaturated fatty acids (LC-PUFAs), defined as polyunsaturated fatty acids (PUFAs) having 20 to 22 carbons, that can influence infant growth and development. DHA is the most abundant omega-3 fatty acid in the brain and the retina. ARA is the primary omega-6 fatty acid in the brain. It is believed that DHA and ARA have positive effects on a child’s development when provided together and in efficacious amounts, during infancy. DHA is believed to support brain and cognition development, mental adaptability and problem solving, visual development, and attention and information processing. ARA is believed to play a role in supporting development of a healthy immune system, bone formation, and blood flow and blood vessel development.

[0005] As both DHA and ARA are believed to positively influence infant growth and development, these nutritional oils are often included in infant nutrition products. However, DHA and ARA products are also susceptible to MOSH and MOAH contamination from packaging,manufacturing equipment, etc. As such, there is a desire to minimize contamination of beneficial nutritional oils (e.g., LC-PUFA-containing oils) with potentially harmful MOH, including both MOSH and MOAH.BRIEF SUMMARY

[0006] The present disclosure describes nutritional oils including arachidonic acid (ARA) in an amount greater than or equal to 300 mg / g based on a total weight of the nutritional oil, free fatty acids in an amount greater than or equal to 0.1 wt% based on the total weight of the nutritional oil, and less than 7 ppm MOSH based on the total weight of the nutritional oil.

[0007] The present disclosure further describes nutritional oils including ARA in an amount greater than or equal to 300 mg / g based on the total weight of the nutritional oil, less than 0.1 wt% free fatty acids based on the total weight of the nutritional oil, and less than or equal to 3 ppm MOSH based on the total weight of the nutritional oil.

[0008] The present disclosure further describes nutritional oil blends including a first nutritional oil including ARA blended with a second nutritional oil including DHA, where the nutritional oil blend includes ARA and DHA at a weight ratio of from 0.5 to 3 ARA / DHA, and where the nutritional oil blend includes MOSH in an amount less than or equal to 4 ppm.

[0009] The present disclosure also describes methods of producing a nutritional oil including combining a LC-PUFA-containing oil with an organic solvent, removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude nutritional oil, and recovering at least a portion of the organic solvent removed from the LC-PUFA-containing oil using a recovery solvent, where the recovery solvent includes less than 50 wt% of mineral oil based on a total weight of the recovery solvent.DETAILED DESCRIPTION

[0010] Although the following detailed description contains many specifics for the purpose of illustration, a person of ordinary skill in the art will appreciate that many variations and alterations to the following details can be made and are considered to be included herein. Accordingly, the following embodiments are set forth without any loss of generality to, and without imposing limitations upon, any claims set forth. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0011] As used in this written description, the singular forms “a,” “an,” and “the” include express support for plural referents unless the context clearly dictates otherwise. Thus, for example, reference to “a fatty acid” or “the fatty acid” can include a plurality of such fatty acids.

[0012] Unless otherwise specified, all numerical parameters are to be understood as being prefaced and modified in all instances by the term “about,” in which the numerical parameters possess the inherent variability characteristic of the underlying measurement techniques used to determine the numerical value of the parameter.

[0013] Concentrations, amounts, and other numerical data may be expressed or presented herein in a range format. It is to be understood that such a range format is used merely for convenience and brevity and thus should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. As an illustration, a numerical range of “1 to 5” should be interpreted to include not only the explicitly recited values of 1 to 5, but also include individual values and subranges within the indicated range. Thus, included in this numerical range are individual values such as 1 , 2, 3, 4, and 5 and sub-ranges such as from 1 to 3, from 2 to 4, from 3 to 5, etc. This same principle applies to ranges reciting only one numerical value as a minimum or a maximum. Furthermore, such an interpretation should apply regardless of the breadth of the range or the characteristics being described.

[0014] Reference throughout this written description to “an example” means that a particular feature, component, or characteristic described in connection with the example is included in at least one embodiment. Thus, appearances of the phrases “in an example” or “in one example” in various places throughout this written description are not necessarily all referring to the same embodiment.Oil Sources

[0015] The methods described herein can be performed, and the microbial oils or nutritional oil compositions described herein can be obtained, via a suitable biomanufacturing process employing one or more suitable plants or microorganisms. For example, microbial oils or nutritional oils can be produced and obtained from plants, plant seeds, microalgae, fungi (including yeast), bacteria, and / or protists.

[0016] In some specific examples, the nutritional oils can be extracted or otherwise obtained from a plant genetically modified to produce, or increase production of at least one LC-PUFA, such as a LC-PUFA-containing coconut oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil,rapeseed oil (canola oil), safflower oil, sesame oil, soybean oil, sunflower oil, camelina oil, etc. Additional examples and description can be found in U.S. Patent Publication Nos. 2015 / 0152450, 2013 / 0101723, 2013 / 0150599, 2010 / 0313309, 2008 / 0050505, and 2007 / 0220634, each of which is incorporated herein by reference. Plants can be grown and the oil extracted from the plants, or seeds thereof, using methods known in the art.

[0017] In some specific examples, the microbial oils described herein can be produced, or largely produced, by a fungus. In some additional specific examples, the microbial oils described herein can be produced, or largely produced, by a fungus of the order Mortierellales.

[0018] In some specific examples, the microbial oils described herein can be produced, or largely produced, by a fungus of the genus Mortierella. In some examples, the microbial oils described herein can be produced, or largely produced by a fungus comprising Mortierella alpina, Mortierella vinacea, Mortierella polycephala, Mortierella elongata, or a combination thereof.

[0019] In some specific examples, the microbial oils described herein can be produced, or largely produced, by a microalgae. In some examples, the microalgae can be or include a dinoflagellate, such as Crypthecodinium cohnii, for example.

[0020] In some additional specific examples, the microbial oils described herein can be produced, or largely produced, by a Thraustochytrid, which, at the time of this disclosure, is classified as a type of microalgae.

[0021] For purposes of the present disclosure, strains described as Thraustochytrids include organisms of the taxonomic order Thraustochytrialies. In some further examples, Thraustochytrids can include organisms of the taxonomic family Thraustochytriidae (or Thraustochytriaceae). In still further examples, Thraustochytrids can include organisms of the taxonomic genus Thraustochytrium, Ulkenia, Schizochytrium, Japonochytrium, Aplanochytrium, Althornia, Elina, Aurantiochytrium, Oblongichytrium, Botryochytrium, Parietichytrium, Sicyoidochytrium, or a combination thereof.

[0022] In some further examples, the microbial oils described herein can be produced, or at least largely produced, from a microorganism of a species, or a microorganism derived from a species, of the genus Thraustochytrium, Ulkenia, Schizochytrium, Japonochytrium, and / or Aplanochytrium.

[0023] Additional species of microorganisms that can be used to produce microbial oils can be found in U.S. Patent No. 8486267, which is incorporated herein by reference.Culture and Fermentation

[0024] The microorganisms described herein can be cultured in large scale industrial bioreactors. The microorganisms can be cultured under conditions that increase biomass and / or production of a compound of interest (e.g., total lipid content, total LC-PUFA content, etc.). For example, the production of desirable lipids can be enhanced by culturing cells according to methods that involve a shift of one or more culture conditions to obtain higher quantities of desirable polyunsaturated fatty acids (PUFAs), or in particular, long chain PUFAs (LC-PUFAs) having carbon chain lengths of 20 to 22 carbon atoms.

[0025] In further detail, LC-PUFAs can typically be produced by culturing the microorganisms in a two-phase process that includes a growth phase, in which cell biomass is increased, followed by a production phase, in which the biomass synthesizes lipids. The lipid component can be considered a microbial oil and can be separated from the biomass for use as a nutritional oil or for use as a component of a nutritional oil composition. The particular growth media and culture conditions employed will depend on the particular microorganism being cultivated.

[0026] Generally, the microorganisms can typically be cultivated to achieve a biomass concentration of at least 20 g / L by the end of the growth phase. In some further examples, the microorganisms can be cultivated to achieve a biomass concentration of at least 40 g / L, at least 60 g / L, at least 80 g / L, at least 90 g / L, or at least 100 g / L by the end of the growth phase. Biomass concentration can be determined based on the mass of a washed (e.g., to remove residual salts, carbohydrates, etc.) and lyophilized biomass obtained per unit volume of fermentation media.

[0027] The microorganisms can typically be cultivated to achieve a lipid content (or fat content or oil content) by the end of the production phase of at least 20 wt.% based on a total dry cell weight of the microorganism biomass. In some further examples, the microorganisms can be cultivated to achieve a lipid content (or fat content or oil content) by the end of the production phase of at least 30 wt.%, at least 40 wt.%, at least 50 wt.%, at least 55 wt.%, at least 60 wt.%, or at least 65 wt.% based on the total dry cell weight of the biomass. Dry cell weight is based on the mass of a washed (e.g., to remove residual salts, carbohydrates, etc.) and lyophilized biomass.

[0028] In some examples, the volume of the culture media can be at least 2 liters, at least 10 liters, at least 50 liters, at least 100 liters, at least 200 liters, at least 200 liters, at least 1000 liters, at least 10,000 liters, at least 20,000 liters, at least 50,000 liters, at least 100,000 liters, at least 150,000 liters, at least 200,000 liters, or at least 250,000 liters. In some examples, theculture media is from 2 liters to 300,000 liters, from 10 liters to 200,000 liters or 250,000 liters, from 50 liters to 100,000 liters or 150,000 liters, from 100 liters to 50,000 liters, from 500 liters to 20,000 liters, or from 1000 liters to 10,000 liters.

[0029] The microorganism cells can be cultivated for a cultivation period anywhere from 1 day to 60 days. In some examples, the microorganism cells can be cultivated for a cultivation period of from 1 day to 14 days, from 2 days to 13 days, from 3 days to 12 days, from 4 days to 11 days, from 5 days to 10 days, from 6 days to 9 days, or from 7 days to 8 days.

[0030] In some examples, cultivation can include aeration-shaking culture, shaking culture, stationary culture, batch culture, semi-continuous culture, continuous culture, rolling batch culture, wave culture, or the like. In some additional examples, cultivation can be performed used a conventional agitation-fermenter, a bubble column fermenter (batch or continuous), a wave fermenter, or the like. Cultures can be aerated using a variety of methods, such as shaking, mixing, bubbling, etc.

[0031] Additional methods and details related to culture and fermentation can be found in U.S. Patent Nos. 8,669,090; 9,045,785; 9,453,172; and 10,435,725 and U.S. Patent Publication No. 2021 / 0171991 , each of which is incorporated by reference.Pasteurization

[0032] Optionally, the biomass resulting from the formation process can be pasteurized to kill the cells and inactivate undesirable substances present in the biomass. For example, the biomass can be pasteurized to inactivate substances that can lead to the degradation of desirable lipids or other compounds. The biomass can be pasteurized in the fermentation media or isolated from the fermentation media for pasteurization. Pasteurization can be performed by heating the biomass and / or fermentation media to temperature sufficient to kill the cells and / or inactivate certain undesirable substances.

[0033] In some examples, pasteurization can be performed as described in U.S. Patent Publication No. 2005 / 0220958, which is incorporated herein by reference.

[0034] In some additional examples, pasteurization can be performed at a temperature of from 50°C to 90°C for a pasteurization period. In other examples, pasteurization can be performed at a temperature of from 50°C to 70°C, from 60°C to 80°C, or from 70°C to 90°C for a pasteurization period.

[0035] The pasteurization period can typically be less than or equal to 120 minutes. In some examples, the pasteurization period can be from 30 minutes to 120 minutes. In some specific examples, the pasteurization period can be from 30 minutes to 90 minutes, from 45 minutes to105 minutes, or from 60 minutes to 120 minutes. In still additional examples, the pasteurization period can be from 45 minutes to 75 minutes, or from 55 minutes to 90 minutes.

[0036] Pasteurization can be performed using a variety of suitable methods, including indirect and / or direct heating methods. In some examples pasteurization can be performed using an indirect heating method, such as via a heat transfer jacket or plate, for example. In some examples, pasteurization can be performed using a direct heating method, such as direct steam injection, for example.Harvesting

[0037] Optionally, the biomass can be harvested according to a variety of conventional methods. As non-limiting examples, the biomass can be collected using centrifugation (e.g., solid-ejecting centrifugation), filtration (e.g., cross-flow filtration), and / or other suitable harvesting methods. In some examples, harvesting can also employ the use of a precipitation agent (e.g., sodium phosphate or calcium chloride) to accelerate collection of the biomass.

[0038] In some examples, the biomass can also be washed with water. In some examples, the biomass can be concentrated to achieve a desired solids content. Additional methods and details related to harvesting biomass are also discussed in U.S. Patent No. 5,130,242, U.S. Patent Publication No. 2002 / 0001833, and International Patent Publication No.W0 1997 / 037032, each of which is incorporated herein by reference.Hydrolysis

[0039] Optionally, cell hydrolysis (i.e. , biomass cell disruption) can be performed using a variety of methods. In some examples, cell hydrolysis can be performed using chemical, enzymatic, and / or mechanical methods.

[0040] Chemical hydrolysis can be performed in a variety of ways, such as via acid hydrolysis, base hydrolysis, detergent hydrolysis, or the like. In some examples, chemical hydrolysis can be performed by acid hydrolysis by adding an acid to the biomass mixture. Typically, acid hydrolysis can include washing the biomass with water. Acid can be added to the washed biomass mixture. In some examples, the biomass is not dried prior to adding the acid. In other examples, the biomass is dried prior to adding the acid. Non-limiting examples of acids that can be used in acid hydrolysis can include sulfuric acid, hydrochloric acid, phosphoric acid, hydrobromic acid, nitric acid, perchloric acid, or other strong acids.

[0041] The acid can be added to the biomass mixture in a sufficient amount and for a sufficient incubation period to hydrolyze the cells. In some examples, the acid can be added to thebiomass mixture in an amount to achieve a final concentration of acid of from 100 mM to 200 mM. In some additional examples, the acid can be added to the biomass mixture in an amount to achieve a final concentration of acid of from 100 mM to 140 mM, from 120 mM to 160 mM, from 140 mM to 180 mM, or from 160 mM to 200 mM.

[0042] The incubation period to hydrolyze the cells is further impacted by the temperature at which acid hydrolysis is conducted. In some examples, acid hydrolysis can be conducted at a temperature of from 30°C to 200°C. In some further examples, acid hydrolysis can be conducted at a temperature of from 30°C to 80°C, from 50°C to 100°C, from 75°C to 125°C, from 100°C to 150°C, from 125°C to 175°C, or from 150°C to 200°C.

[0043] The incubation period can be a period that is sufficient at the concentration of acid and the temperature to hydrolyze the cells. For example, incubating the mixture at a higher temperature can result in the hydrolysis proceeding at a faster rate (i.e. , requiring a shorter period of time for hydrolysis). Similarly, in some examples, incubating the mixture at a higher concentration of acid can result in the hydrolysis proceeding at a faster rate.

[0044] In some examples, hydrolysis of the cells in the biomass can be performed using enzymatic hydrolysis. For example, the biomass can be contacted with one or more enzymes under conditions that cause disruption of the cells. In some examples, the enzyme can be a protease. One non-limiting example of a protease that can be used for enzymatic hydrolysis of the biomass cells is ALCALASE 2.4L FG (Novozymes; Franklinton, N.C.). In some examples, the biomass can be washed prior to the enzymatic hydrolysis.

[0045] The microorganisms can typically be fermented to float in aqueous media. The fermentation media can be gravity settled in the fermenter and the media can be decanted or otherwise removed to provide the desired concentration of the biomass. Alternatively, the fermentation media can be concentrated by centrifugation to provide the desired concentration of the biomass. In some examples, the biomass can be contacted with one or more enzymes while the microorganisms are in the fermentation medium. Optimum temperature, time, pH and enzyme concentration depend on the specific enzyme employed. Enzymatic hydrolysis can be performed with or without the use of a surfactant.

[0046] Other methods of hydrolysis can also be employed. Non-limiting examples can include bead milling, sonication, rapid decompression, high-shear mechanical methods, homogenization, ultrasound, French press, cold-pressing, osmotic shock, heating, drying, pressure oscillation, solvent extraction, expression of an autolysis gene, the like, or a combination thereof. In some examples, hydrolysis of the biomass cells can be performed usinga combination of chemical, enzymatic, and / or mechanical methods, either sequentially or concurrently.

