CN113747796A

CN113747796A - Process for making monoacylglycerol oils and food products containing monoacylglycerol oils

Info

Publication number: CN113747796A
Application number: CN202080018774.0A
Authority: CN
Inventors: D·J·蒙迪希罗; W·J·布斯曼
Original assignee: Grecos Biological Co ltd
Current assignee: Grecos Biological Co ltd; Glycosbio Inc
Priority date: 2019-01-18
Filing date: 2020-01-17
Publication date: 2021-12-03
Also published as: AU2020207945A1; CA3127093A1; WO2020150661A9; EP3911164A1; WO2020150661A1; JP2022517271A

Abstract

Compositions and methods for incorporating processing oils with high Monoacylglyceride (MAG) content into products and food products are provided. In particular, methods of producing high MAG content process oils are provided.

Description

Process for making monoacylglycerol oils and food products containing monoacylglycerol oils

Cross Reference to Related Applications

The present application claims priority from U.S. provisional application No. 62/794,412 filed on day 18, 2019 and U.S. provisional application No. 62/833,558 filed on day 12, 2019, each of which is incorporated herein by reference in its entirety.

Background

A chronic defect in pancreatic digestive enzyme secretion is known as Exocrine Pancreatic Insufficiency (EPI). Without these digestive enzymes, patients with EPI cannot properly digest the nutritional components of the food and may develop malnutrition and abdominal disturbances. EPI is common in individuals with chronic pancreatitis and several other chronic gastrointestinal disorders. EPI is also found in patients with cystic fibrosis. Pancreatic Enzyme Replacement Therapy (PERT), in which an individual administers an enzyme capsule each time the individual consumes food, can reduce the effects of EPI. Conventionally, PERT therapy involves pancreatin extracted from porcine pancreas.

Lipids are energy-intensive compounds and are a source of essential long-chain fatty acids. The consumed lipids, which typically include a high percentage of Triacylglycerides (TAG), are digested with pancreatic secreted lipases to form Free Fatty Acids (FFA) and Monoacylglycerides (MAG). Preventing the release of lipases from the pancreas results in very poor digestion of triacylglycerol-containing fats and oils. For patients with EPI, this may lead to severe malnutrition, as calories, essential fatty acids and fat-soluble nutrients will be trapped in the undigested lipid particles and pass through the system.

The clinical need for alternative sources of nutrition is not met and individuals with EPI need not be supplemented with PERT to consume the alternative sources of nutrition.

Partially hydrolyzed fats and oils in the form of MAG are readily absorbed by individuals with EPI without PERT. MAG oil-based products have been clinically evaluated as capsule-based nutritional supplements; however, capsules are used to avoid bad taste. For conventional MAG oil sources, the starting oil is chemically or enzymatically treated to make MAG, which is then extracted and distilled with a solvent to fractionate MAG from the other components of the starting oil. These MAG products are sold as relatively pure products, containing only negligible contaminating Free Fatty Acids (FFA), (diacylglycerides) DAG and TAG, and few other compounds. Thus, conventional sources of MAG oil are often deficient in other natural compounds found in the oil, such as tocopherols.

There is a clinical need for very high caloric density nutritional products that can be consumed by individuals with inefficient or impaired digestive systems. In addition to individuals with pancreatic pathologies (e.g., cystic fibrosis, pancreatitis, and pancreatic cancer patients), other patients diagnosed with or not diagnosed with Exocrine Pancreatic Insufficiency (EPI) would also benefit from the product. Furthermore, individuals with bile dysfunction (cholestasis) may benefit from "predigesting" fat that does not require bile acids for emulsification. There are high calorie "energy bars" and beverages on the market. However, these products are not suitable for individuals who cannot digest (hydrolyze) the fat in the product. To date, no one has formulated lipids into liquid (milkshakes) and solid (sticks) forms suitable for "PERT-free" use. These preparations can serve as a source of "complete nutrition" to supply all of the individual's caloric and essential fatty acid needs. The liquid nutrition may be in the form of an Oral Nutritional Supplement (ONS) product or an enteral feeding product.

Thus, there is a need for high caloric density foods that can be consumed by individuals with inefficient or impaired digestive systems. The present application describes a method of producing an edible enzyme-modified oil (EMO) substantially free of TAG.

Monochloropropanediol (MCPD) and glycidyl esters are formed during the refining of edible oils. These compounds are toxic to humans and need to be minimized in food products. According to the international agency for research on cancer (IARC), 3-MCPD is currently classified as a possible human carcinogen (group 2B). IARC and the national toxicology program of the united states classify glycidol as potentially carcinogenic to humans (group 2A). Current standards recommend exposure to less than 2ug/kg body weight per day, less than 140 ug/day for a 70kg body weight person, and only 10 ug/day for a 5kg (about 10lb) infant.

MCPD compounds were first detected in acid (HCl) hydrolyzed proteins, but MCPD esters were found to be present in refined vegetable oils in 2008. This problem has been found to be common. Many edible oils are processed to remove components that negatively impact appearance, taste, shelf stability, safety, and consumer acceptance. Monoacyl and diacylglycerol (MAG and DAG) in the oil can react with chloride ions in the deodorization process to produce 3-monochloropropanediol (3-MCPD) ester and Glycidyl Ester (GE).

Table 1 below shows the levels of MCPD compounds in US samples of edible oil (from Food Additives and Contaminants Part A (Part A), vol.30, vol.2013, 2081, 2092, 12).

The estimated exposure of us infants to 3-MCPD and glycidyl ester consumption of infant formula was recently reported (j.spungen et al, part a of food additives and pollutants, vol 35, No. 6, 1085-. In this analysis, U.S. FDA data on 3-MCPD and glycidyl ester concentrations (3-MCPD and glycidyl equivalent, respectively) in a small convenient sample of infant formula were used to estimate exposure through consumption of formula by infants between 0 and 6 months of age. Based on the average concentration in all formulations, the 3-MCPD and GE exposures were estimated to be 7-10 μ g/kg bw/day and 2 μ g/kg bw/day, respectively. The average exposure estimated by consuming a single manufacturer's produced formulation ranged from 1 μ g/kg bw/day to 14 μ g/kg bw/day for 3-MCPD and from 1 μ g/kg to 3 μ g/kg for GE.