[0047] Other methods or details related to hydrolysis can be found in U.S. Patent No. 9,408,404 and U.S. Patent Publication Nos. 2022 / 0145211 and 2022 / 0154098, each of which is incorporated herein by reference.Extraction

[0048] Lipids can be extracted from a plant or microbial biomass in a variety of ways. As used herein, the term “lipid” includes phospholipids, free fatty acids, esters of fatty acids, acylglycerols (e.g., triacylglycerols, diacylglycerols, monoacylglycerols), lysophospholipids, soaps, phosphatides, sterols, sterol esters, carotenoids, xanthophylls, hydrocarbons, and the like. The lipid-rich material that is extracted from the biomass can generally be referred to as a LC-PUFA-containing oil, or as a crude oil.

[0049] Different types or components of the lipids can be extracted, depending on the extraction technique that is used. In some examples, the lipid can be isolated from a plant or microorganism using standard techniques, without being subjected to further refinement or purification. In some examples, the lipid can be isolated using physical and / or mechanical extraction methods, such as, but not limited to, extraction via homogenization or pressing.

[0050] In other examples, the lipid can be extracted using solvent extraction. Where solvent extraction is used, a variety of organic solvents can be employed. Non-limiting examples can include hexane, isopropyl alcohol, methylene chloride, dodecane, methanol, ethylated oil, supercritical carbon dioxide, the like, or a combination thereof. Polar lipids (e.g., phospholipids) are generally extracted with polar solvents (e.g., chloroform / methanol). Non-polar lipids (e.g., triacylglycerols) are generally extracted with non-polar solvents (e.g., hexane). In some specific examples, where solvent extraction is used, the extraction solvent can be or include hexane.

[0051] In solvent extraction, the organic solvent and biomass can be mixed for a period of time suitable to extract lipids from the biomass. For example, the organic solvent and biomass can be mixed for a period of time of from 10 minutes to 2 hours or more. Subsequently, the lipid can be separated from the remaining components of the mixture by centrifugation, filtration, the like, or a combination thereof.

[0052] In other examples, extraction can be performed using reduced amounts of organic solvent as compared to the amounts typically used to extract lipids from whole dry biomass. For example, the ratio of biomass to organic solvent needed to extract lipids from whole drybiomass is typically 1 :4 or greater. Thus, the reduced amount of organic solvent can provide a ratio of biomass to organic solvent of less than 1 :4.

[0053] Optionally, lipid extraction can be performed using an edible oil or biofuel. In some examples, the edible oil can be a plant oil, such as coconut oil, palm oil, canola oil, sunflower oil, soy oil, corn oil, olive oil, safflower oil, palm kernel oil, cottonseed oil, an alkylated derivative thereof, the like, or a combination thereof. In some additional examples, the edible oil can be a microbial oil, an alkylated derivative, the like, or combination thereof. As used herein, “biofuel” refers to any fuel, fuel additive, aromatic, and / or aliphatic compound derived from a biomass starting material. In some examples, biofuels can be derived from plant sources or microbial sources. Non-limiting examples of sources of biofuels can include microalgae, corn, switchgrass, sugarcane, sugarbeet, rapeseed, soybean, or the like. As used herein, the term organic solvent does not include biofuels as that term is defined herein and does not include an edible oil.

[0054] Optionally, where an edible oil and / or biofuel is used to extract lipids from the biomass, the edible oil and / or biofuel is not fully removed, or, in some cases, substantially unremoved, from the lipid extract. A subsequent fractionation of the extracted oil, wherein the added oil or biofuel stays with only one of the oil fractions, is not considered removal of the edible oil or biofuel from the lipid extract. In some examples, after recovery, the crude oil may be combined with other edible oils or biofuels for use as, or incorporated into, one of more or the nutritional oil compositions described herein. The edible oil or biofuel may be added to the long chain polyunsaturated fatty acid (LC-PUFA)-containing oil during the extraction step as an alternative to, or in addition to, combining with the LC-PUFA-containing oil after the recovery process. In some cases, adding the edible oil or biofuel during the extraction step can assist in the separation of the LC-PUFA-containing oil from the biomass.

[0055] In traditional methods that rely on organic solvent extraction to separate the LC-PUFA- containing oil from the biomass, the organic solvent is removed from the LC-PUFA-containing oil after recovery, although trace amounts of the organic solvent may be left behind. In the methods described herein, where an edible oil or biofuel is employed to assist with the LC- PUFA-containing oil extraction, optionally at least 80 wt.% of the edible oil or biofuel added during the extraction step can remain with the extracted LC-PUFA-containing oil and can be incorporated into the final nutritional oil composition. In other examples, at least 85 wt.%, at least 90 wt.%, at least 95 wt.%, at least 98 wt.%, or 100 wt.% of the edible oil or biofuel used in the extraction step can remain with the extracted LC-PUFA-containing oil and can be incorporated into the final nutritional oil composition.

[0056] Alternatively, or additionally, the LC-PUFA-containing oil can be extracted using mechanical methods. For example, the hydrolyzed biomass can be centrifuged to separate the LC-PUFA-containing oil from the other components. In some examples, the LC-PUFA- containing oil can be included in an upper layer of the centrifuged material and can be isolated by suction or decanting from the residual material.Refining

[0057] The nutritional oil or microbial oil isolated from the other components of the plant or microbial biomass mixture can be referred to as a crude nutritional oil, which can include the LC-PUFAs produced by the plants and / or microorganisms, as well as a variety of other components extracted with the LC-PUFAs, such as sterols, other fatty acids, residual edible oils / biofuels employed in the extraction process, etc. In some examples, the nutritional oil compositions described herein can be or include a crude nutritional oil extracted from the biomass.

[0058] Optionally, the crude nutritional oil can be subjected to further refining processes to produce a refined oil or a final oil. In some examples, the nutritional oil compositions described herein can be or include a refined nutritional oil or a final nutritional oil. In some specific examples, the refined nutritional oil or final nutritional oil can be a refined microbial oil or a final microbial oil.

[0059] Refining of the crude nutritional oil can include any of a variety of standard vegetable oil refining steps, such as degumming, neutralization / caustic refining, bleaching, deodorization, winterization / dewaxing, etc. When performed, refining can remove a variety of impurities or minor components, such as, but not limited to, free fatty acids, water, impurities, colorants, phospholipids, minerals, carotenoids, sterols, tocopherols / tocotrienols, waxes, residual cell debris, residual solvent, etc.

[0060] In further detail, degumming can be performed to remove phospholipids / gums from the crude oil, as desired. Degumming can also help to minimize refining losses and to minimize decomposition of the oil, for example. In some examples, degumming can include water degumming and / or acid degumming.

[0061] Neutralization is typically performed after degumming to remove free fatty acids. Neutralization typically includes saponifying the free fatty acids using an alkaline solution and extracting the resulting soap in a water phase, which can be easily separated from the oil. Neutralization can also be used to remove phospholipids, oxidized components in the oil, metal ions, colorants (e.g., color pigments), insoluble impurities, etc.

[0062] Bleaching is typically performed after neutralization and can be used to remove colorants (e.g., color pigments) from the oil. Bleaching can be performed by adding bleaching clays or other suitable adsorbents to adsorb colorants present in the oil. The bleaching clay or other adsorbent can then be removed by filtration.

[0063] Winterization or dewaxing is typically performed after bleaching and can be employed to removed waxes or saturated fatty acids from the oil. The presence of saturated fatty acids can cause the oil to have a cloudy or hazy appearance. Typically, winterization can be performed by chilling the oil to precipitate saturated fatty acids from the oil. The precipitated solids can then be filtered and removed from the oil.

[0064] Deodorization is typically performed last in the overall oil refining process to ensure a clean feedstock. This process typically includes steam stripping the oil at high temperature under vacuum to help remove unwanted volatile compounds, off-odors, hydroperoxides, and other contaminants in the oil.

[0065] Other methods and details related to refining of the crude oils can be found in U.S. Patent Nos. 7,419,596 and 11 ,672,258, each of which is incorporated herein by reference.Minimizing Mineral Oil Hydrocarbons

[0066] Mineral oil hydrocarbons (MOH) can be introduced into the nutritional oil compositions in a variety of ways. As previously discussed, MOH can be introduced into the nutritional oil compositions via packaging materials, manufacturing equipment, environmental sources, etc. Accordingly, adjusting any one of these factors to minimize MOH contamination can be considered.

[0067] In some specific examples, minimizing MOH can be accomplished by addressing sources of MOH contamination in the manufacturing process. However, MOH can be an effective component in various aspects of the manufacturing process. For example, a mineral oil-based system is commonly employed to absorb and recover evaporated extraction solvent for subsequent reuse.

[0068] Specifically, the LC-PUFA-containing oil can be extracted from the plant or microbial biomass using an organic solvent. The organic solvent can be removed from the LC-PUFA- containing oil, typically by evaporation, and a solvent recovery system can be used to capture and reuse the organic solvent. In further detail, the effluent air leaving the solvent recovery system contains solvent vapor and can be introduced into a mineral oil-based absorber system to absorb the residual solvent from the effluent air before it is vented to the atmosphere. The effluent air can be directed into mineral oil-based system to rise through a solid packing materialto maximize the oil / vapor contact surface area. Mineral oil is introduced into the mineral oilbased system in a manner that it flows counter-currently to the effluent gas. Accordingly, the organic solvent carried by the effluent air can be absorbed by the mineral oil.

[0069] After absorbing the organic solvent, the mineral oil can be heated with steam under vacuum to strip out and condense the organic solvent for later reuse. The stripped mineral oil can then be cooled and circulated back to the mineral oil-based system for reuse. However, traces of mineral oil saturated hydrocarbons (MOSH) and / or mineral oil aromatic hydrocarbons (MOAH) can contaminate the organic extraction solvent, such as through aerosolization, for example. These trace amounts of MOSH and MOAH can then be transferred to the LC-PUFA- containing oil during the extraction process, rather than remaining with the extraction solvent, resulting in unwanted oil contamination. The amount of MOSH and / or MOAH contamination can depend on a variety of factors, such as run rate, vapor pressure, temperature, cleanliness of the packing phase, condition of the extraction solvent, condition of the mineral oil, etc.

[0070] Accordingly, the present disclosure describes a method of minimizing MOSH and / or MOAH in a nutritional oil by employing an alternative, but effective, recovery solvent. More specifically, the method can include replacing at least of portion of the mineral oil in the mineraloil based system. Thus, a recovery solvent including less than 50 wt% mineral oil based on the total weight of the recovery solvent can be used to recover used organic solvent and minimize MOH contamination of the nutritional oil. In further detail, the method can include combining a LC-PUFA-containing oil with an organic solvent, removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude nutritional oil, and recovering at least a portion of the organic solvent using a recovery solvent having less than 50 wt% mineral oil based on a total weight of the recovery solvent.

[0071] The LC-PUFA-containing oil can be any LC-PUFA-containing oil that is combined with an organic solvent (e.g., for extraction). In some examples, the LC-PUFA-containing oil can include eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (ARA), or a combination thereof. In some specific examples, the LC-PUFA-containing oil can include EPA. In some specific examples, the LC-PUFA-containing oil can include DHA. In some specific examples, the LC-PUFA-containing oil can include ARA.

[0072] A variety of organic solvents can be combined with the LC-PUFA-containing oil. Nonlimiting examples can include hexane, isopropyl alcohol, methylene chloride, dodecane, methanol, ethanol, acetone, ethylated oil, supercritical carbon dioxide, the like, or a combination thereof. In some specific examples, the organic solvent can include hexane. In some additionalspecific examples, the organic solvent can include isopropyl alcohol. In some further specific examples, the organic solvent can include ethanol and / or methanol.

[0073] In some examples, the organic solvent can include at least a portion of a recovered organic solvent that was previously combined with a LC-PUFA-containing oil and subsequently recovered using a solvent recovery process. In some examples, the organic solvent can include greater than 40 wt%, greater than 50 wt%, greater than 60 wt%, or greater than 70 wt% recovered organic solvent.

[0074] The recovered organic solvent can typically include less than 100 ppm MOSH. In some additional examples, the recovered organic solvent can include less than 50 ppm MOSH, less than 40 ppm MOSH, less than 30 ppm MOSH, less than 20 ppm MOSH, less than 10 ppm MOSH, less than 5 ppm MOSH, or less than 1 ppm MOSH.

[0075] Additionally, the recovered organic solvent can typically include less than 100 ppm MOAH. In some additional examples, the recovered organic solvent can include less than 50 ppm MOAH, less than 40 ppm MOAH, less than 30 ppm MOAH, less than 20 ppm MOAH, less than 10 ppm MOAH, less than 5 ppm MOAH, or less than 1 ppm MOAH.

[0076] MOSH and MOAH can be detected and measured using any suitable analytical methodology. As a non-limiting example, MOSH and MOAH can be analyzed using gas chromatography (GO) with flame ionization detection (FID).

[0077] Combining the organic solvent and / or recovered organic solvent with the LC-PUFA- containing oil can typically be performed to extract the LC-PUFA-containing oil from a biomass, such as a plant biomass or a microbial biomass, for example. In specific examples, the organic solvent and / or recovered organic solvent can be combined with the LC-PUFA-containing oil to extract the LC-PUFA-containing oil from a plant biomass. In some additional examples, the organic solvent and / or recovered organic solvent can be combined with the LC-PUFA- containing oil to extract the LC-PUFA-containing oil from a microbial biomass.

[0078] The organic solvent and / or recovered organic solvent can be removed from the LC- PUFA-containing oil to obtain a crude nutritional oil. The crude nutritional oil can typically include less than 5 wt% organic solvent after removal. In some further examples, the crude nutritional oil can include less than 3 wt%, less than 1 wt%, less than 0.5 wt%, less than 0.1 wt%, less than 0.05 wt%, less than 0.01 wt%, less than 0.005 wt%, or less than 0.001 wt% organic solvent. Thus, the organic solvent can at least partially be removed from the LC-PUFA- containing oil. In some further examples, the organic solvent can be substantially removed from the LC-PUFA-containing oil.

[0079] To prevent the organic solvent from being evacuated to the atmosphere, a solvent recovery system can be used to recapture and reuse the organic solvent. Furthermore, the solvent recovery system typically employs a recovery solvent to absorb the organic solvent. Typically, the recovery solvent includes less than 50 wt% mineral oil based on the total weight of the recovery solvent. In some additional examples, the recovery solvent includes less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, or less than 1 wt% mineral oil based on the total weight of the recovery solvent. This can minimize the contribution of the recovery solvent to the MOSH and / or MOAH levels of the crude nutritional oil.

[0080] In additional examples, the recovery solvent can include an edible oil, such as coconut oil, palm oil, canola oil, sunflower oil, soy oil, corn oil, olive oil, safflower oil, palm kernel oil, cottonseed oil, medium chain triglycerides, an alkylated derivative thereof, the like, or a combination thereof. In further examples, the recovery solvent can include a biofuel. In some specific examples, the recovery solvent can include medium chain triglycerides. In some specific examples, the recovery solvent can include canola oil. In some specific examples, the recovery solvent can include sunflower oil.

[0081] The organic solvent can be removed from the LC-PUFA-containing oil to provide a crude nutritional oil. In some examples, the nutritional oil can be the crude nutritional oil. In some further examples, the crude nutritional oil can be further processed to obtain a refined nutritional oil or final nutritional oil.

[0082] In some additional examples, minimizing MOSH and / or MOAH in a nutritional oil can be accomplished, or further accomplished, by replacing at least a portion of mineral oil used elsewhere in the manufacturing process (e.g., as an equipment lubricant) with a replacement equipment material, which can be the same as or different from the recovery solvent. Where employed, the equipment material typically includes less than 50 wt% mineral oil based on the total weight of the equipment material. In some additional examples, the equipment material includes less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, less than 5 wt%, or less than 1 wt% mineral oil based on the total weight of the equipment material. In some specific examples, the equipment material is an equipment lubricant.In additional examples, the equipment material can be the same material as the recovery solvent. In some alternative examples, the equipment material can be a different material from the recovery solvent. In some examples, the equipment material can include an edible oil, such as coconut oil, palm oil, canola oil, sunflower oil, soy oil, corn oil, olive oil, safflower oil, palm kernel oil, cottonseed oil, medium chain triglycerides, an alkylated derivative thereof, the like, or a combination thereof. In further examples, the equipment material can include a biofuel. Insome specific examples, the equipment material can include medium chain triglycerides. In some specific examples, the equipment material can include canola oil. In some specific examples, the recovery solvent can include sunflower oil.Compositions

[0083] The present disclosure describes a variety of nutritional oil compositions. In some examples, the nutritional oils can be a crude nutritional oil. In some examples, the nutritional oil can be a refined nutritional oil or final nutritional oil.

[0084] In some examples, the nutritional oil can be a crude nutritional oil and can include greater than 0.1 wt% free fatty acids based on a total weight of the nutritional oil. In still further examples, the crude nutritional oil can include greater than 0.15 wt%, greater than 0.2 wt%, greater than 0.25 wt%, greater than 0.3 wt%, greater than 0.35 wt%, greater than 0.4 wt%, greater than 0.45 wt%, greater than 0.5 wt%, or greater than 0.55 wt% free fatty acids based the total weight of the nutritional oil. In some additional examples, the nutritional oil can include free fatty acids in an amount of less than 0.5 wt%, less than 0.45 wt%, less than 0.4 wt%, less than 0.35 wt%, less than 0.3 wt%, less than 0.25 wt%, less than 0.2 wt%, less than 0.15 wt%, less than 0.1 wt%, or less than 0.05 wt% based on the total weight of the nutritional oil. In some specific examples, the nutritional oil can be a refined nutritional oil or a final nutritional oil and can include less than 0.1 wt% free fatty acids based on a total weight of the nutritional oil.

[0085] Free fatty acid content can be determined using any suitable analytical methodology. In some non-limiting examples, free fatty acid content can be determined using AOCS Ca 5a-40, or similar.

[0086] The nutritional oils described herein can typically have less than or equal to 7 ppm MOSH. In some additional examples, the nutritional oils described herein can include less than or equal to 5 ppm MOSH, or less than or equal to 3 ppm MOSH. In some specific examples, the nutritional oil compositions described herein can include MOSH in an amount less than or equal to 7 ppm, less than or equal to 6 ppm, less than or equal to 5 ppm, less than or equal to 4 ppm, less than or equal to 3 ppm, less than or equal to 2.5 ppm, less than or equal to 2 ppm, less than or equal to 1.5 ppm, less than or equal to 1 ppm, or less than or equal to 0.5 ppm based on the total weight of the nutritional oil. In some additional examples, the nutritional oil can include MOSH in an amount from 0.1 ppm to 7 ppm based on the total weight of the nutritional oil. In some further examples, the nutritional oil can include MOSH in an amount from 0.1 ppm to 2 ppm, from 0.5 ppm to 3 ppm, 1 ppm to 4 ppm, or 1.5 ppm to 5 ppm based on the total weight of the nutritional oil.

[0087] In some specific examples, the nutritional oils described herein can include MOAH in an amount less than or equal to 5 ppm, less than or equal to 4 ppm, less than or equal to 3 ppm, less than or equal to 2 ppm, less than or equal to 1.5 ppm, less than or equal to 1 ppm, or less than or equal to 0.5 ppm based on the total weight of the nutritional oil. In some additional examples, the nutritional oils described herein can include MOAH in an amount from 0.001 ppm to 2 ppm or 0.01 to 5 ppm based on the total weight of the nutritional oil.

[0088] MOSH and MOAH can be detected and measured using any suitable analytical methodology. As a non-limiting example, MOSH and MOAH can be isolated using solid-phase extraction technology and subsequently analyzed using gas chromatography with flame ionization detection. MOSH and MOAH can also be analyzed using DIN EN 16995: 2017-08, or similar.

[0089] The nutritional oils described herein can include a variety of long chain polyunsaturated fatty acids (LC-PUFAs). In some examples, the nutritional oils described herein can include arachidonic acid (ARA), docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), or a combination thereof. In some examples, the nutritional oils described herein can include ARA. In some examples, the nutritional oils described herein can include DHA. In some examples, the nutritional oils described herein can include EPA. In some further examples, the nutritional oils described herein can include ARA, DHA, and EPA.

[0090] It is noted that the nutritional oils typically include a large amount of acylglycerols, such as triglycerides, diglycerides, and monoglycerides. In some examples, the nutritional oil can include triglycerides in an amount greater than or equal to 50 wt%, greater than or equal to 55 wt%, greater than or equal to 60 wt%, greater than or equal to 65 wt%, greater than or equal to 70 wt%, greater than or equal to 75 wt%, greater than or equal to 80 wt%, greater than or equal to 85 wt%, greater than or equal to 90 wt%, or greater than or equal to 95 wt% based on the total weight of the nutritional oil.

[0091] The amount of triglyceride in the nutritional oil can be measured using any suitable analytical methodology. In one non-limiting example, the triglyceride content can be measured using1H-NMR.

[0092] In further detail, fatty acids, such as PUFAs, can be bound to a glycerol backbone at one of three positions to form an acylglycerol. The two terminal hydroxyls along the glycerol backbone are referred to as the sn-1 and sn-3 positions. The central hydroxyl is referred to as the sn-2 position. Typically, the PUFAs are bound to a glycerol backbone to form an acylglycerol. Thus, when referring to any particular PUFA, it is to be understood that such 1reference includes both PUFAs bound to a glycerol backbone forming an acylglycerol and free PUFAs, unless otherwise indicated.

[0093] In some examples, the nutritional oils described herein can include a LC-PUFA in an amount greater than or equal to 300 mg / g based on the total weight of the nutritional oil. In some additional examples, the nutritional oil can include a LC-PUFA in an amount greater than or equal to 350 mg / g, greater than or equal to 400 mg / g, greater than or equal to 450 mg / g, greater than or equal to 500 mg / g, or greater than or equal to 550 mg / g based on the total weight of the nutritional oil. In some further examples, the LC-PUFA can be present in the nutritional oils in an amount from 300 mg / g to 400 mg / g, from 350 mg / g to 450 mg / g, from 400 mg / g to 500 mg / g, from 450 mg / g to 550 mg / g, from 500 mg / g to 600 mg / g, from 550 mg / g to 650 mg / g based on the total weight of the nutritional oil.

[0094] The amount of LC-PUFA or other fatty acid in the nutritional oils can be determined using any suitable analytical methodology. In some non-limiting examples, the fatty acid (e.g., LC-PUFA) can be quantified against reference standard material using AOCS Ce 1 b-89, or similar.

[0095] In some specific examples, the nutritional oils can include ARA. In some further examples, the nutritional oils can include ARA in an amount greater than or equal to 300 mg / g based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include ARA in an amount greater than or equal to 350 mg / g, greater than or equal to 400 mg / g, greater than or equal to 450 mg / g, greater than or equal to 500 mg / g, or greater than or equal to 550 mg / g based on the total weight of the nutritional oil. In still additional examples, the nutritional oils can include ARA in an amount from 300 mg / g to 400 mg / g, from 350 mg / g to 450 mg / g, from 400 mg / g to 500 mg / g, from 450 mg / g to 550 mg / g, or from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0096] In some specific examples, the nutritional oils can include dihomo-gamma-linolenic acid (DGLA) (20:3 n-6). In some further examples, the nutritional oils can include DGLA in an amount greater than or equal to 10 mg / g based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include DGLA in an amount greater than or equal to 15 mg / g, greater than or equal to 20 mg / g, greater than or equal to 25 mg / g, or greater than or equal to 30 mg / g based on the total weight of the nutritional oil. In still additional examples, the nutritional oils can include DGLA in an amount from 10 mg / g to 20 mg / g, from 15 mg / g to 25 mg / g, from 20 mg / g to 30 mg / g, or from 25 mg / g to 35 mg / g based on the total weight of the nutritional oil.

[0097] In some specific examples, the nutritional oils can include gamma-linolenic acid (GLA) (18:3 n-6). In some further examples, the nutritional oils can include GLA in an amount greater than or equal to 10 mg / g based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include GLA in an amount greater than or equal to 15 mg / g, greater than or equal to 20 mg / g, greater than or equal to 25 mg / g, greater than or equal to 30 mg / g, or greater than or equal to 35 mg / g based on the total weight of the nutritional oil. In still additional examples, the nutritional oils can include GLA in an amount from 10 mg / g to 20 mg / g, from 15 mg / g to 25 mg / g, from 20 mg / g to 30 mg / g, from 25 mg / g to 35 mg / g, or from 30 mg / g to 40 mg / g based on the total weight of the nutritional oil.

[0098] In some examples, the nutritional oil can include at least 30 mg / g palmitic acid based on the total weight of the nutritional oil. In still additional examples, the nutritional oil can include at least 40 mg / g, at least 50 mg / g, at least 60 mg / g, at least 70 mg / g, at least 80 mg / g, or at least 90 mg / g palmitic acid based on the total weight of the nutritional oil. In some further examples, the nutritional oil can include from 50 mg / g to 200 mg / g, from 60 mg / g to 150 mg / g, or from 70 mg / g to 130 mg / g palmitic acid based on the total weight of the nutritional oil.

[0099] In some examples, the nutritional oil can further include an additive. Non-limiting examples of additives can include an antioxidant, an antimicrobial agent, a flavoring, an odormasking agent, an edible oil and / or biofuel, the like, or combinations thereof.

[0100] A variety of added antioxidants can be included in the nutritional oils described herein. In some examples, the antioxidant can be or include a natural antioxidant, such as a tocopherol, a phospholipid, ascorbic acid / ascorbyl palmitate, phytic acid, a phenolic acid, citric acid, rosemary extract, the like, or a combination thereof. In some additional examples, the antioxidant can be or include a synthetic antioxidant, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, tertiary butyl hydroquinone (TBHQ), the like, or a combination thereof. In some specific examples, the antioxidant in the oil composition can be or include a tocopherol, ascorbyl palmitate, rosemary extract, or a combination thereof.

[0101] In some specific examples, the nutritional oils described herein can include greater than or equal to 50 ppm of an antioxidant based on the total weight of the nutritional oil. In still additional examples, the nutritional oils described herein can include greater than or equal to 75 ppm antioxidant, greater than or equal to 100 ppm antioxidant, greater than or equal to 125 ppm antioxidant, or greater than or equal to 150 ppm antioxidant based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include greater than or equal to 300 ppm antioxidant, greater than or equal to 500 ppm antioxidant, greater than or equal to 700 ppm antioxidant, greater than or equal to 900 ppm antioxidant, greater than or equal to1200 ppm antioxidant, greater than or equal to 1500 ppm antioxidant, greater than or equal to1800 ppm antioxidant, greater than or equal to 2000 ppm antioxidant, greater than or equal to2200 ppm antioxidant, greater than or equal to 2400 ppm antioxidant, greater than or equal to2600 ppm antioxidant, greater than or equal to 2800 ppm antioxidant, or greater than or equal to3000 ppm antioxidant based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include less than or equal to 10000 ppm antioxidant, less than or equal to 7500 ppm antioxidant, less than or equal to 5000 ppm antioxidant, less than or equal to 4000 ppm antioxidant, less than or equal to 3000 ppm antioxidant, or less than or equal to 2500 ppm antioxidant based on the total weight of the nutritional oil. In still additional examples, the nutritional oils can include less than or equal to 500 ppm antioxidant, less than or equal to 400 ppm antioxidant, less than or equal to 300 ppm antioxidant, less than or equal to 200 ppm antioxidant based on the total weight of the nutritional oil.

[0102] In some examples, the nutritional oil can include low levels of 3-monochloropropane-1 ,2- diol (3-MCPD) and 2, 3-epoxy-1 -propanol (glycidol). When referring to 3-MCPD and / or glycidol, such references also include reference to their respective esters unless otherwise specified. In some examples, the nutritional oils can include less than 1 ppm 3-MCPD based on the total weight of the nutritional oil. In some additional examples, the nutritional oils can include less than 0.5 ppm, less than 0.4 ppm, less than 0.3 ppm, less than 0.2 ppm, less than 0.15 ppm, or less than 0.1 ppm 3-MCPD based on the total weight of the nutritional oil.

[0103] In some examples, the nutritional oil can include less than 0.5 ppm glycidol based on the total weight of the nutritional oil. In some additional examples, the nutritional oil can include less than 0.2 ppm, less than 0.1 ppm, less than 0.05 ppm, less than 0.04 ppm, less than 0.03 ppm, or less than 0.02 ppm glycidol based on the total weight of the nutritional oil.

[0104] The amount of 3-MCPD and glycidol can be determined using any suitable analytical methodology. In some non-limiting examples, 3-MCPD content and glycidol content can be determined using GC-MS / MS.

[0105] The nutritional oil can typically have a low anisidine value (AV). In some examples, the nutritional oil can have an AV of less than 20. In still additional examples, the nutritional oil can have an AV of less than 15, less than 12, less than 10, less than 8, or less than 5. Anisidine value can typically be determined using AOCS Cd 18-90.

[0106] The nutritional oil can also typically have a low peroxide value (PV). In some examples, the nutritional oil can have a PV of less than 10 meq / kg based on the total weight of the nutritional oil. In still additional examples, the nutritional oil can have a PV of less than 8 meq / kg, less than 5 meq / kg, less than 3 meq / kg, less than 2 meq / kg, or less than 1 meq / kg based on thetotal weight of the nutritional oil. Peroxide value can typically be determined using AOCS Cd 8- 53.

[0107] The nutritional oils described herein can be formulated into a variety of nutritional or pharmaceutical products to exploit their biological or nutritional properties. Non-limiting examples of potential applications can include pharmaceuticals, food supplements, animal feed additives, cosmetics, and the like.

[0108] The nutritional oils described herein can be incorporated into a variety of food or feed supplements. For example, the nutritional oils can be included in a variety of beverages such as milk, sports drinks, energy drinks, teas, juices, etc. The nutritional oils can also be incorporated into confections, such as jellies and biscuits, dairy products, processed food products, such as soft rice or porridge, infant formula, breakfast cereals, and a variety of other food products.Other non-limiting examples can include pet foods, such as cat foods, dog foods, poultry foods, fish foods (including aquaculture), etc.

[0109] In additional examples, the nutritional oils can be incorporated into a dietary supplement, such as a multivitamin, an omega fatty acid supplement, or other suitable dietary supplement.

[0110] In some examples, the nutritional oils can be incorporated into a pharmaceutical composition. The pharmaceutical composition can be formulated with a pharmaceutically acceptable carrier to be administered to a subject as a suitable dosage form. In some examples, the pharmaceutical composition can be formulated as a liquid dosage form, such as a suspension, an elixir, an oil, a gel, a paste, or the like. In some additional examples, the pharmaceutical composition can be formulated as a solid dosage form, such as a powder, a tablet, a capsule, a gummy, a lozenge, or the like.

[0111] The nutritional oils can be used in a variety of other nutritional and / or pharmaceutical applications for humans and animals in addition to those specifically referenced herein.Nutritional Oil Blends

[0112] In some examples, the nutritional oils described herein can be blended with other nutritional oils to form a nutritional oil blend. In some examples, the blend can include two microbial oils. In some examples, the blend can include a microbial oil and a fish oil. In some examples, the blend can include a microbial oil and plant oil. In some examples, the blend can include a plant oil and a fish oil. In some examples, the blend can include two plant oils.

[0113] In some specific examples, the nutritional oil blend can include a first nutritional oil including ARA blended with a second nutritional oil including DHA. The first nutritional oil and the second nutritional oil can be blended together at a variety of ratios to form a nutritional oilblend having a variety of ARA / DHA ratios. In some examples, the nutritional oil blend can include ARA and DHA at a weight ratio of from 0.1 to 5 ARA / DHA. In still additional examples, the nutritional oil blend can include ARA and DHA at a weight ratio of from 0.5 to 3. In yet additional examples, the nutritional oil blend can include ARA and DHA at a weight ratio of from 0.1 to 1 , from 0.5 to 1.5, from 1 to 2, from 1.5 to 2.5, from 2 to 3, or from 2.5 to 3.5 ARA / DHA.

[0114] In some specific examples, the first nutritional oil comprising ARA can be produced from fungi of the order Mortierellales. In some specific examples, the second nutritional oil comprising DHA can be produced from a microalgae of the order Thraustochytriales (e.g., microorganisms of the genus Thraustochytrium or Schizochytrium). In some specific examples, the second nutritional oil comprising DHA can be produced from a dinoflagellate microalgae (e.g., Crypthecodinium cohnii).