Accordingly, there is a need for methods of reducing and eliminating these compounds in edible oils. The present method of producing edible Enzyme Modified Oils (EMOs) substantially free of TAGs and contaminants such as MCPD thus meets this need.

Some vegetable oils are rich in oleic acid. These oils have proven beneficial for health. Some of these reasons may be attributed to oleic acid and oils rich in such fatty acids, such as olive oil, almond oil and canola (rapeseed) oil. Triolein is a TAG rich in oleic acid, as shown below. Triolein is oleic acid 100% esterified to glycerol at all three positions. These three positions are defined as "sn-1, sn-2 and sn-3". Sn is a "stereo number".

In view of the health benefits of these oils, there is a need for compositions and methods that can provide the health benefits of these oils to individuals with EPI, and enhance and make the beneficial components of the oils more readily available.

Disclosure of Invention

The present disclosure relates to a product comprising a process oil derived from an oil source. In one embodiment, the processing oil comprises a MAG content equal to or greater than 30 wt% of the total weight of the processing oil, a DAG content of about 10 wt% to about 30 wt% of the total weight of the processing oil, and a FFA content of about 5 wt% to about 60 wt% of the total weight of the processing oil, wherein the processing oil is TAG free or comprises a TAG content equal to or less than 5 wt% of the total weight of the processing oil, and wherein the processing oil comprises non-oil components derived from and naturally present in the oil source such that the non-oil components are not added to the processing oil.

In some embodiments, the oil source is from a source selected from a plant, an animal, a fish, or a mixture thereof. In some embodiments, the oil source comprises an MCPD compound. In these examples, the process oil is substantially free of MCPD compounds.

In some embodiments, the non-oil ingredients of the product are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In some embodiments, the product comprises greater than 1% MAG by weight of the total weight of the product.

In some embodiments, the product comprises greater than 50% MAG by weight of the total weight of the product.

The present disclosure also relates to a food product. In one embodiment, the food product comprises oil and has a caloric density of about 1 kcal/gram to about 5 kcal/gram, wherein about 20% to about 50% of the calories originate from the oil.

In some embodiments, the oil of the food product is a process oil derived from an oil source, wherein process oil comprises a MAG content equal to or greater than 30 wt% of the total weight of process oil, a DAG content of about 10 wt% to about 30 wt% of the total weight of process oil, and a FFA content of about 5 wt% to about 60 wt% of the total weight of process oil, wherein the process oil is free of TAG or comprises a TAG content equal to or less than about 5 wt% of the total weight of process oil, and wherein the process oil comprises non-oil ingredients derived from an oil source and naturally present in an oil source such that the non-oil ingredients are not added to the process oil.

In some embodiments, the oil source of the food product is from a source selected from a plant, an animal, or a fish.

In some embodiments, the non-oil ingredients of the food product are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In some embodiments, the food product comprises greater than 1% MAG by weight of the total weight of the product.

In some embodiments, the food product comprises greater than 50% MAG by weight of the total weight of the product.

In some embodiments, the food product has a total weight of about 25 grams to about 3000 grams.

In some embodiments, the food product has a total caloric content of from about 1kcal to about 5kcal per gram.

In some embodiments, the food product may further comprise a source of carbohydrates.

In some embodiments, the food product may further comprise a protein source.

In some embodiments, the oil comprises 5% to 95% of the total caloric content of the food product.

The present disclosure also relates to a method of making a monoacylglycerol-rich oil. In one embodiment, the method comprises mixing a starting oil comprising Triacylglycerols (TAG), a buffer solution, and a first enzyme capable of hydrolyzing the TAG to Free Fatty Acids (FFA) to produce a first reaction mixture; reacting the reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAG for a first period of time to produce an aqueous phase and a lipid (free fatty acid) reaction product; inactivating the first enzyme in the reaction product; collecting the lipid reaction product; mixing the lipid reaction product with food grade glycerol and a second enzyme capable of esterifying FFA to form a second reaction mixture; reacting the second reaction mixture for a second period of time to produce a reaction product lipid oil phase and a glycerin phase; inactivating the second enzyme in the reaction product; adding a salt to the reaction and separating a lipid oil phase from the glycerol phase; and collecting the lipid oil phase.

In some embodiments, the starting oil is an oil derived from a plant, an animal, a marine organism, or a mixture thereof. In some embodiments, the starting oil comprises an MCPD compound, and the lipid oil phase is substantially free of the MCPD compound.

In some embodiments, the first enzyme is lipase AY.

In some embodiments, the first time period is a time period sufficient to hydrolyze at least 94% of the TAGs in the starting oil.

In some embodiments, the first period of time is between about 14 hours and 24 hours.

In some embodiments, the step of reacting the reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAG is performed at a temperature between about 30 ℃ to about 35 ℃.

In some embodiments, the steps of mixing a starting oil comprising a Triacylglycerol (TAG), a buffer solution, and a first enzyme capable of hydrolyzing the TAG to a Free Fatty Acid (FFA) and reacting the reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAG to a FFA are performed under a nitrogen atmosphere.

In some embodiments, the second enzyme is lipase G.

In some embodiments, the second time period is a time period sufficient to result in an enrichment of MAG in the lipid oil phase of about 60% to 95%.

In some embodiments, the second time period is between about 24 hours and about 72 hours.

In some embodiments, the step of reacting the second reaction mixture for a second period of time to produce a lipid oil phase and a glycerol phase is conducted at a temperature between about 17 ℃ and 23 ℃.

In some embodiments, the method further comprises drying the reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the reaction product.

In some embodiments, said step of drying said reaction product is performed at a temperature between 20 ℃ and 30 ℃.

In some embodiments, the drying step is applied throughout the second time period.

In some embodiments, the step of inactivating the second enzyme is performed by heating the reaction product.

In some embodiments, the heating is performed at a temperature of at least 70 ℃ for at least 1 hour.