[0115] The nutritional oil blends described herein can typically have less than or equal to 6 ppm MOSH based on the total weight of the nutritional oil blends. In some additional examples, the nutritional oil blends described herein can include less than or equal to 5 ppm MOSH, or less than or equal to 3 ppm MOSH based on the total weight of the nutritional oil blend. In some specific examples, the nutritional oil blends described herein can include MOSH in an amount less than or equal to 6 ppm, less than or equal to 5 ppm, less than or equal to 4 ppm, less than or equal to 3 ppm, less than or equal to 2.5 ppm, less than or equal to 2 ppm, less than or equal to 1 .5 ppm, less than or equal to 1 ppm, or less than or equal to 0.5 ppm based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include MOSH in an amount from 0.1 ppm to 6 ppm based on the total weight of the nutritional oil blend. In some further examples, the nutritional oil can include MOSH in an amount from 0.1 ppm to 2 ppm, from 0.5 ppm to 3 ppm, 1 ppm to 4 ppm, or 1.5 ppm to 5 ppm based on the total weight of the nutritional oil blend.

[0116] In some specific examples, the nutritional oil blends described herein can include MOAH in an amount less than or equal to 5 ppm, less than or equal to 4 ppm, less than or equal to 3 ppm, less than or equal to 2 ppm, less than or equal to 1.5 ppm, less than or equal to 1 ppm, or less than or equal to 0.5 ppm based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends described herein can include MOAH in an amount from 0.001 ppm to 2 ppm or 0.01 to 5 ppm based on the total weight of the nutritional oil blend.

[0117] MOSH and MOAH can be detected and measured using any suitable analytical methodology. As a non-limiting example, MOSH and MOAH can be isolated using solid-phase extraction technology and subsequently analyzed using gas chromatography with flameionization detection. MOSH and MOAH can also be analyzed using DIN EN 16995: 2017-08, or similar.

[0118] In some further examples, the nutritional oil blends can include ARA in an amount greater than or equal to 100 mg / g based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include ARA in an amount greater than or equal to 150 mg / g, greater than or equal to 200 mg / g, greater than or equal to 250 mg / g, greater than or equal to 300 mg / g, or greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend. In still additional examples, the nutritional oil blends can include ARA in an amount from 100 mg / g to 200 mg / g, from 150 mg / g to 250 mg / g, from 200 mg / g to 300 mg / g, from 250 mg / g to 350 mg / g, or from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0119] In some further examples, the nutritional oil blends can include DHA in an amount greater than or equal to 100 mg / g based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include DHA in an amount greater than or equal to 150 mg / g, greater than or equal to 200 mg / g, greater than or equal to 250 mg / g, greater than or equal to 300 mg / g, or greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend. In still additional examples, the nutritional oil blends can include DHA in an amount from 100 mg / g to 200 mg / g, from 150 mg / g to 250 mg / g, from 200 mg / g to 300 mg / g, from 250 mg / g to 350 mg / g, or from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0120] In some examples, the nutritional oil blends can further include an additive. Non-limiting examples of additives can include an antioxidant, an antimicrobial agent, a flavoring, an odormasking agent, an edible oil and / or biofuel, the like, or combinations thereof.

[0121] A variety of added antioxidants can be included in the nutritional oil blends described herein. In some examples, the antioxidant can be or include a natural antioxidant, such as a tocopherol, a phospholipid, ascorbic acid / ascorbyl palmitate, phytic acid, a phenolic acid, citric acid, rosemary extract, the like, or a combination thereof. In some additional examples, the antioxidant can be or include a synthetic antioxidant, such as butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, tertiary butyl hydroquinone (TBHQ), the like, or a combination thereof. In some specific examples, the antioxidant in the oil composition can be or include a tocopherol, ascorbyl palmitate, rosemary extract, or a combination thereof.

[0122] In some specific examples, the nutritional oil blends described herein can include greater than or equal to 50 ppm of an antioxidant based on the total weight of the nutritional oil blend. In still additional examples, the nutritional oil blends described herein can include greater than orequal to 75 ppm antioxidant, greater than or equal to 100 ppm antioxidant, greater than or equal to 125 ppm antioxidant, or greater than or equal to 150 ppm antioxidant based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include greater than or equal to 300 ppm antioxidant, greater than or equal to 500 ppm antioxidant, greater than or equal to 700 ppm antioxidant, greater than or equal to 900 ppm antioxidant, greater than or equal to 1200 ppm antioxidant, greater than or equal to 1500 ppm antioxidant, greater than or equal to 1800 ppm antioxidant, greater than or equal to 2000 ppm antioxidant, greater than or equal to 2200 ppm antioxidant, greater than or equal to 2400 ppm antioxidant, greater than or equal to 2600 ppm antioxidant, greater than or equal to 2800 ppm antioxidant, or greater than or equal to 3000 ppm antioxidant based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include less than or equal to 10000 ppm antioxidant, less than or equal to 7500 ppm antioxidant, less than or equal to 5000 ppm antioxidant, less than or equal to 4000 ppm antioxidant, less than or equal to 3000 ppm antioxidant, or less than or equal to 2500 ppm antioxidant based on the total weight of the nutritional oil blend. In still additional examples, the nutritional oil blends can include less than or equal to 500 ppm antioxidant, less than or equal to 400 ppm antioxidant, less than or equal to 300 ppm antioxidant, less than or equal to 200 ppm antioxidant based on the total weight of the nutritional oil blend.

[0123] In some examples, the nutritional oil blend can include low levels of 3- monochloropropane-1,2-diol (3-MCPD) and 2, 3-epoxy-1 -propanol (glycidol). When referring to 3-MCPD and / or glycidol, such references also include reference to their respective esters unless otherwise specified. In some examples, the nutritional oil blends can include less than 1 ppm 3-MCPD based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include less than 0.5 ppm, less than 0.4 ppm, less than 0.3 ppm, less than 0.2 ppm, less than 0.15 ppm, or less than 0.1 ppm 3-MCPD based on the total weight of the nutritional oil blend.

[0124] In some examples, the nutritional oil blends can include less than 0.5 ppm glycidol based on the total weight of the nutritional oil blend. In some additional examples, the nutritional oil blends can include less than 0.2 ppm, less than 0.1 ppm, less than 0.05 ppm, less than 0.04 ppm, less than 0.03 ppm, or less than 0.02 ppm glycidol based on the total weight of the nutritional oil blend.

[0125] The amount of 3-MCPD and glycidol can be determined using any suitable analytical methodology. In some non-limiting examples, 3-MCPD content and glycidol content can be determined using GC-MS / MS.

[0126] The nutritional oil blends can typically have a low anisidine value (AV). In some examples, the nutritional oil blends can have an AV of less than 20. In still additional examples, the nutritional oil can have an AV of less than 15, less than 12, less than 10, less than 8, or less than 5. Anisidine value can typically be determined using AOCS Cd 18-90.

[0127] The nutritional oil can also typically have a low peroxide value (PV). In some examples, the nutritional oil blends can have a PV of less than 10 meq / kg based on the total weight of the nutritional oil blend. In still additional examples, the nutritional oil blends can have a PV of less than 8 meq / kg, less than 5 meq / kg, less than 3 meq / kg, less than 2 meq / kg, or less than 1 meq / kg based on the total weight of the nutritional oil blend. Peroxide value can typically be determined using AOCS Cd 8-53.

[0128] The nutritional oil blends described herein can be formulated into a variety of nutritional or pharmaceutical products to exploit their biological or nutritional properties. Non-limiting examples of potential applications can include pharmaceuticals, food supplements, animal feed additives, cosmetics, and the like.

[0129] The nutritional oil blends described herein can be incorporated into a variety of food or feed supplements. For example, the nutritional oil blends can be included in a variety of beverages such as milk, sports drinks, energy drinks, teas, juices, etc. The nutritional oil blends can also be incorporated into confections, such as jellies and biscuits, dairy products, processed food products, such as soft rice or porridge, infant formula, breakfast cereals, and a variety of other food products. Other non-limiting examples can include pet foods, such as cat foods, dog foods, poultry foods, fish foods (including aquaculture), etc.

[0130] In additional examples, the nutritional oil blends can be incorporated into a dietary supplement, such as a multivitamin, an omega fatty acid supplement, or other suitable dietary supplement.

[0131] In some examples, the nutritional oil blends can be incorporated into a pharmaceutical composition. The pharmaceutical composition can be formulated with a pharmaceutically acceptable carrier to be administered to a subject as a suitable dosage form. In some examples, the pharmaceutical composition can be formulated as a liquid dosage form, such as a suspension, an elixir, an oil, a gel, a paste, or the like. In some additional examples, the pharmaceutical composition can be formulated as a solid dosage form, such as a powder, a tablet, a capsule, a gummy, a lozenge, or the like.

[0132] The nutritional oil blends can be used in a variety of other nutritional and / or pharmaceutical applications for humans and animals in addition to those specifically referenced herein.Example Embodiments

[0133] Various compositions and methods are disclosed herein. To aid in the understanding of these compositions and methods, a number of non-limiting example embodiments are provided in the following numbered clauses for illustration purposes only. The following example embodiments can be combined together in any suitable combination, unless the context clearly suggests otherwise.

[0134] Clause 1. A nutritional oil, comprising:ARA in an amount greater than or equal to 300 mg / g based on a total weight of the nutritional oil; free fatty acids in an amount greater than or equal to 0.1 wt% based on the total weight of the nutritional oil; and less than 7 ppm MOSH based on the total weight of the nutritional oil.

[0135] Clause 2. The nutritional oil of clause 1 , comprising ARA in an amount greater than or equal to 350 mg / g, optionally greater than or equal to 400 mg / g, optionally greater than or equal to 450 mg / g, or optionally greater than or equal to 500 mg / g, or optionally greater than or equal to 550 mg / g based on the total weight of the nutritional oil.

[0136] Clause 3. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 400 mg / g based on the total weight of the nutritional oil.

[0137] Clause 4. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 450 mg / g based on the total weight of the nutritional oil.

[0138] Clause 5. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 500 mg / g based on the total weight of the nutritional oil.

[0139] Clause 6. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 550 mg / g based on the total weight of the nutritional oil.

[0140] Clause 7. The nutritional oil of any one of the preceding clauses, comprising ARA in an amount from 300 mg / g to 400 mg / g, optionally from 350 mg / g to 450 mg / g, optionally from 400 mg / g to 500 mg / g, optionally from 450 mg / g to 550 mg / g, or optionally from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0141] Clause 8. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 350 mg / g to 450 mg / g based on the total weight of the nutritional oil.

[0142] Clause 9. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 400 mg / g to 500 mg / g based on the total weight of the nutritional oil.

[0143] Clause 10. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 450 mg / g to 550 mg / g based on the total weight of the nutritional oil.

[0144] Clause 11. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0145] Clause 12. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount less than or equal to 6 ppm, optionally less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3.5 ppm based on the total weight of the nutritional oil.

[0146] Clause 13. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount less than or equal to 5 ppm based on the total weight of the nutritional oil.

[0147] Clause 14. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0148] Clause 15. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount less than or equal to 3.5 ppm based on the total weight of the nutritional oil.

[0149] Clause 16. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount from 0.5 ppm to less than 7 ppm based on the total weight of the nutritional oil.

[0150] Clause 17. The nutritional oil of any one of the preceding clauses, comprising MOSH in an amount from 2 ppm to less than 7 ppm.

[0151] Clause 18. The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1 .5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0152] Clause 19. The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0153] Clause 20. The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil.

[0154] Clause 21 . The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0155] Clause 22. The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 1 .5 ppm based on the total weight of the nutritional oil.

[0156] Clause 23. The nutritional oil of any one of the preceding clauses, comprising MOAH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0157] Clause 24. The nutritional oil of any one of the preceding clauses, further comprising an added antioxidant. 1

[0158] Clause 25. The nutritional oil of clause 24, wherein the added antioxidant comprises a tocopherol.

[0159] Clause 26. The nutritional oil of any one of the preceding clauses, comprising 50 wt% or greater, 55 wt% or greater, 60 wt% or greater, 65 wt% or greater, 70 wt% or greater, 75 wt% or greater, 80 wt% or greater, or 85 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0160] Clause 27. The nutritional oil of any one of the preceding clauses, comprising 50 wt% or greater, preferably 55 wt% or greater, preferably 60 wt% or greater, preferably 65 wt% or greater, more preferably 70 wt% or greater, more preferably 75 wt% or greater, more preferably 80 wt% or greater, or more preferably 85 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0161] Clause 28. The nutritional oil of any one of the preceding clauses, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, less than or equal to 0.15 ppm, or less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0162] Clause 29. The nutritional oil of any one of the preceding clauses, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, preferably less than or equal to 0.15 ppm, or more preferably less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0163] Clause 30. The nutritional oil of any one of the preceding clauses, comprising glycidol in an amount less than or equal to 0.1 ppm, less than or equal to 0.05 ppm, less than or equal to 0.04 ppm, less than or equal to 0.03 ppm, or less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0164] Clause 31. The nutritional oil of any one of the preceding clauses, comprising glycidol in an amount less than or equal to 0.1 ppm, preferably less than or equal to 0.05 ppm, preferably less than or equal to 0.04 ppm, more preferably less than or equal to 0.03 ppm, or more preferably less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0165] Clause 32. The nutritional oil of any one of the preceding clauses, wherein the nutritional oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

[0166] Clause 33. The nutritional oil of any one of the preceding clauses, wherein the nutritional oil has an anisidine value of less than 20, preferably less than 15, preferably less than 12, more preferably less than 10, or more preferably less than 8.

[0167] Clause 34. The nutritional oil of any one of the preceding clauses, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, less than or equal to 8 meq / kg, lessthan or equal to 5 meq / kg, less than or equal to 3 meq / kg, less than or equal to 2 meq / kg, or less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0168] Clause 35. The nutritional oil of any one of the preceding clauses, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, preferably less than or equal to 8 meq / kg, preferably less than or equal to 5 meq / kg, preferably less than or equal to 3 meq / kg, more preferably less than or equal to 2 meq / kg, or more preferably less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0169] Clause 36. The nutritional oil of any one of the preceding clauses, wherein the nutritional oil is a microbial oil.

[0170] Clause 37. The nutritional oil of clause 36, wherein the microbial oil is produced by Mortierella alpina.

[0171] Clause 38. A nutritional oil, comprising:ARA in an amount greater than or equal to 300 mg / g based on a total weight of the nutritional oil; less than 0.1 wt% free fatty acids based on the total weight of the nutritional oil; and less than or equal to 3 ppm MOSH based on the total weight of the nutritional oil.

[0172] Clause 39. The nutritional oil of clause 38, comprising ARA in an amount greater than or equal to 350 mg / g, optionally greater than or equal to 400 mg / g, optionally greater than or equal to 450 mg / g, or optionally greater than or equal to 500 mg / g based on the total weight of the nutritional oil.

[0173] Clause 40. The nutritional oil of any one of clauses 38 to 39, comprising ARA in an amount greater than or equal to 400 mg / g based on the total weight of the nutritional oil.

[0174] Clause 41 . The nutritional oil of any one of clauses 38 to 40, comprising ARA in an amount greater than or equal to 450 mg / g based on the total weight of the nutritional oil.

[0175] Clause 42. The nutritional oil of any one of clauses 38 to 41, comprising ARA in an amount greater than or equal to 500 mg / g based on the total weight of the nutritional oil.

[0176] Clause 43. The nutritional oil of any one of clauses 38 to 42, comprising ARA in an amount greater than or equal to 550 mg / g based on the total weight of the nutritional oil.

[0177] Clause 44. The nutritional oil of any one of clauses 38 to 43, comprising ARA in an amount from 300 mg / g to 400 mg / g, optionally from 350 mg / g to 450 mg / g, or optionally from 400 mg / g to 500 mg / g based on the total weight of the nutritional oil.

[0178] Clause 45. The nutritional oil of any one of clauses 38 to 44, comprising ARA in an amount from 350 mg / g to 450 mg / g based on the total weight of the nutritional oil.

[0179] Clause 46. The nutritional oil of any one of clauses 38 to 44, comprising ARA in an amount from 400 mg / g to 500 mg / g based on the total weight of the nutritional oil.