In some embodiments, the step of separating the lipid oil phase from the glycerol phase comprises adding sodium chloride to the reaction product.

In some embodiments, the final concentration of sodium chloride comprises up to 0.3 wt% sodium chloride.

In some embodiments, the method further comprises reestablishing a nitrogen atmosphere over the lipid reaction product prior to mixing the lipid reaction product with food-grade glycerol and a second enzyme capable of esterifying FFA and glycerol.

In some embodiments, said steps of removing at least a portion of said aqueous phase and replacing said at least a portion of said aqueous phase with an approximately equivalent volume of water and waiting for a second period of time are repeated prior to performing said step of collecting said lipid reaction product.

In some embodiments, the method further comprises adding tocopherol to the lipid oil phase after collecting the lipid oil phase.

In some embodiments, the process oil having an overall fatty acid content comprises oleic acid Monoglyceride (MOG) in an amount of between about 5% to about 75% by weight of the overall fatty acid content of the process oil composition.

In some embodiments, the processing oil comprises oleic acid and linoleic acid in a ratio of between about 0.01 to about 5, the processing oil having greater than 50% by weight Monoacylglycerides (MAG), based on the total weight of the processing oil.

In some embodiments, the processing oil comprises oleic acid and linolenic acid in a ratio of between about 1 to about 100, the processing oil having greater than 50% by weight Monoacylglycerides (MAG), based on the total weight of the processing oil.

In some embodiments, the process oil comprises oleic acid and linoleic acid, and has an overall fatty acid content, wherein linoleic acid is present in an amount from about 10% to about 90% by weight of the overall fatty acid content of the process oil.

In some embodiments, the fatty acid profile of the process oil is substantially the same as the fatty acid profile of a pre-process oil from which the process oil is produced.

In some embodiments, the process oil has a fatty acid profile comprising oleic acid, linoleic acid, and linolenic acid, wherein the amounts of oleic acid, linoleic acid, and linolenic acid are within about 10% of the amounts of oleic acid, linoleic acid, and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In some embodiments, the process oil has a fatty acid profile comprising oleic acid, linoleic acid, and linolenic acid, wherein the amounts of oleic acid, linoleic acid, and linolenic acid are within about 1% of the amounts of oleic acid, linoleic acid, and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In some embodiments, a method for promoting glucose homeostasis in a subject in need thereof comprises the step of administering to the subject a composition comprising a processing oil comprising oleic acid monoglyceride, wherein at least 50% by weight of the oleic acid monoglyceride is a 1-oleyl monoglyceride.

In some embodiments, a method for treating type II diabetes in a subject in need thereof comprises the step of administering to the subject a composition comprising a processed oil comprising monoglycerides of oleic acid, wherein at least 50% by weight of the monoglycerides of oleic acid are 1-oleyl monoglycerides.

In some embodiments, a method for promoting glucose homeostasis in a subject in need thereof comprises the step of administering to the subject a composition comprising a processed oil of the present disclosure.

In some embodiments, a method for treating diabetes in a subject in need thereof comprises the step of administering to the subject a composition comprising a processed oil of the present disclosure.

Drawings

Figure 1 depicts TLC separation of components of starting vegetable oil, intermediate FFA and final MAG oil.

Figure 2 depicts TLC separation of components of starting vegetable oil, intermediate FFA and final MAG oil.

Figure 3 depicts the distribution of FFA, MAG, DAG and TAG in "Ensure original nutritional milkshake" and the enzyme-modified oil products of the present disclosure ("GBFS").

Fig. 4 depicts a block flow diagram of a process for making an enzyme-modified oil.

Fig. 5 depicts the distribution of amino acids and peptides in the ready-to-drink nutritional beverage and GBFS hydrolyzed pea protein.

Fig. 6A depicts the NMR spectrum of authentic TAG (glyceryl tristearate).

Fig. 6B depicts NMR spectra of enzyme modified oils produced from almond oil.

Fig. 7 depicts the rise in serum triglycerides following ingestion of an EMO-based ready-to-drink milkshake.

Figure 8 depicts the elevation of serum triglycerides following ingestion of the fert-free MAG-based RTDS.

Figure 9 depicts blood glucose levels (mean) of patients after intake of MAG-based RTDS without PERT or standard nutritional supplement with PERT.

Figure 10 depicts serum triglyceride levels (mean) of patients after intake of MAG-based RTDS without PERT or standard nutritional supplement with PERT.

Fig. 11 depicts the percentage of certain fatty acids in the canola oil and canola EMO of example 13 and two commercial monoglyceride samples.

Fig. 12A depicts the amounts of various compounds in canola oil and canola EMO of example 14 based on LC/MS analysis.

Fig. 12B depicts the amounts of various compounds in canola oil and canola EMO of example 14 based on LC/MS analysis.

Detailed Description

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. In this application, the use of the singular includes the plural, the words "a" or "an" mean "at least one" and the use of "or" means "and/or" unless specifically stated otherwise. Furthermore, the use of the term "including" and other forms such as "including" and "included" is not limiting. Furthermore, terms such as "element" or "component" encompass an element or component comprising one unit as well as an element or component comprising more than one unit unless specifically stated otherwise.

The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. All documents, or portions of documents, cited in this application, including but not limited to patents, patent applications, articles, books, and treatises, are hereby expressly incorporated by reference in their entirety for any purpose. In the event that one or more of the incorporated documents and similar materials define a term that conflicts with the definition of that term in the present application, the present application controls.

"enriched" in the context of the present invention means that the amount thereof is higher than in the starting material. For example, a MAG-rich oil is an oil having a MAG content greater than the initial MAG content prior to the enrichment process, or the starting oil has a higher percentage of MAG than the oil processed prior to the enrichment process. The enrichment process can be performed by converting the TAG to MAG, thereby increasing MAG content and percentage and decreasing TAG content or percentage.