[0180] Clause 47. The nutritional oil of any one of clauses 38 to 44, comprising ARA in an amount from 450 mg / g to 550 mg / g based on the total weight of the nutritional oil.

[0181] Clause 48. The nutritional oil of any one of clauses 38 to 44, comprising ARA in an amount from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0182] Clause 49. The nutritional oil of any one of clauses 38 to 48, comprising MOSH in an amount less than or equal to 2.5 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1.5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0183] Clause 50. The nutritional oil of any one of clauses 38 to 49, comprising MOSH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0184] Clause 51 . The nutritional oil of any one of clauses 38 to 50, comprising MOSH in an amount less than or equal to 1.5 ppm based on the total weight of the nutritional oil.

[0185] Clause 52. The nutritional oil of any one of clauses 38 to 51, comprising MOSH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0186] Clause 53. The nutritional oil of any one of clauses 38 to 52, comprising MOSH in an amount from 0.5 ppm to 2 ppm based on the total weight of the nutritional oil.

[0187] Clause 54. The nutritional oil of any one of clauses 38 to 53, comprising MOSH in an amount from 1 ppm to 3 ppm.

[0188] Clause 55. The nutritional oil of any one of clauses 38 to 54, comprising MOAH in an amount less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1.5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0189] Clause 56. The nutritional oil of any one of clauses 38 to 55, comprising MOAH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0190] Clause 57. The nutritional oil of any one of clauses 38 to 56, comprising MOAH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil.

[0191] Clause 58. The nutritional oil of any one of clauses 38 to 57, comprising MOAH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0192] Clause 59. The nutritional oil of any one of clauses 38 to 58, comprising MOAH in an amount less than or equal to 1.5 ppm based on the total weight of the nutritional oil.

[0193] Clause 60. The nutritional oil of any one of clauses 38 to 59, comprising MOAH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0194] Clause 61. The nutritional oil of any one of clauses 38 to 60, further comprising an added antioxidant.

[0195] Clause 62. The nutritional oil of clauses 61 , wherein the added antioxidant comprises a tocopherol.

[0196] Clause 63. The nutritional oil of any one of clauses 38 to 62, comprising 60 wt% or greater, 65 wt% or greater, 70 wt% or greater, 75 wt% or greater, 80 wt% or greater, 85 wt% or greater, 90 wt% or greater, or 95 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0197] Clause 64. The nutritional oil of any one of clauses 38 to 63, comprising 50 wt% or greater, preferably 55 wt% or greater, preferably 60 wt% or greater, preferably 65 wt% or greater, more preferably 70 wt% or greater, more preferably 75 wt% or greater, more preferably 80 wt% or greater, or more preferably 85 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0198] Clause 65. The nutritional oil of any one of clauses 38 to 64, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, less than or equal to 0.15 ppm, or less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0199] Clause 66. The nutritional oil of any one of clauses 38 to 65, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, preferably less than or equal to 0.15 ppm, or more preferably less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0200] Clause 67. The nutritional oil of any one of clauses 38 to 66, comprising glycidol in an amount less than or equal to 0.1 ppm, less than or equal to 0.05 ppm, less than or equal to 0.04 ppm, less than or equal to 0.03 ppm, or less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0201] Clause 68. The nutritional oil of any one of clauses 38 to 67, comprising glycidol in an amount less than or equal to 0.1 ppm, preferably less than or equal to 0.05 ppm, preferably less than or equal to 0.04 ppm, more preferably less than or equal to 0.03 ppm, or more preferably less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0202] Clause 69. The nutritional oil of any one of clauses 38 to 68, wherein the nutritional oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

[0203] Clause 70. The nutritional oil of any one of clauses 38 to 69, wherein the nutritional oil has an anisidine value of less than 20, preferably less than 15, preferably less than 12, more preferably less than 10, or more preferably less than 8.

[0204] Clause 71 . The nutritional oil of any one of clauses 38 to 70, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, less than or equal to 8 meq / kg, lessthan or equal to 5 meq / kg, less than or equal to 3 meq / kg, less than or equal to 2 meq / kg, or less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0205] Clause 72. The nutritional oil of any one of clauses 38 to 71, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, preferably less than or equal to 8 meq / kg, preferably less than or equal to 5 meq / kg, preferably less than or equal to 3 meq / kg, more preferably less than or equal to 2 meq / kg, or more preferably less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0206] Clause 73. The nutritional oil of any one of clauses 38 to 72, wherein the nutritional oil is a microbial oil.

[0207] Clause 74. The nutritional oil of clause 73, wherein the microbial oil is produced by Mortierella alpina.

[0208] Clause 75. A nutritional oil blend, comprising: a first nutritional oil comprising ARA blended with a second nutritional oil comprising DHA, wherein the nutritional oil blend comprises ARA and DHA at a weight ratio of from 0.5 to 3 ARA / DHA, and wherein the nutritional oil blend comprises MOSH in an amount less than or equal to 4 ppm.

[0209] Clause 76. The nutritional oil blend according to clause 75, comprising ARA in an amount greater than or equal to 100 mg / g, optionally greater than or equal to 150 mg / g, optionally greater than or equal to 200 mg / g, optionally greater than or equal to 250 mg / g, optionally greater than or equal to 300 mg / g, optionally greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend.

[0210] Clause 77. The nutritional oil blend according to any one of clauses 75 to 76, comprising ARA in an amount greater than or equal to 150 mg / g based on the total weight of the nutritional oil blend.

[0211] Clause 78. The nutritional oil blend according to any one of clauses 75 to 77, comprising ARA in an amount greater than or equal to 200 mg / g based on the total weight of the nutritional oil blend.

[0212] Clause 79. The nutritional oil blend according to any one of clauses 75 to 78, comprising ARA in an amount greater than or equal to 250 mg / g based on the total weight of the nutritional oil blend.

[0213] Clause 80. The nutritional oil blend according to any one of clauses 75 to 79, comprising ARA in an amount greater than or equal to 300 mg / g based on the total weight of the nutritional oil blend.

[0214] Clause 81. The nutritional oil blend according to any one of clauses 75 to 80, comprising ARA in an amount greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend.

[0215] Clause 82. The nutritional oil blend according to clause 75, comprising ARA in an amount from 100 mg / g to 200 mg / g, optionally from 150 mg / g to 250 mg / g, optionally from 200 mg / g to 300 mg / g, optionally from 250 mg / g to 350 mg / g, or optionally from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0216] Clause 83. The nutritional oil blend according to clause 75, comprising ARA in an amount from 150 mg / g to 250 mg / g based on the total weight of the nutritional oil blend.

[0217] Clause 84. The nutritional oil blend according to clause 75, comprising ARA in an amount from 200 mg / g to 300 mg / g based on the total weight of the nutritional oil blend.

[0218] Clause 85. The nutritional oil blend according to clause 75, comprising ARA in an amount from 250 mg / g to 350 mg / g based on the total weight of the nutritional oil blend.

[0219] Clause 86. The nutritional oil blend according to clause 75, comprising ARA in an amount from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0220] Clause 87. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount greater than or equal to 100 mg / g, optionally greater than or equal to 150 mg / g, optionally greater than or equal to 200 mg / g, optionally greater than or equal to 250 mg / g, optionally greater than or equal to 300 mg / g, optionally greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend.

[0221] Clause 88. The nutritional oil blend according to any one of clauses 75 to 87, comprising DHA in an amount greater than or equal to 150 mg / g based on the total weight of the nutritional oil blend.

[0222] Clause 89. The nutritional oil blend according to any one of clauses 75 to 88, comprising DHA in an amount greater than or equal to 200 mg / g based on the total weight of the nutritional oil blend.

[0223] Clause 90. The nutritional oil blend according to any one of clauses 75 to 89, comprising DHA in an amount greater than or equal to 250 mg / g based on the total weight of the nutritional oil blend.

[0224] Clause 91. The nutritional oil blend according to any one of clauses 75 to 90, comprising DHA in an amount greater than or equal to 300 mg / g based on the total weight of the nutritional oil blend.

[0225] Clause 92. The nutritional oil blend according to any one of clauses 75 to 91, comprising DHA in an amount greater than or equal to 350 mg / g based on the total weight of the nutritional oil blend.

[0226] Clause 93. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount from 100 mg / g to 200 mg / g, optionally from 150 mg / g to 250 mg / g, optionally from 200 mg / g to 300 mg / g, optionally from 250 mg / g to 350 mg / g, or optionally from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0227] Clause 94. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount from 150 mg / g to 250 mg / g based on the total weight of the nutritional oil blend.

[0228] Clause 95. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount from 200 mg / g to 300 mg / g based on the total weight of the nutritional oil blend.

[0229] Clause 96. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount from 250 mg / g to 350 mg / g based on the total weight of the nutritional oil blend.

[0230] Clause 97. The nutritional oil blend according to any one of clauses 75 to 86, comprising DHA in an amount from 300 mg / g to 400 mg / g based on the total weight of the nutritional oil blend.

[0231] Clause 98. The nutritional oil blend according to any one of claims 75 to 97, comprising ARA and DHA at a weight ratio of from 0.5 to 3 ARA / DHA, optionally from 0.5 to 1.5 ARA / DHA, optionally from 1 to 2 ARA / DHA, optionally from 1.5 to 2.5 ARA / DHA, or optionally from 2 to 3 ARA / DHA.

[0232] Clause 99. The nutritional oil blend according to any one of claims 75 to 98, comprising ARA and DHA at a weight ratio of from 0.5 to 1.5 ARA / DHA.

[0233] Clause 100. The nutritional oil blend according to any one of claims 75 to 99, comprising ARA and DHA at a weight ratio of from 1 to 2 ARA / DHA.

[0234] Clause 101. The nutritional oil blend according to any one of claims 75 to 100, comprising ARA and DHA at a weight ratio of from 1 .5 to 2.5 ARA / DHA.

[0235] Clause 102. The nutritional oil blend according to any one of claims 75 to 101 , comprising ARA and DHA at a weight ratio of from 2 to 3 ARA / DHA.

[0236] Clause 103. The nutritional oil blend of any one of clauses 75 to 102, comprising MOSH in an amount less than or equal to 3.5 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2.5 ppm, optionally less than or equal to 2 ppm, optionally less than orequal to 1.5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil blend.

[0237] Clause 104. The nutritional oil blend of any one of clauses 75 to 103, comprising MOSH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil blend.

[0238] Clause 105. The nutritional oil blend of any one of clauses 75 to 104, comprising MOSH in an amount less than or equal to 2.5 ppm based on the total weight of the nutritional oil blend.

[0239] Clause 106. The nutritional oil blend of any one of clauses 75 to 105, comprising MOSH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil blend.

[0240] Clause 107. The nutritional oil blend of any one of clauses 75 to 106, comprising MOSH in an amount less than or equal to 1.5 ppm based on the total weight of the nutritional oil blend.

[0241] Clause 108. The nutritional oil blend of any one of clauses 75 to 107, comprising MOSH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil blend.

[0242] Clause 109. The nutritional oil blend of any one of clauses 75 to 108, comprising MOSH in an amount from 0.5 ppm to 3 ppm based on the total weight of the nutritional oil blend.

[0243] Clause 110. The nutritional oil blend of any one of clauses 75 to 109, comprising MOSH in an amount from 1 ppm to 4 ppm based on the total weight of the nutritional oil blend.

[0244] Clause 111. The nutritional oil blend of any one of clauses 75 to 110, comprising MOAH in an amount less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1.5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil blend.

[0245] Clause 112. The nutritional oil blend of any one of clauses 75 to 111 , comprising MOAH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil blend.

[0246] Clause 113. The nutritional oil blend of any one of clauses 75 to 112, comprising MOAH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil blend.

[0247] Clause 114. The nutritional oil blend of any one of clauses 75 to 113, comprising MOAH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil blend.

[0248] Clause 115. The nutritional oil blend of any one of clauses 75 to 114, comprising MOAH in an amount less than or equal to 1 .5 ppm based on the total weight of the nutritional oil blend.

[0249] Clause 116. The nutritional oil blend of any one of clauses 75 to 115, comprising MOAH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil blend.

[0250] Clause 117. The nutritional oil blend of any one of clauses 75 to 116, further comprising an added antioxidant.

[0251] Clause 118. The nutritional oil blend of claim 117, wherein the added antioxidant comprises a tocopherol.

[0252] Clause 119. The nutritional oil blend of any one of claims 75 to 118, comprising 60 wt% or greater, 65 wt% or greater, 70 wt% or greater, 75 wt% or greater, 80 wt% or greater, 85 wt% or greater, 90 wt% or greater, or 95 wt% or greater of triglycerides based on the total weight of the nutritional oil blend.

[0253] Clause 120. The nutritional oil blend of any one of claims 75 to 119, comprising 60 wt% or greater, preferably 65 wt% or greater, more preferably 70 wt% or greater, more preferably 75 wt% or greater, more preferably 80 wt% or greater, more preferably 85 wt% or greater, more preferably 90 wt% or greater, or more preferably 95 wt% or greater of triglycerides based on the total weight of the nutritional oil blend.

[0254] Clause 121. The nutritional oil blend of any one of claims 75 to 120, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, less than or equal to 0.15 ppm, or less than or equal to 0.1 ppm based on the total amount of nutritional oil blend.

[0255] Clause 122. The nutritional oil blend of any one of claims 75 to 121 , comprising 3-MCPD in an amount less than or equal to 0.2 ppm, preferably less than or equal to 0.15 ppm, or more preferably less than or equal to 0.1 ppm based on the total amount of nutritional oil blend.

[0256] Clause 123. The nutritional oil blend of any one of claims 75 to 122, comprising glycidol in an amount less than or equal to 0.1 ppm, less than or equal to 0.05 ppm, less than or equal to 0.04 ppm, less than or equal to 0.03 ppm, or less than or equal to 0.02 ppm based on the total amount of nutritional oil blend.

[0257] Clause 124. The nutritional oil blend of any one of claims 75 to 123, comprising glycidol in an amount less than or equal to 0.1 ppm, preferably less than or equal to 0.05 ppm, preferably less than or equal to 0.04 ppm, more preferably less than or equal to 0.03 ppm, or more preferably less than or equal to 0.02 ppm based on the total amount of nutritional oil blend.

[0258] Clause 125. The nutritional oil blend of any one of claims 75 to 124, wherein the nutritional oil blend has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

[0259] Clause 126. The nutritional oil blend of any one of claims 75 to 125, wherein the nutritional oil blend has an anisidine value of less than 20, preferably less than 15, preferably less than 12, more preferably less than 10, or more preferably less than 8.

[0260] Clause 127. The nutritional oil blend of any one of claims 75 to 126, wherein the nutritional oil blend has a peroxide value of less than or equal to 10 meq / kg, less than or equalto 8 meq / kg, less than or equal to 5 meq / kg, less than or equal to 3 meq / kg, less than or equal to 2 meq / kg, or less than or equal to 1 meq / kg based on the total weight of the nutritional oil blend.

[0261] Clause 128. The nutritional oil blend of any one of claims 75 to 127, wherein the nutritional oil blend has a peroxide value of less than or equal to 10 meq / kg, preferably less than or equal to 8 meq / kg, preferably less than or equal to 5 meq / kg, preferably less than or equal to 3 meq / kg, more preferably less than or equal to 2 meq / kg, or more preferably less than or equal to 1 meq / kg based on the total weight of the nutritional oil blend.

[0262] Clause 129. The nutritional oil blend of any one of clauses 75 to 128, wherein the nutritional oil blend comprises a microbial oil.

[0263] Clause 130. The nutritional oil blend of clause 129, wherein the microbial oil comprises DHA.

[0264] Clause 131. The nutritional oil blend of clause 130, wherein the microbial oil is produced by Crypthecodinium cohnii.

[0265] Clause 132. The nutritional oil blend of clause 130, wherein the microbial oil is produced by a microorganism of the genus Thraustochytrium or Schizochytrium.

[0266] Clause 133. The nutritional oil blend of clause 129, wherein the microbial oil comprises ARA.

[0267] Clause 134. The nutritional oil blend of clause 133, wherein microbial oil is produced by Mortierella alpina.

[0268] Clause 135. The nutritional oil blend of any one of clauses 128 to 134, comprising a first microbial oil comprising ARA and a second microbial oil comprising DHA.

[0269] Clause 136. A method of producing a nutritional oil according to any one of clauses 1 to 74, comprising: i. combining a LC-PUFA-containing oil with an organic solvent; ii. removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude nutritional oil; and iii. recovering at least a portion of the organic solvent removed in step ii using a recovery solvent, the recovery solvent comprising less than 50 wt% of mineral oil based on a total weight of the recovery solvent, wherein the nutritional oil has less than 7 ppm MOSH.

[0270] Clause 137. A method of producing a nutritional oil, comprising: combining a LC-PUFA-containing oil with an organic solvent;removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude nutritional oil; and recovering at least a portion of the organic solvent removed in step ii using a recovery solvent, the recovery solvent comprising less than 50 wt% of mineral oil based on a total weight of the recovery solvent.

[0271] Clause 138. The method of any one of clauses 136 to 137, wherein the organic solvent comprises hexane.

[0272] Clause 139. The method of any one of clauses 136 to 138, wherein the organic solvent comprises at least a portion of a recovered organic solvent previously combined with a second LC-PUFA-containing oil.

[0273] Clause 140. The method of clause 139, wherein the recovered organic solvent comprises less than 100 ppm MOSH, less than 75 ppm MOSH, less than 50 ppm MOSH, less than 25 ppm MOSH, less than 10 ppm MOSH, less than 5 ppm MOSH, or less than 1 ppm MOSH.

[0274] Clause 141. The method of any one of clauses 139 and 140, wherein the recovered organic solvent comprises less than 100 ppm MOSH, preferably less than 75 ppm MOSH, preferably less than 50 ppm MOSH, preferably less than 25 ppm MOSH, more preferably less than 10 ppm MOSH, more preferably less than 5 ppm MOSH, or more preferably less than 1 ppm MOSH.

[0275] Clause 142. The method of any one of clauses 136 to 141 , wherein combining the LC- PUFA-containing oil with an organic solvent comprises extracting a microbial oil from a microorganism using the organic solvent.

[0276] Clause 143. The method of clause 142, wherein the microorganism is of the order Mortierellales.

[0277] Clause 144. The method of clause 143, wherein the microorganism is Mortierella alpina.

[0278] Clause 145. The method of clause 142, wherein the microorganism is of the order Thraustochytriales.

[0279] Clause 146. The method of clause 145, wherein the microorganism is of the genus Thraustochytrium or Schizochytrium.

[0280] Clause 147. The method of clause 142, wherein the microorganism is a dinoflagellate.

[0281] Clause 148. The method of clause 147, wherein the microorganism is Crypthecodinium cohnii.

[0282] Clause 149. The method of any one of clauses 136 to 148, wherein the recovery solvent comprises less than 40 wt%, less than 30 wt%, less than 20 wt%, less than 10 wt%, or less than 5 wt% mineral oil based on the total weight of the recovery solvent

[0283] Clause 150. The method of any one of clauses 136 to 149, wherein the recovery solvent comprises less than 40 wt%, preferably less than 30 wt%, preferably less than 20 wt%, more preferably less than 10 wt%, or more preferably less than 5 wt% mineral oil based on the total weight of the recovery solvent.

[0284] Clause 151. The method of any one of clauses 136 to 149, wherein the recovery solvent comprises triglycerides.

[0285] Clause 152. The method of any one of clauses 136 to 151, wherein the recovery solvent comprises sunflower oil, medium chain triglycerides, canola oil, or a combination thereof.

[0286] Clause 153. The method of any one of clauses 136 to 152, wherein the nutritional oil is the crude nutritional oil.

[0287] Clause 154. The method of any one of clauses 136 to 153, wherein the nutritional oil is a final nutritional oil obtained by further processing the crude nutritional oil to obtain the final nutritional oil.

[0288] Clause 155. The method of any one of clauses 136 to 154, wherein the nutritional oil comprises one or more of ARA, DHA, or ERA in an amount greater than or equal to 300 mg / g based on the total amount of the nutritional oil.

[0289] Clause 156. The method of any one of clauses 136 to 155, wherein the nutritional oil has MOSH in an amount less than or equal to 6 ppm, optionally less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2.5 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1 .5 ppm, or optionally less than or equal to 1 ppm, or optionally less than or equal to 0.5 ppm based on the total weight of the nutritional oil.

[0290] Clause 157. The method of any one of clauses 136 to 156, wherein the nutritional oil has MOSH in an amount less than or equal to 5 ppm based on the total weight of the nutritional oil.

[0291] Clause 158. The method of any one of clauses 136 to 157, wherein the nutritional oil has MOSH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0292] Clause 159. The method of any one of clauses 136 to 158, wherein the nutritional oil has MOSH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil.

[0293] Clause 160. The method of any one of clauses 136 to 159, wherein the nutritional oil has MOSH in an amount less than or equal to 2.5 ppm based on the total weight of the nutritional oil.

[0294] Clause 161. The method of any one of clauses 136 to 160, wherein the nutritional oil has MOSH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0295] Clause 162. The method of any one of clauses 136 to 161 , wherein the nutritional oil has MOSH in an amount less than or equal to 1 .5 ppm based on the total weight of the nutritional oil.

[0296] Clause 163. The method of any one of clauses 136 to 162, wherein the nutritional oil has MOSH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0297] Clause 164. The method of any one of clauses 136 to 163, wherein the nutritional oil has MOSH in an amount less than or equal to 0.5 ppm based on the total weight of the nutritional oil.

[0298] Clause 165. The method of any one of clauses 136 to 156, wherein the nutritional oil includes MOSH in an amount from 0.1 ppm to 2 ppm based on the total weight of the nutritional oil.

[0299] Clause 166. The method of any one of clauses 136 to 156, wherein the nutritional oil includes MOSH in an amount from 0.5 ppm to 3 ppm based on the total weight of the nutritional oil.

[0300] Clause 167. The method of any one of clauses 136 to 156, wherein the nutritional oil includes MOSH in an amount from 1 ppm to 4 ppm based on the total weight of the nutritional oil.

[0301] Clause 168. The method of any one of clauses 136 to 156, wherein the nutritional oil includes MOSH in an amount from 1.5 ppm to 5 ppm based on the total weight of the nutritional oil.

[0302] Clause 169. The method of any one of clauses 136 to 168, wherein the nutritional oil has MOAH in an amount less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1 .5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0303] Clause 170. The method of cany one of clauses 136 to 169, wherein the nutritional oil has MOAH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0304] Clause 171. The method of cany one of clauses 136 to 170, wherein the nutritional oil has MOAH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil.

[0305] Clause 172. The method of cany one of clauses 136 to 171 , wherein the nutritional oil has MOAH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0306] Clause 173. The method of cany one of clauses 136 to 172, wherein the nutritional oil has MOAH in an amount less than or equal to 1 .5 ppm based on the total weight of the nutritional oil.

[0307] Clause 174. The method of cany one of clauses 136 to 173, wherein the nutritional oil has MOAH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0308] Clause 175. The method of any one of clauses 136 to 174, wherein the nutritional oil comprises free fatty acids in an amount greater than 0.1 wt%, optionally greater than 0.15 wt%, greater than 0.2 wt%, optionally greater than 0.25 wt%, optionally greater than 0.3 wt%, optionally greater than 0.35 wt%, optionally greater than 0.4 wt%, optionally greater than 0.45 wt%, optionally greater than 0.5 wt%, or optionally greater than 0.55 wt% based on the total weight of the nutritional oil. Clause 176. The method of any one of clauses 136 to 175, wherein the nutritional oil comprises free fatty acids in an amount less than 0.1 wt% or less than 0.05 wt% based on the total weight of the nutritional oil.

[0309] Clause 177. The method of any one of clauses 136 to 176, wherein the nutritional oil comprises free fatty acids in an amount less than 0.1 wt% or preferably less than 0.05 wt% based on the total weight of the nutritional oil.

[0310] Clause 178. The method of any one of clauses 136 to 177, further comprising employing an equipment material comprising less than or equal to 50 wt% mineral oil based on a total weight of the equipment material.

[0311] Clause 179. The method of clause 178, wherein the equipment material is an equipment lubricant.

[0312] Clause 180. The method of any one of clauses 178 and 179, wherein the equipment material comprises canola oil, sunflower oil, mixed chain triglycerides, or a combination thereof.

[0313] Clause 181. A nutritional oil produced by the method of any one of clauses 137 to 180, comprising:ARA in an amount greater than or equal to 300 mg / g based on a total weight of the nutritional oil; and from 0.5 ppm to 10 ppm MOSH based on the total weight of the nutritional oil.

[0314] Clause 182. The nutritional oil of clause 1, comprising ARA in an amount greater than or equal to 350 mg / g, optionally greater than or equal to 400 mg / g, optionally greater than or equalto 450 mg / g, or optionally greater than or equal to 500 mg / g, or optionally greater than or equal to 550 mg / g based on the total weight of the nutritional oil.

[0315] Clause 183. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 400 mg / g based on the total weight of the nutritional oil.

[0316] Clause 184. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 450 mg / g based on the total weight of the nutritional oil.

[0317] Clause 185. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 500 mg / g based on the total weight of the nutritional oil.

[0318] Clause 186. The nutritional oil of any of the preceding clauses, comprising ARA in an amount greater than or equal to 550 mg / g based on the total weight of the nutritional oil.

[0319] Clause 187. The nutritional oil of any one of the preceding clauses, comprising ARA in an amount from 300 mg / g to 400 mg / g, optionally from 350 mg / g to 450 mg / g, optionally from 400 mg / g to 500 mg / g, optionally from 450 mg / g to 550 mg / g, or optionally from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0320] Clause 188. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 350 mg / g to 450 mg / g based on the total weight of the nutritional oil.

[0321] Clause 189. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 400 mg / g to 500 mg / g based on the total weight of the nutritional oil.

[0322] Clause 190. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 450 mg / g to 550 mg / g based on the total weight of the nutritional oil.

[0323] Clause 191. The nutritional oil of any one of clauses 1 to 7, comprising ARA in an amount from 500 mg / g to 600 mg / g based on the total weight of the nutritional oil.

[0324] Clause 192. The nutritional oil of any one of clauses 181 to 191, comprising free fatty acids in an amount greater than or equal to 0.1 wt% based on the total weight of the nutritional oil.

[0325] Clause 193. The nutritional oil of clause 192, comprising MOSH in an amount less than or equal to 7 ppm, optionally less than or equal to 6 ppm, optionally less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3.5 ppm based on the total weight of the nutritional oil.

[0326] Clause 194. The nutritional oil of any one of clauses 192 to 193, comprising MOSH in an amount less than or equal to 6 ppm based on the total weight of the nutritional oil.

[0327] Clause 195. The nutritional oil of any one of clauses 192 to 194, comprising MOSH in an amount less than or equal to 5 ppm based on the total weight of the nutritional oil.

[0328] Clause 196. The nutritional oil of any one of clauses 192 to 195, comprising MOSH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0329] Clause 197. The nutritional oil of any one of clauses 192 to 196, comprising MOSH in an amount less than or equal to 3.5 ppm based on the total weight of the nutritional oil.

[0330] Clause 198. The nutritional oil of any one of clauses 192 to 197, comprising MOSH in an amount from 0.5 ppm to less than 7 ppm based on the total weight of the nutritional oil.

[0331] Clause 199. The nutritional oil of any one of clauses 192 to 198, comprising MOSH in an amount from 2 ppm to less than 7 ppm.

[0332] Clause 200. The nutritional oil of any one of the clauses of 181 to 191 , comprising free fatty acids in an amount less than 0.1 wt% based on the total weight of the nutritional oil.

[0333] Clause 201. The nutritional oil of clause 200, comprising MOSH in an amount from 0.1 ppm to 5 ppm, optionally from 0.1 ppm to 2 ppm, optionally from 0.5 ppm to 3 ppm, optionally 1 ppm to 4 ppm, or optionally 1.5 ppm to 5 ppm based on the total weight of the nutritional oil.

[0334] Clause 202. The nutritional oil of any one of clauses 200 to 201 , comprising MOSH in an amount from 0.1 ppm to 2 ppm based on the total weight of the nutritional oil.

[0335] Clause 203. The nutritional oil of any one of clauses 200 to 201, comprising MOSH in an amount from 0.5 ppm to 3 ppm based on the total weight of the nutritional oil.

[0336] Clause 204. The nutritional oil of any one of clauses 200 to 201, comprising MOSH in an amount from 1 ppm to 4 ppm based on the total weight of the nutritional oil.

[0337] Clause 205. The nutritional oil of any one of clauses 200 to 201, comprising MOSH in an amount from 1.5 ppm to 5 ppm based on the total weight of the nutritional oil.

[0338] Clause 206. The nutritional oil of any one of clauses 181 to 205, comprising MOAH in an amount less than or equal to 5 ppm, optionally less than or equal to 4 ppm, optionally less than or equal to 3 ppm, optionally less than or equal to 2 ppm, optionally less than or equal to 1.5 ppm, or optionally less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0339] Clause 207. The nutritional oil of any one of clauses 181 to 206, comprising MOAH in an amount less than or equal to 4 ppm based on the total weight of the nutritional oil.

[0340] Clause 208. The nutritional oil of any one of clauses 181 to 207, comprising MOAH in an amount less than or equal to 3 ppm based on the total weight of the nutritional oil.

[0341] Clause 209. The nutritional oil of any one of clauses 181 to 208, comprising MOAH in an amount less than or equal to 2 ppm based on the total weight of the nutritional oil.

[0342] Clause 210. The nutritional oil of any one of clauses 181 to 209, comprising MOAH in an amount less than or equal to 1.5 ppm based on the total weight of the nutritional oil.

[0343] Clause 211. The nutritional oil of any one of clauses 181 to 210, comprising MOAH in an amount less than or equal to 1 ppm based on the total weight of the nutritional oil.

[0344] Clause 212. The nutritional oil of any one of clauses 181 to 211, further comprising an added antioxidant.

[0345] Clause 213. The nutritional oil of clause 212, wherein the added antioxidant comprises a tocopherol.

[0346] Clause 214. The nutritional oil of any one of clauses 181 to 213, comprising 50 wt% or greater, 55 wt% or greater, 60 wt% or greater, 65 wt% or greater, 70 wt% or greater, 75 wt% or greater, 80 wt% or greater, or 85 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0347] Clause 215. The nutritional oil of any one of clauses 181 to 214, comprising 50 wt% or greater, preferably 55 wt% or greater, preferably 60 wt% or greater, preferably 65 wt% or greater, more preferably 70 wt% or greater, more preferably 75 wt% or greater, more preferably 80 wt% or greater, or more preferably 85 wt% or greater of triglycerides based on the total weight of the nutritional oil.

[0348] Clause 216. The nutritional oil of any one of clauses 181 to 215, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, less than or equal to 0.15 ppm, or less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0349] Clause 217. The nutritional oil of any one of clauses 181 to 216, comprising 3-MCPD in an amount less than or equal to 0.2 ppm, preferably less than or equal to 0.15 ppm, or more preferably less than or equal to 0.1 ppm based on the total amount of nutritional oil.

[0350] Clause 218. The nutritional oil of any one of clauses 181 to 217, comprising glycidol in an amount less than or equal to 0.1 ppm, less than or equal to 0.05 ppm, less than or equal to 0.04 ppm, less than or equal to 0.03 ppm, or less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0351] Clause 219. The nutritional oil of any one of clauses 181 to 218, comprising glycidol in an amount less than or equal to 0.1 ppm, preferably less than or equal to 0.05 ppm, preferably less than or equal to 0.04 ppm, more preferably less than or equal to 0.03 ppm, or more preferably less than or equal to 0.02 ppm based on the total amount of nutritional oil.

[0352] Clause 220. The nutritional oil of any one of clauses 181 to 219, wherein the nutritional oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

[0353] Clause 221. The nutritional oil of any one of clauses 181 to 220, wherein the nutritional oil has an anisidine value of less than 20, preferably less than 15, preferably less than 12, more preferably less than 10, or more preferably less than 8.

[0354] Clause 222. The nutritional oil of any one of clauses 181 to 221, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, less than or equal to 8 meq / kg, less than or equal to 5 meq / kg, less than or equal to 3 meq / kg, less than or equal to 2 meq / kg, or less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0355] Clause 223. The nutritional oil of any one of clauses 181 to 222, wherein the nutritional oil has a peroxide value of less than or equal to 10 meq / kg, preferably less than or equal to 8 meq / kg, preferably less than or equal to 5 meq / kg, preferably less than or equal to 3 meq / kg, more preferably less than or equal to 2 meq / kg, or more preferably less than or equal to 1 meq / kg based on the total weight of the nutritional oil.