Triacylglycerols ("TAGs"), also known as triglycerides, are glycerides consisting of three fatty acid chains covalently bonded to a glycerol molecule through ester chain bonds. TAGs can also be classified as having a long or medium chain length. Long chain TAGs contain fatty acids having 14 or more carbons, while medium chain TAGs contain fatty acids having 6 to 12 carbons. The long chain TAGs may comprise omega-3 and omega-6 fatty acids. The medium chain TAGs have saturated fatty acids, and thus do not contain omega-3 or omega-6 fatty acids. Long chain tag (lct) and Medium Chain Triglycerides (MCT) can be used as energy sources.

Diacylglycerols ("DAGs"), also known as diglycerides, are glycerides consisting of two fatty acid chains covalently bonded to the glycerol molecule through ester chain bonds.

Monoacylglycerols ("MAG"), also known as monoglycerides, are glycerides consisting of one fatty acid chain covalently bonded to a glycerol molecule through an ester chain bond.

As used herein, the term "processing oil" refers to a non-naturally occurring oil composition that is substantially free of, or has a reduced amount of, TAG relative to a pre-modified or pre-processed oil.

As used herein, the term "enzyme-modified oil" or "EMO" refers to a processing oil in which TAG is enzymatically converted to MAG, such as, for example, using the enzymatic conversions of the present disclosure.

As used herein, the term "food product" refers to a manufactured or non-naturally occurring food product. It should be understood that the food products referred to herein, while manufactured as a whole and not naturally occurring, may include various combinations of natural ingredients, where the combinations are not naturally occurring, or where the combinations are indeed naturally occurring, which are not present in the relative amounts used in the food product.

The terms "patient," "individual," and "subject" are used interchangeably herein and refer to a mammalian subject, preferably a human patient, to be treated. In some cases, the methods of the invention are used in laboratory animals, veterinary applications, and development of animal models of disease, including but not limited to rodents, including mice, rats and hamsters, and primates.

"treatment" refers to an intervention designed to prevent the development of a disorder or to alter the pathology or symptomatology of a disorder. Thus, "treatment" may refer to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those already suffering from the disorder as well as those in need of prevention of the disorder. In the treatment of tumors (e.g., cancers), a therapeutic agent may directly reduce the pathology of tumor cells, or render tumor cells more amenable to treatment by other therapeutic agents, such as radiation and/or chemotherapy.

As used herein, a "non-oil component" is a component that naturally occurs in an oil source that is not MAG, DAG, TAG, FFA, or lipid.

In some embodiments, the starting oil may include, for example, but is not limited to, a vegetable-derived oil, such as olive oil, almond oil, canola oil, coconut oil, cottonseed oil, palm kernel oil, palm olein oil, palm stearin oil, peanut oil, linseed oil, sunflower seed oil, corn oil, grape seed oil, palm oil, soybean oil, or an animal-derived oil, such as fish oil, sardine oil, or anchovy oil, or algae oil, or a mixture of any of the foregoing vegetable and/or animal oils. In one aspect, the starting oil comprises a mixture of olive oil, sunflower seed oil, and linseed oil, wherein about 50% to about 80% by weight of the total weight of the starting oil is olive oil, about 10% to about 30% by weight of the total weight of the starting oil is sunflower seed oil, and about 5% to about 20% by weight of the total weight of the starting oil is linseed oil. In another aspect, about 50% to about 80% by weight of the total weight of the starting oil is olive oil, about 10% to about 30% by weight of the total weight of the starting oil is linseed oil, and about 5% to about 20% by weight of the total weight of the starting oil is sunflower seed oil.

In some embodiments, the starting oil comprises an MCPD compound. The amount of the MCPD compound may be from about 0.30mg/kg to about 12.0mg/kg, or from about 1.0mg/kg to about 11.00mg/kg, or from about 2.00mg/kg to about 10.50 mg/kg. Examples of starting oils that include MCPD compounds include almond, canola, coconut, corn, cottonseed, grape seed, olive, palm kernel, palm olein, palm stearin, peanut, soybean, and sunflower.

In some embodiments, the process of making a MAG-rich product includes a first step of hydrolyzing the TAG. For example, but not limited thereto, hydrolysis of TAG may be performed by a lipase such as lipase AY (Amano Enzymes, USA Elgin IL, USA), or any non-regiospecific lipase which cleaves at sn-1, sn-2 and sn-3 positions.

In some embodiments, the first step of hydrolyzing the TAG can be performed at a temperature of about 30 ℃ to 35 ℃. For example, but not limited to, the first step of hydrolyzing TAG can be performed at a temperature of 30 ℃ to 35 ℃, 31 ℃ to 35 ℃, 32 ℃ to 35 ℃, 33 ℃ to 35 ℃, 34 ℃ to 35 ℃, 30 ℃ to 34 ℃, 31 ℃ to 34 ℃, 32 ℃ to 34 ℃, 33 ℃ to 34 ℃, 30 ℃ to 33 ℃, 31 ℃ to 33 ℃, 32 ℃ to 33 ℃, 30 ℃ to 32 ℃, 31 ℃ to 32 ℃, 30 ℃ to 31 ℃ or 30 ℃, 31 ℃, 32 ℃, 33 ℃, 34 ℃ or 35 ℃.

In some embodiments, the first step of hydrolyzing the TAG can be performed for about 14 hours to 24 hours. For example, but not limited to, the first step of hydrolyzing TAG may be performed for 14 hours to 20 hours, 14 hours to 16 hours, 18 hours to 24 hours, 22 hours to 24 hours, 18 hours to 20 hours, or about 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, or 24 hours.

In some embodiments, the first step of hydrolyzing the TAG results in hydrolysis of substantially all of the TAG. For example, but not limited to, the first step of hydrolyzing the TAG results in hydrolysis of 94% to 100%, 95% to 100%, 96% to 100%, 97% to 100%, 98% to 100%, 99% to 100%, 94% to 99%, 95% to 99%, 96% to 99%, 97% to 99%, 98% to 99%, 94% to 98%, 95% to 98%, 96% to 98%, 97% to 98%, 94% to 97%, 95% to 97%, 96% to 97%, 94% to 96%, 95% to 96%, or 94% to 95% of the TAG or at least 94%, 95%, 96%, 97%, 98%, 99%, or 100% of the TAG.