[0356] Clause 224. The nutritional oil of any one of clauses 181 to 223, wherein the nutritional oil is a microbial oil.

[0357] Clause 225. The nutritional oil of clause 224, wherein the microbial oil is produced by Mortierella alpina.Experimental Examples

[0358] Having generally described the nutritional oils and associated methods, a further understanding can be obtained by reference to the following experimental examples provided herein. The following experimental examples are provided for purposes of illustration only and are not intended to be limiting.Example 1 - Comparative Example (Crude Oils)

[0359] Mortierella alpina was used to produce several samples of microbial oil containing greater than 400 mg / g arachidonic acid. The manufacturing equipment employed in the production of the microbial oil samples used mineral oil as a lubricant. The microbial oil samples were each extracted from biomass using hexane that had been recovered from a previous extraction process using a mineral oil-based system. Specifically, the hexane used to extract the present microbial oils had previously been used to extract other microbial oils from other biomass samples. The hexane had been separated from the previous microbial oils via evaporation and collected for reuse. Specifically, the effluent air leaving the solvent recovery system contained the evaporated hexane vapor and a mineral oil-based system was used toabsorb the residual hexane from the effluent air before the air was vented to the atmosphere. The mineral oil-based system contained a solid packing material to maximize the oil / vapor contact surface area. The adsorbed solvent was collected and reused to extract the present microbial oil samples.

[0360] The initial crude oil samples included greater than 0.1 wt% free fatty acids. Each of the crude oil samples were analyzed by GC-FID to determine the level of MOSH in the microbial oils. In further detail, a glass solid phase extraction (SPE) cartridge with PTFE frit was packed with a lower layer of activated aluminum oxide (1.5g) and an upper layer of silica gel impregnated with 10% silver nitrate (2.5g). The cartridge was conditioned by rinsing with several volumes of high purity hexane. Approximately 1.5 grams of the individual crude oil samples were respectively dissolved and quantitatively transferred to the cartridge with two 1 mL aliquots of hexane. The MOSH fraction of the crude oil samples was eluted with 5 mL of hexane added dropwise. An internal standard (n-octadecane) was added to the collected MOSH fraction. The samples were then concentrated to a final volume of approximately 1 mL under a gentle stream of nitrogen.

[0361] The concentrated sample was analyzed for MOSH by gas chromatography with flame ionization detection using an AGILENT 7890 GC-FID fitted with a “multi-mode” inlet upgrade.

[0362] Chromatographic Conditions:Injection ParametersInjection volume: 50pLInjection speed: 80pL / min.Inlet SettingsCarrier gas: HydrogenInlet mode: PTV Solvent VentInlet Pressure: 6.3143 psiSeptum purge flow: 3 mL / min.Split vent purge flow: 60 mL / min. after 4.5 min.Solvent vent purge flow: 100 mL / min. at 5 psi for 1 min.Inlet temperature: 45°C for 1 min., then ramp at 600°C / min. to 370°C. and hold.Oven SettingsCarrier gas flow: 2 mL / min. H2 (constant flow mode)Temperature: 50°C for 4 min., then ramp at 30°C / min. to 370°C, hold 5 min.Run Time: 19.7 min.Flame Ionization Detector ParametersTemperature: 370°CHydrogen flow: 40 mL / min.Air flow: 450 mL / min.Make-up flow (N2): 20 mL / min. (column + make-up)The inlet was held at close to the boiling point of the sample solvent (hexane) for 1 min. and the sample was injected slowly while applying a flow of carrier gas to sweep the evaporated solvent out of the purge vent, leaving any MOSH deposited on the inlet liner wall. After injection was complete, the inlet temperature was rapidly increased, and the flow was directed to the GC column to vaporize the analytes and transfer them to the column. The oven temperature was then increased to elute any MOSH as a broad band.

[0363] Quantification was achieved by comparing the total peak area of the MOSH peak to that of the internal standard peak. A system suitability standard was also injected to verify the performance of the chromatographic system. The limit of quantification for this method was determined to be 7 ppm. The results are depicted in Table 1 below:Table 1 - Mosh Levels in Comparative Samples (Crude Oils)Example 2 - Replace Mineral Oil-Based System (Crude Oils)

[0364] Mortierella alpina was used to produce several samples of microbial oil containing greater than 400 mg / g arachidonic acid. The manufacturing equipment employed in the production of the microbial oil samples used mineral oil as a lubricant. The microbial oil samples were each extracted from biomass using hexane that had been recovered from a previous extraction process using a sunflower oil-based system. Specifically, the hexane used to extract the present microbial oils had previously been used to extract other microbial oils from other biomass samples. The hexane had been separated from the previous microbial oils via evaporation and collected for reuse. Specifically, the effluent air leaving the solvent recovery system contained the evaporated hexane vapor and a sunflower oil-based system was used to absorb the residual hexane from the effluent air before the air was vented to the atmosphere. The sunflower oil-based system contained a solid packing material to maximize the oil / vapor contact surface area. The adsorbed solvent was collected and reused to extract the present microbial oil samples.

[0365] The initial crude oil samples included greater than 0.1 wt% free fatty acids. Each of the crude oil samples were analyzed by GC-FID to determine the level of MOSH in the microbial oils using the same methodology described in Example 1. The results are depicted in Table 2 below:Table 2 - MOSH Levels in Inventive Examples (Crude Oils)Example 3 - Stability of Alternative Solvents for Mineral Oil Scrubber System

[0366] The present example was directed to modeling a mineral oil-based scrubber system used to remove fugitive hexane after separation from extracted microbial oil. Mineral oil-based scrubber systems are typically part of the overall hexane solvent recovery unit and can reduce volatile organic compound (VOC) emissions. However, mineral oil is also a potential source of MOH, including MOSH and MOAH, contamination in the microbial oil when the captured hexane is recycled for further use in microbial oil extraction. As such, the present investigation looked at alternative recovery solvents to mineral oil that might be suitable for use in the scrubber system.

[0367] Three experiments were performed to mimic the mineral oil scrubber system, but replacing mineral oil with the following non-limiting examples: 1) high oleic sunflower oil (HOSO); 2) medium chain triglycerides (MCT); or 3) canola oil. In each of the experiments, 300 g of the alternative recovery solvent was introduced into a Parr 4500 series reactor.Reconstituted steam condensate was added to the system at 2.3 wt% and hexane was added at 1.4 wt%. The mixture was sealed and heated to 105 °C while mixing and maintaining pressure targeted at 20 psi. The reaction was sampled twice per day for a period of 5-9 days.

[0368] Each sample of the mixture was analyzed for free fatty acids (FFA) as an indicator of hydrolysis over time. As can be seen in FIG. 1 , the FFA content increases sharply in the HOSO sample after approximately 4 days, but only very gradually in MCT or canola during the period tested. HOSO has the highest amount of monounsaturated fatty acids, followed by canola and MCT, which is the most unsaturated. This data suggests that HOSO breaks down faster than canola and MCT and would need to be replaced more frequently in the scrubber system than canola and MCT. FFA transferred from recaptured hexane to subsequent batches of extracted microbial oil can generally be removed from the microbial oil more easily than MOSH and MOAH contamination from mineral oil.Example 4 - Adsorptive Capacity of Alternative Recovery Solvents

[0369] Oil was pumped to the top of a glass column using a Milton Roy volumetric piston pump. The oil was allowed to flow downward on the wall of the column by gravity and was collected into a glass flask. The oil flow rate was targeted to be a set volumetric rate. A flask ofn-hexane was heated and evaporated with entry into the bottom of the column. A sweep gas (nitrogen) was added to prevent condensation of the hexane, and to mimic a non-condensable load into the column as would typically be present in a production mineral oil scrubber system. The nitrogen sweep flow was measured qualitatively. The moisture (volatiles) was measured by infrared analysis to define the amount of solvent collected.

[0370] As can be seen in Table 3, Sample 1 and Sample 2 adsorbed n-hexane at pilot scale ranged from 10-14%, with HOSO slightly out-performing mineral oil adsorption. These trials were repeated with Samples 3 and 4 with reduced nitrogen sweep gas, and recovery was increased slightly to 11-22%. In both trials, HOSO recovered slightly more n-hexane solvent than mineral oil.Table 3 - Initial Adsorptive Capacity of HOSO vs. Mineral Oil

[0371] In Table 4, the nitrogen flowrate was reduced to as low as practical at lab scale, and mineral oil was compared to HOSO, MCT oil, and canola oil. Overall solvent adsorption increased as compared to the experiments in Table 3. The recovery was the highest using MCT oil, and the lowest using mineral oil. The HOSO and canola oil demonstrated similar n-hexane adsorption.Table 4 - Adsorptive Capacity of Recovery SolventsExample 5 - Replace Mineral Oil-Based System (Refined Oils)

[0372] Antioxidant blends are typically added to the final oils, which can include tocopherols. In some cases, tocopherols can also include some MOSH, and therefore contribute to the totalMOSH in the final oil. To evaluate the contributions of the tocopherols in the final oils, two separate tocopherol samples were sourced from different suppliers and analyzed to determine the MOSH contents of the samples. The MOSH content of the respective samples was determined using the following methodology.

[0373] Approximately 0.5 grams of tocopherols were accurately weighted into glass test tubes. 3 mL of reagent alcohol (denatured ethanol) and 1 ml_ of 45% potassium hydroxide (aq.) were added. The samples were capped and placed on a heating block set to 100°C for 30 minutes to saponify the lipid components. Non-saponifiable material was extracted with GC-MS grade hexanes. An internal standard (n-octadecane) was added for quantification purposes. The hexane layer was washed with deionized water and concentrated under a stream of nitrogen.

[0374] An empty glass solid phase extraction (SPE) cartridge with a PTFE frit was packed with a lower layer of 1 .5 grams of activated aluminum oxide and an upper layer of 2.5 grams of silica gel (impregnated with 10% silver nitrate). The cartridge was conditioned by rinsing with 20 mL of GC-MS grade hexane. The concentrated hexane extract from the sample was loaded onto the conditioned column. The MOSH fraction was then eluted with 5 mL of hexane. This hexane layer was concentrated to a volume of approximately 1 mL under a gentle stream of nitrogen.

[0375] The concentrated fraction was analyzed for MOSH content using an AGILENT 8890 Gas Chromatograph with a flame ionization detector (GC-FID). The instrument was fitted with a “multi-mode inlet” upgrade to allow for large volume injections for improved sensitivity.

[0376] A blank oil sample was prepared following the same steps, but omitting the tocopherol sample, to account for any minor MOSH contributions from reagents.

[0377] The inlet is held at close to the boiling point of the sample solvent (hexane) for 1 min. and the sample is injected slowly while applying a flow of carrier gas to sweep the evaporated solvent out of the purge vent, leaving any MOSH deposited on the inlet liner wall. After injection is complete, the inlet temperature is rapidly increased, and the flow is directed to the GC column to vaporize the analytes and transfer them to the column. The oven temperature is then increased to elute any MOSH as a broad band.

[0378] Chromatographic conditions:Injection ParametersInjection volume: 50pLInjection speed: 80pL / min.Inlet SettingsCarrier gas: HydrogenInlet mode: PTV Solvent VentInlet Pressure: 6.3143 psiSeptum purge flow: 3 mL / min.Split vent purge flow: 60 mL / min. after 4.5 min.Solvent vent purge flow: 100 mL / min. at 5 psi for 1 min.Inlet temperature: 45°C for 1 min., then ramp at 600°C / min. to 370°C. and hold.Oven SettingsCarrier gas flow: 2 mL / min. H2(constant flow mode)Temperature: 50°C for 4 min., then ramp at 30°C / min. to 370°C, hold 5 min.Run Time: 19.7 min.Flame Ionization Detector ParametersTemperature: 370°CHydrogen flow: 40 mL / min.Air flow: 450 mL / min.Make-up flow (N2): 20 mL / min. (column + make-up)The inlet is held at close to the boiling point of the sample solvent (hexane) for 1 min. and the sample is injected slowly while applying a flow of carrier gas to sweep the evaporated solvent out of the purge vent, leaving any MOSH deposited on the inlet liner wall. After injection is complete, the inlet temperature is rapidly increased, and the flow is directed to the GC column to vaporize the analytes and transfer them to the column. The oven temperature is then increased to elute any MOSH as a broad band.

[0379] Quantification is achieved by comparing the total peak area of the MOSH peak to that of the internal standard peak. A system suitability standard is also injected to verify the performance of the chromatographic system. The limit of quantification for this method was determined to be 2 ppm.

[0380] The results of the analysis of the two separate tocopherol samples is reported in Table 5 below.Table 5 - MOSH Content in Tocopherol Samples

[0381] As can be seen from Table 5, samples from one tocopherol supplier have considerably higher MOSH values than those of the other supplier.

[0382] Mortierella alpina was used to produce a crude microbial oil containing greater than 300 mg / g arachidonic acid in the same manner as described in Example 2. The initial crude oil samples included greater than 0.1 wt% free fatty acids. The crude oil samples were further refined using degumming, caustic refining, bleaching, winterization, and deodorization (RBWD) processes as are commonly used in the art.

[0383] An antioxidant blend was added to the refined oils to help protect against oxidative degradation of the oils. Specifically, two oil samples were prepared with the same level of tocopherols, but where Lot V7 from Table 5 was added to one of the two oil samples and Lot K4 from Table 5 was added to the other sample. These two refined oil samples were then analyzed to determine MOSH content using the same methodology described above for the tocopherol samples. The other data points were determined using appropriate respective methods as described elsewhere herein. The various characteristics of the oil samples are reported below in Table 6.Table 6 - Characteristics of Inventive Examples (Final Oils)Example 6 - ARA and DHA Blends

[0384] DHA crude oils were prepared from a Schizochytrium strain and extracted using aqueous extraction technology, as described in US 10,385,289 B2, which is incorporated herein by reference. The crude oil samples included greater than 550 mg / g DHA. The crude oil was further refined using degumming, caustic refining, bleaching, winterization, and deodorization processes commonly used in the oil refining industry. An antioxidant blend was added to the DHA final oil including tocopherol Lot K4 from Table 5. This DHA final oil was then blended at two different ratios (ARA: DHA) with the ARA final oil tested as Sample 2 in Table 6. The MOSH content for these two blends was analyzed using the same methodology as described in Example 5. The results are depicted in Table 7 below.Table 7 - MOSH content in ARA / DHA blends

Claims

1. Dietary oil containing: arachidonic acid (ARA) in an amount of 300 mg / g or more based on the total weight of dietary oil; free fatty acids in an amount of 0.1% or more by weight based on the total weight of the dietary oil; and less than 7 ppm mineral oil saturated hydrocarbons (MOSH) based on the total weight of dietary oil.

2. The dietary oil of claim 1, containing ARA in an amount of 350 mg / g or more, 400 mg / g or more, 450 mg / g or more, 500 mg / g or more, or 550 mg / g or more, based on the total weight of the dietary oil.

3. A dietary oil according to any one of the preceding claims, containing ARA in an amount of from 300 mg / g to 400 mg / g, from 350 mg / g to 450 mg / g, from 400 mg / g to 500 mg / g, from 450 mg / g to 550 mg / g, or from 500 mg / g to 600 mg / g, based on the total weight of the dietary oil.

4. A dietary oil according to any one of the preceding claims, containing MOSH in an amount of 6 ppm or less, 5 ppm or less, 4 ppm or less, or 3.5 ppm or less, based on the total weight of the dietary oil.

5. A dietary oil according to any one of the preceding claims, containing MOSH in an amount of from 0.5 ppm to less than 7 ppm or from 2 ppm to less than 7 ppm, based on the total weight of the dietary oil.

6. A dietary oil according to any one of the preceding claims, containing mineral oil aromatic hydrocarbons (MOAH) in an amount of 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil.

7. A dietary oil according to any of the preceding claims, further containing an added antioxidant.

8. The dietary oil of claim 7, wherein the added antioxidant comprises tocopherol.

9. A dietary oil according to any one of the preceding claims, containing 50% by weight or more, 55% by weight or more, 60% by weight or more, 65% by weight or more, 70% by weight or more, 75% by weight or more, 80% by weight or more, or 85% by weight or more triglycerides, based on the total weight of the dietary oil.

10. A dietary oil according to any one of the preceding claims, containing 3MCPD in an amount of 0.2 ppm or less, 0.15 ppm or less, or 0.1 ppm or less, based on the total weight of the dietary oil.

11. A dietary oil according to any one of the preceding claims, containing glycidol in an amount of 0.1 ppm or less, 0.05 ppm or less, 0.04 ppm or less, 0.03 ppm or less, or 0.02 ppm or less, based on the total weight of the dietary oil.

12. A dietary oil according to any one of the preceding claims, wherein the dietary oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

13. The dietary oil of any one of the preceding claims, wherein the dietary oil has a peroxide value of 10 meq / kg or less, 8 meq / kg or less, 5 meq / kg or less, 3 meq / kg or less, 2 meq / kg or less, or 1 meq / kg or less, based on the total weight of the dietary oil.

14. A dietary oil according to any one of the preceding claims, wherein the dietary oil is a microbial oil.

15. The dietary oil of claim 14, wherein the microbial oil is produced by Mortierella alpina.

16. Dietary oil containing: ARA in an amount of 300 mg / g or more based on the total weight of dietary oil; less than 0.1% by weight of free fatty acids based on the total weight of the dietary oil; and 3 ppm or less MOSH based on the total weight of dietary oil.

17. The dietary oil of claim 16, containing ARA in an amount of 350 mg / g or more, 400 mg / g or more, 450 mg / g or more, or 500 mg / g or more, based on the total weight of the dietary oil.

18. Dietary oil according to any one of paragraphs 16, 17, containing ARA in an amount of from 300 mg / g to 400 mg / g, from 350 mg / g to 450 mg / g, or from 400 mg / g to 500 mg / g, based on the total weight of the dietary oil.

19. The dietary oil of any one of paragraphs 16-18, containing MOSH in an amount of 2.5 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil.

20. A dietary oil according to any one of claims 16-18, containing MOSH in an amount of from 0.1 ppm to 2 ppm or from 0.5 ppm to 3 ppm based on the total weight of the dietary oil.

21. The dietary oil of any one of paragraphs 16-20, containing MOAH in an amount of 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil.

22. A dietary oil according to any one of paragraphs 16-21, further containing an added antioxidant.

23. The dietary oil of claim 22, wherein the added antioxidant comprises tocopherol.

24. The dietary oil of any one of claims 16 to 23, containing 60 wt.% or more, 65 wt.% or more, 70 wt.% or more, 75 wt.% or more, 80 wt.% or more, 85 wt.% or more, 90 wt.% or more, or 95 wt.% or more triglycerides, based on the total weight of the dietary oil.

25. The dietary oil of any one of claims 16 to 24, containing 3-MCPD in an amount of 0.2 ppm or less, 0.15 ppm or less, or 0.1 ppm or less, based on the total weight of the dietary oil.

26. The dietary oil of any one of paragraphs 16-25, containing glycidol in an amount of 0.1 ppm or less, 0.05 ppm or less, 0.04 ppm or less, 0.03 ppm or less, or 0.02 ppm or less, based on the total weight of the dietary oil.

27. The dietary oil of any one of claims 16-26, wherein the dietary oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

28. The dietary oil of any one of paragraphs 16-27, wherein the dietary oil has a peroxide value of 10 meq / kg or less, 8 meq / kg or less, 5 meq / kg or less, 3 meq / kg or less, 2 meq / kg or less, or 1 meq / kg or less, based on the total weight of the dietary oil.

29. The dietary oil of any one of claims 16 to 28, wherein the dietary oil is a microbial oil.

30. The dietary oil of claim 29, wherein the microbial oil is produced by Mortierella alpina.

31. A blend of dietary oils containing: a first dietary oil containing ARA mixed with a second dietary oil containing docosahexaenoic acid (DHA), wherein the mixture of dietary oils contains ARA and DHA in a weight ratio of ARA to DHA of from 0.5 to 3, and wherein the mixture of dietary oils contains MOSH in an amount of 4 ppm or less.

32. A dietary oil blend according to claim 31, containing ARA in an amount of 100 mg / g or more, 150 mg / g or more, 200 mg / g or more, 250 mg / g or more, 300 mg / g or more, or 350 mg / g or more, based on the total weight of the dietary oil blend.

33. A mixture of dietary oils according to any one of paragraphs 31, 32, containing ARA in an amount of from 100 mg / g to 200 mg / g, from 150 mg / g to 250 mg / g, from 200 mg / g to 300 mg / g, from 250 mg / g to 350 mg / g, or from 300 mg / g to 400 mg / g, based on the total weight of the mixture of dietary oils.

34. A dietary oil blend according to any one of paragraphs 31-33, containing DHA in an amount of 100 mg / g or more, 150 mg / g or more, 200 mg / g or more, 250 mg / g or more, 300 mg / g or more, or 350 mg / g or more, based on the total weight of the dietary oil blend.

35. A dietary oil mixture according to any one of paragraphs 31-34, containing DHA in an amount of from 100 mg / g to 200 mg / g, from 150 mg / g to 250 mg / g, from 200 mg / g to 300 mg / g, from 250 mg / g to 350 mg / g, or from 300 mg / g to 400 mg / g, based on the total weight of the dietary oil mixture.

36. A mixture of dietary oils according to any one of paragraphs 31-35, containing ARA and DHA in a weight ratio of ARA to DHA of from 0.5 to 1.5, from 1 to 2, from 1.5 to 2.5, or from 2 to 3.

37. A dietary oil blend according to any one of paragraphs 31-36, containing MOSH in an amount of 3.5 ppm or less, 3 ppm or less, 2.5 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil blend.

38. A dietary oil blend according to any one of paragraphs 31-36, containing MOSH in an amount of from 0.1 ppm to 2 ppm, from 0.5 ppm to 3 ppm, from 1 ppm to 4 ppm, or from 1.5 ppm to 5 ppm, based on the total weight of the dietary oil blend.

39. A dietary oil blend according to any one of paragraphs 31-38, containing MOAH in an amount of 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil blend.

40. A mixture of dietary oils according to any one of paragraphs 31-38, further containing an added antioxidant.

41. A mixture of dietary oils according to claim 40, wherein the added antioxidant comprises tocopherol.

42. A dietary oil mixture according to any one of paragraphs 31-41, containing 60 wt.% or more, 65 wt.% or more, 70 wt.% or more, 75 wt.% or more, 80 wt.% or more, 85 wt.% or more, 90 wt.% or more, or 95 wt.% or more triglycerides, based on the total weight of the dietary oil mixture.

43. A dietary oil blend according to any one of paragraphs 31-42, containing 3-MCPD in an amount of 0.2 ppm or less, 0.15 ppm or less, or 0.1 ppm or less, based on the total weight of the dietary oil blend.

44. A dietary oil blend according to any one of paragraphs 31-43, containing glycidol in an amount of 0.1 ppm or less, 0.05 ppm or less, 0.04 ppm or less, 0.03 ppm or less, or 0.02 ppm or less, based on the total weight of the dietary oil blend.

45. A blend of dietary oils according to any one of paragraphs 31-44, wherein the blend of dietary oils has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

46. ​​A dietary oil blend according to any one of paragraphs 31-45, wherein the dietary oil blend has a peroxide value of 10 meq / kg or less, 8 meq / kg or less, 5 meq / kg or less, 3 meq / kg or less, 2 meq / kg or less, or 1 meq / kg or less, based on the total weight of the dietary oil blend.

47. A mixture of dietary oils according to any one of paragraphs 31-46, wherein the mixture of dietary oils comprises microbial oil.

48. A blend of dietary oils according to claim 47, wherein the microbial oil contains DHA.

49. A dietary oil mixture according to claim 48, wherein the microbial oil is produced by Crypthecodinium cohnii.

50. A mixture of dietary oils according to claim 48, wherein the microbial oil is produced by a microorganism of the genus Thraustochytrium or Schizochytrium.

51. A blend of dietary oils according to claim 47, wherein the microbial oil comprises ARA.

52. A mixture of dietary oils according to claim 51, wherein the microbial oil is produced by Mortierella alpina.

53. A dietary oil blend according to any one of claims 47-52, comprising a first microbial oil containing ARA and a second microbial oil containing DHA.

54. A method for producing dietary oil according to any one of paragraphs 1-30, comprising: i. combining the LC-PUFA containing oil with an organic solvent; ii. removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude dietary oil; and iii. recovering at least a portion of the organic solvent removed in step ii using a recovery solvent, wherein the recovery solvent contains less than 50% by weight of mineral oil based on the total weight of the recovery solvent, and dietary oil contains less than 7 ppm MOSH.

55. A method for producing dietary oil, comprising: i. combining the LC-PUFA containing oil with an organic solvent; ii. removing at least a portion of the organic solvent from the LC-PUFA-containing oil to obtain a crude dietary oil; and iii. recovering at least a portion of the organic solvent removed in step ii using a recovery solvent, wherein the recovery solvent contains less than 50% by weight of mineral oil based on the total weight of the recovery solvent.

56. The method according to any one of paragraphs 54, 55, wherein the organic solvent comprises hexane.

57. The method of any one of paragraphs 54-57, wherein the organic solvent comprises at least a portion of recovered organic solvent previously combined with the second LC-PUFA-containing oil.

58. The method of claim 58, wherein the recovered organic solvent contains less than 100 ppm MOSH, less than 75 ppm MOSH, less than 50 ppm MOSH, less than 25 ppm MOSH, less than 10 ppm MOSH, less than 5 ppm MOSH, or less than 1 ppm MOSH.

59. The method of any one of claims 54-58, wherein combining the LCPUFA-containing oil with the organic solvent comprises extracting the microbial oil from the microorganism using the organic solvent.

60. The method according to claim 59, wherein the microorganism belongs to the order Mortierellales.

61. The method according to claim 60, wherein the microorganism is Mortierella alpina.

62. The method according to claim 59, wherein the microorganism belongs to the order Thraustochytriales.

63. The method according to claim 62, wherein the microorganism belongs to the genus Thraustochytrium or Schizochytrium.

64. The method according to claim 59, wherein the microorganism is a dinoflagellate.

65. The method of claim 64, wherein the microorganism is Crypthecodinium cohnii.

66. The method according to any one of paragraphs 54-65, in which the recovery solvent contains less than 40 wt.%, less than 30 wt.%, less than 20 wt.%, less than 10 wt.%, or less than 5 wt.% mineral oil, based on the total weight of the recovery solvent.

67. The method according to any one of paragraphs 54-66, wherein the solvent for recovery comprises triglycerides.

68. The method of any one of paragraphs 54-67, wherein the recovery solvent comprises sunflower oil, medium chain triglycerides, canola oil, or a combination thereof.

69. The method according to any one of paragraphs 54-68, wherein the dietary oil is crude dietary oil.

70. The method of claim 69, wherein the dietary oil contains free fatty acids in an amount of more than 0.1 wt.%, more than 0.15 wt.%, more than 0.2 wt.%, more than 0.25 wt.%, more than 0.3 wt.%, more than 0.35 wt.%, more than 0.4 wt.%, more than 0.45 wt.%, more than 0.5 wt.%, or more than 0.55 wt.%, based on the total weight of the dietary oil.

71. The method according to any one of paragraphs 54-68, in which the dietary oil is a finished dietary oil obtained by further processing crude dietary oil to obtain a finished dietary oil.

72. The method according to any one of paragraphs 54-71, in which the dietary oil contains free fatty acids in an amount of less than 0.1 wt.% or less than 0.05 wt.% based on the total weight of the dietary oil.

73. The method of any one of paragraphs 54-72, wherein the dietary oil contains one or more of ARA, DHA or EPA in an amount of 300 mg / g or more based on the total weight of the dietary oil.

74. The method of any one of paragraphs 54-73, wherein the dietary oil comprises MOSH in an amount of 6 ppm or less, 5 ppm or less, 4 ppm or less, 3 ppm or less, 2.5 ppm or less, 2 ppm or less, 1.5 ppm or less, 1 ppm or less, or 0.5 ppm or less, based on the total weight of the dietary oil.

75. The method of any one of paragraphs 54-73, wherein the dietary oil contains MOSH in an amount of from 0.1 ppm to 2 ppm, from 0.5 ppm to 3 ppm, from 1 ppm to 4 ppm, or from 1.5 ppm to 5 ppm, based on the total weight of the dietary oil.

76. The method of any one of paragraphs 54-75, wherein the dietary oil contains MOAH in an amount of 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil.

77. The method of any one of paragraphs 54-76, further comprising using equipment material containing 50% by weight or less mineral oil based on the total weight of the equipment material.

78. The method of claim 77, wherein the equipment material is an equipment lubricant.

79. The method of any one of claims 77 and 78, wherein the equipment material comprises canola oil, sunflower oil, mixed chain triglycerides, or a combination thereof.

80. Dietary oil obtained by the method according to any of paragraphs 54-79, containing: ARA in an amount of 300 mg / g or more based on the total weight of dietary oil; and from 0.5 ppm to 10 ppm MOSH based on the total weight of dietary oil.

81. The dietary oil of claim 80, containing ARA in an amount of 350 mg / g or more, 400 mg / g or more, 450 mg / g or more, 500 mg / g or more, or 550 mg / g or more, based on the total weight of the dietary oil.

82. Dietary oil according to any one of paragraphs 80, 81, containing ARA in an amount of from 300 mg / g to 400 mg / g, from 350 mg / g to 450 mg / g, from 400 mg / g to 500 mg / g, from 450 mg / g to 550 mg / g, or from 500 mg / g to 600 mg / g, based on the total weight of the dietary oil.

83. A dietary oil according to any one of paragraphs 80-82, containing free fatty acids in an amount of 0.1% by weight or more based on the total weight of the dietary oil.

84. The dietary oil of claim 83, containing MOSH in an amount of from 0.5 ppm to 2 ppm, from 1 ppm to 3 ppm, from 2 ppm to 4 ppm, from 3 ppm to 5 ppm, from 4 ppm to 6 ppm, or from 5 ppm to 7 ppm, based on the total weight of the dietary oil.

85. A dietary oil according to any one of paragraphs 80-82, containing free fatty acids in an amount of less than 0.1% by weight based on the total weight of the dietary oil.

86. The dietary oil of claim 85, containing MOSH in an amount of from 0.1 ppm to 2 ppm, from 0.5 ppm to 3 ppm, from 1 ppm to 4 ppm, or from 1.5 ppm to 5 ppm, based on the total weight of the dietary oil.

87. The dietary oil of any one of paragraphs 80-86, containing MOAH in an amount of 5 ppm or less, 4 ppm or less, 3 ppm or less, 2 ppm or less, 1.5 ppm or less, or 1 ppm or less, based on the total weight of the dietary oil.

88. A dietary oil according to any one of paragraphs 80-87, further comprising an added antioxidant.

89. The dietary oil of claim 85, wherein the added antioxidant comprises tocopherol.

90. A dietary oil according to any one of paragraphs 80-89, containing 50% by weight or more, 55% by weight or more, 60% by weight or more, 65% by weight or more, 70% by weight or more, 75% by weight or more, 80% by weight or more, or 85% by weight or more triglycerides, based on the total weight of the dietary oil.

91. The dietary oil of any one of paragraphs 80-90, containing 3-MCPD in an amount of 0.2 ppm or less, 0.15 ppm or less, or 0.1 ppm or less, based on the total weight of the dietary oil.

92. The dietary oil of any one of paragraphs 80-91, containing glycidol in an amount of 0.1 ppm or less, 0.05 ppm or less, 0.04 ppm or less, 0.03 ppm or less, or 0.02 ppm or less, based on the total weight of the dietary oil.

93. The dietary oil of any one of paragraphs 80-92, wherein the dietary oil has an anisidine value of less than 20, less than 15, less than 12, less than 10, or less than 8.

94. The dietary oil of any one of paragraphs 80-93, wherein the dietary oil has a peroxide value of 10 meq / kg or less, 8 meq / kg or less, 5 meq / kg or less, 3 meq / kg or less, 2 meq / kg or less, or 1 meq / kg or less, based on the total weight of the dietary oil.

95. A dietary oil according to any one of paragraphs 80-94, wherein the dietary oil is a microbial oil.

96. Dietary oil according to claim 95, wherein the microbial oil is produced by Mortierella alpina.