In some embodiments, the process of making a MAG-rich product includes a second step of esterification with glycerol to enrich the MAG oil content. For example, but not limited thereto, this second esterification step may be performed by a lipase such as lipase G (Amano Enzymes, USA Elgin IL, USA) or any region-specific lipase which catalyses the esterification of the sn-1 position but which does not effectively catalyse the formation of second or third esters on glycerol (to make DAG and TAG).

In some embodiments, the second step of esterification with glycerol to enrich for MAG oil content results in a MAG enrichment in the product of about 70% to 95%. For example, but not limited to, MAG oil content may be enriched by 70% to 95%, 75% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 70% to 90%, 75% to 90%, 80% to 90%, 85% to 90%, 70% to 85%, 75% to 85%, 80% to 85%, 70% to 80%, 75% to 80%, 70% to 75% or 70%, 75%, 80%, 85%, 90% or 95%.

In some embodiments, the second step of esterification with glycerol may be performed at a temperature of about 17 ℃ to 23 ℃. For example, but not limited to, esterification with glycerol can be performed at a temperature of 17 ℃ to 23 ℃, 18 ℃ to 23 ℃, 19 ℃ to 23 ℃, 20 ℃ to 23 ℃, 21 ℃ to 23 ℃, 22 ℃ to 23 ℃, 17 ℃ to 22 ℃, 18 ℃ to 22 ℃, 19 ℃ to 22 ℃, 20 ℃ to 22 ℃, 21 ℃ to 22 ℃, 17 ℃ to 21 ℃, 18 ℃ to 21 ℃, 19 ℃ to 21 ℃, 20 ℃ to 21 ℃, 17 ℃ to 20 ℃, 18 ℃ to 20 ℃, 19 ℃ to 20 ℃, 17 ℃ to 19 ℃, 18 ℃ to 18 ℃ or 17 ℃, 18 ℃, 19 ℃, 20 ℃, 21 ℃, 22 ℃ or 23 ℃.

In some embodiments, the second step of esterification with glycerol may be performed for about 24 hours to 72 hours. For example, but not limited to, the second step of esterification with glycerol may be performed for 24 hours to 72 hours, 36 hours to 72 hours, 48 hours to 72 hours, 60 hours to 72 hours, 24 hours to 60 hours, 36 hours to 60 hours, 48 hours to 60 hours, 24 hours to 48 hours, 36 hours to 48 hours, 24 hours to 36 hours or 24 hours, 30 hours, 36 hours, 42 hours, 48 hours, 54 hours, 60 hours, 66 hours, or 72 hours.

In some embodiments, the process of making a MAG-rich product includes a third step of inactivating the lipase and phase separating.

The resulting products from the above examples resulted in processing oils having a MAG content equal to or greater than 40 wt.%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 40 wt% to about 99 wt%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 50 wt% to about 99 wt%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 60 wt% to about 99 wt%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 70 wt% to about 99 wt%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 80 wt% to about 99 wt%, based on the total weight of the processing oil. In certain aspects, the MAG content is about 50 wt% to about 80 wt%, based on the total weight of the processing oil. In any of the above aspects, the TAG content is equal to or less than 5 wt% based on the total weight of the processing oil. In any of the above aspects, the TAG content is equal to or less than 4 wt%, equal to or less than 3 wt%, equal to or less than 2 wt%, equal to or less than 1 wt%, based on the total weight of the processing oil.

The resulting products from the above examples typically result in a process oil that is substantially free of MCPD compounds, but also when the starting oil has greater than 0.30mg/kg of MCPD compounds and in some cases from about 1.0mg/kg to about 12.00 mg/kg. More specifically, the process oil obtained by the above method comprises less than 100mg/kg of MCPD and even reaches levels undetectable by standard assays. As used herein, the term "substantially free" with respect to the amount of MCPD levels in the process oil of the present invention means levels below the detection limit of the assay described in example 11 below.

The resulting product from the above example yielded a process oil rich in MAG, in particular 1-MAG (MAG esterified at the sn-1 position). In the case of process oils made from starting oils having an oleic acid content, this results in the production of 1-oleyl monoacylglycerides (1-OG). It has been found that the fabrication process embodiments of the present disclosure can efficiently convert even sn-2 oleic acid TAGs to 1-OG. For example, starting from triolein, the manufacturing process embodiments of the present disclosure can produce three 1-OG molecules from a single triolein molecule, whereas digestion of triolein by pancreatic lipase can only produce one third of the total oleic acid in the form of monoacylglycerols, particularly as 2-monoacylglycerols (2-OG) rather than 1-OG, since lipases release the oleic acid moieties at the sn-1 and sn-3 positions as free fatty acids. Thus, the processing oil produced by the manufacturing method of the present disclosure can produce up to three times the 1-OG content as compared to the normal digestion process. The reaction scheme below shows the conversion of triolein to 2-OG and two free oleic acid molecules (process a-normal digestion) and the conversion of triolein to 3 1-OG molecules by the process of the present disclosure (process B).

It has also been found that the manufacturing process embodiments of the present disclosure can maintain the fatty acid profile of the starting oil in the process oil. Table 2 below shows representative fatty acid profiles for various starting oils, which table provides the percentage of total Fatty Acid Methyl Esters (FAMEs). Nd-is uncertain. SAF-safflower oil, GRP-grape oil, SIL-silybum marianum oil, HMP-sesame oil, SFL-sunflower oil, WHG-wheat germ oil, PMS-pumpkin seed oil, SES-sesame oil, RB-rice bran oil, ALM-almond oil, RPS-rapeseed (canola) oil, PNT-peanut oil, OL-olive oil and COC-coconut oil. Table 2 also provides the oleic acid/linoleic acid and oleic acid/linolenic acid ratios for each oil.

In particular, linoleic (omega-3 fatty acids) and linolenic (omega-6 fatty acids) acids are known to be essential to humans because they can be used to produce longer and more desaturated fatty acids, also known as long chain polyunsaturated fatty acids (LC-PUFAs), comprising eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA).

In some embodiments, a product comprises a process oil of the present disclosure. In some embodiments, the process oil itself is a separate product.

In some embodiments, the processing oil comprises a MAG content equal to or greater than 30 wt% of the total weight of the processing oil. For example, but not limited to, the process oil includes a MAG content of about 30 to 95 wt.%, 40 to 95 wt.%, 50 to 95 wt.%, 60 to 95 wt.%, 70 to 95 wt.%, 80 to 95 wt.%, 90 to 95 wt.%, 30 to 90 wt.%, 40 to 90 wt.%, 50 to 90 wt.%, 60 to 90 wt.%, 70 to 90 wt.%, 80 to 90 wt.%, 30 to 80 wt.%, 40 to 80 wt.%, 50 to 80 wt.%, 60 to 80 wt.%, 70 to 80 wt.%, 30 to 70 wt.%, 40 to 70 wt.%, 50 to 70 wt.%, 60 to 70 wt.%, 30 to 60 wt.%, 40 to 60 wt.%, or 60 to 95 wt.% of the total weight of the process oil, 50 to 60, 30 to 50, 40 to 50, or about 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95 wt%.

In some embodiments, including the above embodiments with respect to MAG content, the processing oil comprises a DAG content equal to or greater than 5 wt% of the total weight of the processing oil. For example, but not limited to, the process oil includes a MAG content of about 5 wt.% to 66 wt.%, 10 wt.% to 66 wt.%, 20 wt.% to 66 wt.%, 30 wt.% to 66 wt.%, 40 wt.% to 66 wt.%, 50 to 66 wt.%, 5 wt.% to 50 wt.%, 10 wt.% to 50 wt.%, 20 wt.% to 50 wt.%, 30 wt.% to 50 wt.%, 40 wt.% to 50 wt.%, 5 wt.% to 40 wt.%, 10 wt.% to 40 wt.%, 20 wt.% to 40 wt.%, 30 wt.% to 40 wt.%, 5 wt.% to 30 wt.%, 10 wt.% to 30 wt.%, 20 wt.% to 30 wt.%, 5 wt.% to 20 wt.%, 10 wt.% to 20 wt.%, 5 wt.% to 10 wt.%, or about 5 wt.%, 10 wt.%, 15 wt.%, 20 wt.%, 25 wt.%, based on the total weight of the process oil, 30, 35, 40, 45, 50, 55, 60, or 66 wt%.

In some embodiments, including the embodiments described above with respect to MAG and/or DAG content, the processing oil comprises an FFA content equal to or greater than 5 wt% of the total weight of the processing oil. For example, but not limited to, the processing oil includes MAG in an amount of about 5 to 66 wt.%, 10 to 66 wt.%, 20 to 66 wt.%, 30 to 66 wt.%, 40 to 66 wt.%, 50 to 66 wt.%, 5 to 50 wt.%, 10 to 50 wt.%, 20 to 50 wt.%, 30 to 50 wt.%, 40 to 50 wt.%, 5 to 40 wt.%, 10 to 40 wt.%, 20 to 40 wt.%, 30 to 40 wt.%, 5 to 30 wt.%, 10 to 30 wt.%, 20 to 30 wt.%, 5 to 20 wt.%, 10 to 20 wt.%, 5 to 15 wt.%, 10 to 15 wt.%, 5 to 10 wt.%, or about 5 wt.%, based on the total weight of the processing oil, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or 66 wt%.

In some embodiments, including the above embodiments with respect to MAG, DAG, and/or FFA content, the processing oil is free of TAG or comprises a TAG content equal to or less than 5 wt% of the total weight of the processing oil. For example, but not limited to, the processing oil includes a TAG content of about 0 wt% to 5 wt%, 1 wt% to 5 wt%, 2 wt% to 5 wt%, 3 wt% to 5 wt%, 4 wt% to 5 wt%, 0 wt% to 4 wt%, 1 wt% to 4 wt%, 2 wt% to 4 wt%, 3 wt% to 4 wt%, 0 wt% to 3 wt%, 1 wt% to 3 wt%, 2 wt% to 3 wt%, 0 wt% to 2 wt%, 1 wt% to 2 wt%, 0 wt% to 1 wt%, or 0 wt%, 1 wt%, 2 wt%, 3 wt%, 4 wt%, or 5 wt% of the total weight of the processing oil. By way of further example, but not limitation, TAG content may be less than 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1%.

In some embodiments, the processing oil comprises about 30 wt% to 80 wt% MAG, about 10 wt% to about 30 wt% DAG, about 0 wt% to about 5 wt% TAG, and about 5 wt% to about 60 wt% FFA, based on the total weight of the processing oil.

In any embodiment herein, the processing oil comprises non-oil components derived from and naturally occurring in the oil source such that the non-oil components are not added to the processing oil. For example, but not limited to, such non-oil ingredients may comprise antioxidants such as tocopherols, which comprise alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol or gamma-tocotrienol, as well as other vitamins such as vitamin K and structurally similar 2-methyl-1, 4-naphthoquinone (3-) derivatives. In some embodiments, the antioxidant is selected from natural (e.g., mixed tocopherols or ascorbic acid) and synthetic (e.g., butylated hydroxyanisole or butylated hydroxytoluene) antioxidants. Further examples of non-oil ingredients include ceramide phosphate, monogalactosyldiacylglycerol, phosphatidylmethanol, sitosterol ester, campesterol ester, sphingolipids, phosphatidylglycerol, wax esters, and sphingomyelin.

In any of the embodiments herein, the process oil comprises Monoglycerol Oleate (MOG) and has a total fatty acid content. MOG can comprise from about 5% to about 75% by weight of the total fatty acid content of the processed oil. In some embodiments, for example and without limitation, MOG can comprise from about 10 wt% to about 75 wt%, from about 20 wt% to about 75 wt%, from about 30 wt% to about 75 wt%, from about 40 wt% to about 75 wt%, from about 50 wt% to about 75 wt%, or from about 60 wt% to about 75 wt% of the total fatty acid content of the processed oil. In some embodiments, the processed oil comprises oleic acid in an amount from about 5% to about 75% by weight of the total fatty acid content of the processed oil. In some embodiments, for example, but not limited to, oleic acid may comprise from about 10% to about 75%, from about 20% to about 75%, from about 30% to about 75%, from about 40% to about 75%, from about 50% to about 75%, or from about 60% to about 75% by weight of the total fatty acid content of the processed oil.

In any of the embodiments herein, the processing oil comprises MOG, and the MOG comprises at least 50 wt% of 1-oleyl monoglyceride (1-OG) of the total amount of MOG. For example, but not limiting of, the processing oil can include at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of the total MOG of 1-OG.

In any embodiment herein, the process oil comprises oleic acid in an amount from about 10% to about 75% by weight of the total fatty acid content of the process oil. For example, but not limited to, oleic acid may be present in an amount of from about 10% to about 75%, from about 20% to about 75%, from about 30% to about 75%, from about 40% to about 75%, from about 50% to about 75%, from about 60% to about 75%, from about 10% to about 20%, from about 10% to about 30%, from about 10% to about 40%, from about 10% to about 50%, or from about 10% to about 60% by weight of the total fatty acid content of the processed oil. In any embodiment herein, the oleic acid is esterified at the sn-1 position. For example, but not limited to, the process oil can include at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% of the total amount of oleic acid esterified at the sn-1 position.

In any embodiment herein, the processed oil comprises linoleic acid in an amount from about 1.5% to about 90% by weight of the total fatty acid content of the processed oil. For example, but not limited to, linoleic acid may be present in an amount from about 10% to about 90%, from about 20% to about 90%, from about 30% to about 90%, from about 40% to about 90%, from about 50% to about 90%, from about 60% to about 90%, from about 70% to about 90%, from about 80% to about 90%, from about 10% to about 25%, from about 10% to about 20% by weight of the total fatty acid content of the processed oil. In some embodiments, the processing oil comprises oleic acid and linoleic acid, and the ratio of oleic acid to linoleic acid is between about 0.01 and 5. For example, but not limited to, the ratio of oleic acid to linoleic acid can be between about 1 to about 5, about 1 to about 4, about 1 to about 3, about 1 to about 2, about 1.5 to about 5, about 1.5 to about 4, about 1.5 to about 3, about 1.5 to about 2, about 2 to about 5, about 2 to about 4, about 2 to about 3, about 2.5 to about 5, about 2.5 to about 4.5, about 2.5 to about 4, about 2.5 to about 3.5, about 2.5 to about 3, about 3 to about 5, or about 3 to about 4.

In any embodiment herein, the process oil comprises linolenic acid in an amount of about 0.01% to about 2% by weight of the total fatty acid content of the process oil. For example, but not limited to, linolenic acid may be present in an amount of from about 0.1% to about 2%, from about 0.5% to about 2%, from about 1% to about 2%, or from about 1.5% to about 2% by weight of the total fatty acid content of the processed oil. In some embodiments, the processing oil comprises oleic acid and linolenic acid, the ratio of oleic acid to linolenic acid being from about 1 to about 100. For example, but not limited to, the ratio of oleic acid to linolenic acid can be between about 10 to about 100, about 20 to about 100, about 30 to about 100, about 40 to about 100, about 50 to about 100, about 60 to about 100, about 70 to about 100, about 10 to about 90, about 10 to about 80, about 10 to about 70, about 10 to about 60, about 10 to about 50, about 10 to about 40, about 10 to about 30, or about 10 to about 20. It is to be understood that the reference to fatty acids in the claims is not to be construed as being limited to free fatty acids only and may refer to fatty acids in the form of glycerides. In the process oil of the present disclosure, the fatty acid will be present predominantly in the MAG form, whereas in the pre-process oil it will be present predominantly in the TAG form. In some cases, the fatty acids may to a lesser extent be in free form or DAG form. In process oils, the free fatty acids will generally be less than 10%.

In any of the embodiments herein, the fatty acid profile of the process oil is substantially the same as the fatty acid profile of a pre-process oil from which the process oil is produced. For example, but not limited to, the fatty acid profile of the processed oil can be within about 10%, about 9%, about 8%, about 7%, about 6%, about 5%, about 4%, about 3%, about 2%, or about 1% for each component of the fatty acid profile. In some embodiments, the fatty acid profile includes oleic acid, linoleic acid, and linoleic acid. However, any combination of fatty acids may form a fatty acid profile. For example, but not limiting of, the fatty acid profile can include any combination of the fatty acids listed in table 2 and EPA and DHA.

In any of the examples herein, the processing oil comprises three times the amount of MAG molecules compared to the number of TAG molecules in a pre-processing oil that produces the oil. For example, the amount of oleic acid in the form of MAG in the process oil may be up to three times the amount of oleic acid in the form of TAG in the pre-process oil on a molar basis. For example, but not limiting of, the amount of oleic acid in the form of MAG in the process oil may be 1.1, 1.25, 1.5, 1.75, 2.0, 2.25, 2.5, 2.75, or 3 times the amount of oleic acid in the form of TAG in the pre-process oil. For example, each oleic acid moiety in the form of a TAG in the pre-processed oil may be converted to MAG oleate, such that up to three oleic acid moieties, as in triolein, will be converted to MAG oleates. In some embodiments, the amount of MAG in the process oil is three times the amount of TAG in the pre-process oil from which the process oil is produced. For example, the amount of MAG produced corresponds to about three times the amount of TAG in the pre-processed oil. In some embodiments, the process oil comprises oleic acid in the form of oleic acid monoglyceride in an amount that is about the same as the amount of oleic acid in a pre-process oil from which the process oil is produced.

In some embodiments, the product or food product may include at least 1% MAG. For example, but not limited to, the food product may include at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% or more MAG by weight of the product or food product and any range or amount therebetween. As a further example, but not limiting of, the product or food product may comprise from 5 wt% to 15 wt% MAG based on the total weight of the product or food product.

In some embodiments, the product or food product may comprise at least 3 wt% of the processing oil of any embodiment herein, based on the total weight of the product or food product. For example, but not limited to, the food product can include at least 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or more of the processing oil of any embodiment herein and any range or amount therebetween by weight of the product or food product.

In some embodiments, the food product may further comprise a source of carbohydrates. For example, but not limited to, such carbohydrate sources may include monosaccharides, such as glucose and fructose, derived from carbohydrate sources, such as fruit and agave syrup. Other carbohydrate sources include other plant-based syrups, starches, and sugar alcohols.

In some embodiments, the food product may further comprise a protein source. In certain aspects, the protein source may be hydrolyzed or partially hydrolyzed. For example, but not limited to, such protein sources may include milk proteins (casein and whey), and other plant proteins, including proteins from soy, rice and rice bran, lentils, chickpeas, peanuts, almonds, spirulina (algae), quinoa, fungal proteins, chia seeds, and hemp seeds. Hydrolysed proteins can be extensively hydrolysed, with pea proteins being rich in peptides of 1 to 10 amino acids in length. In some embodiments, the protein is enriched in peptides of 1 to 10 amino acids in length by about 25% to 75% compared to commercial partially hydrolyzed proteins and other whey-based hydrolysates such as Peptamen and Crucial. For example, but not limited to, a protein is enriched in peptides of 1 to 10 amino acids in length by at least 25% to 75%, 35% to 75%, 45% to 75%, 55% to 75%, 65% to 75%, 25% to 65%, 35% to 65%, 45% to 65%, 55% to 65%, 25% to 55%, 35% to 55%, 45% to 55%, 25% to 45%, 35% to 45%, 25% to 35% or 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, or 75%.

In some embodiments, the food product may be a liquid, a semi-solid, or a solid. For example, but not limited to, the semi-solid may comprise a pudding, mousse, ice-lolly or ice cream-like product. For example, but not limiting of, the liquid may be a milkshake or other drink. For example, but not limited to, the solid may be a bar or other solid food product.

In some embodiments, the product or food product further comprises a viscosity altering agent. The viscosity altering agent may be, for example, but not limited to, xanthan gum or acacia gum. In some embodiments, the product or food product may further comprise structure or stability enhancing components such as, but not limited to, gum arabic, sunflower lecithin, and xanthan gum. In some embodiments, the product or food product further comprises a fiber source, such as, but not limited to, oligosaccharides. In some embodiments, the product or food product may further include a food preservative such as, but not limited to, sodium benzoate or potassium sorbate.

In some embodiments, the product or food product further comprises a flavoring agent, a masking agent, or a blocking agent. For example, but not limited to, the flavoring agent may comprise chocolate, vanilla, strawberry, or other flavoring agent.

In some embodiments, the food product has a total weight of at least 25 grams. For example, but not limited to, the food product may have a weight of at least 25 grams, 50 grams, 100 grams, 250 grams, 500 grams, 1000 grams, 1500 grams, 2000 grams, 2500 grams, 3000 grams, or more. In some embodiments, the food product has a total weight of about 25 grams to about 3000 grams. For example, but not limited to, the food product may have 25 to 3000 grams, 25 to 2500 grams, 25 to 2000 grams, 25 to 1000 grams, 25 to 500 grams, 50 to 3000 grams, 50 to 2500 grams, 50 to 2000 grams, 50 to 1500 grams, 50 to 1000 grams, 50 to 500 grams, 100 to 3000 grams, 100 to 2500 grams, 100 to 2000 grams, 100 to 1500 grams, 100 to 1000 grams, 100 to 500 grams, 250 to 3000 grams, 250 to 2500 grams, 250 to 2000 grams, 250 to 1500 grams, 250 to 1000 grams, 250 to 500 grams, 500 to 3000 grams, 500 to 2500 grams, 500 to 1500 grams, 500 to 1000 grams, 1000 to 3000 grams, 1000 to 2500 grams, 1000 to 2000 grams, 1000 to 1500 grams, 1500 to 3000 grams, 2000 to 3000 grams, 25 to 2000 grams, 25 to 250 grams, 100 to 250 grams, 250 to 2500 grams, 250 to 3000 grams, 250 grams, 100 grams to 500 grams, 100 grams to 500 grams, and 500 grams, 100 grams, and 500 grams, respectively, and 500 grams, respectively, and 500 grams, and 500 grams, and 500, respectively, and 500 grams, and 500, respectively, and 500, respectively, and 500, and, A weight of 25 grams to 50 grams, or a total weight of less than or equal to 25 grams, 50 grams, 100 grams, 150 grams, 200 grams, 250 grams, 300 grams, 350 grams, 400 grams, 450 grams, 500 grams, 1000 grams, 1500 grams, 2000 grams, 2500 grams, or 3000 grams.

In some embodiments, the food product has a total caloric content of about 200kcal to 1000 kcal. For example, but not limited to, the food product can have a caloric content of about 200kcal to 1000kcal, 400kcal to 1000kcal, 600kcal to 1000kcal, 800kcal to 1000kcal, 200kcal to 800kcal, 400kcal to 800kcal, 600kcal to 800kcal, 200kcal to 600kcal, 400kcal to 600kcal, 200kcal to 400kcal, or an amount less than or equal to 200kcal, 300kcal, 400kcal, 500kcal, 600kcal, 700kcal, 800kcal, 900kcal, or 1000 kcal.

In some embodiments, about 20% to 75% of the calories in the food product are derived from oil or fat. In one aspect, the oil or fat is a processed oil of any of the embodiments herein. In yet another aspect, the processing oil has a MAG content equal to or greater than 30 wt.% of the total weight of the processing oil. In other aspects, the MAG content of the processing oil is about 40 wt% to about 99 wt% of the total weight of the processing oil. In yet another aspect, the TAG content of the processing oil is less than 5 wt% of the total weight of the processing oil. For example, but not limited to, 20% to 50%, 30% to 50%, 40% to 50%, 20% to 40%, 30% to 40%, 20% to 30%, 20% to 75%, 30% to 75%, 40% to 75%, 50% to 75%, 60% to 75%, 70% to 75%, 20% to 70%, 30% to 70%, 40% to 70%, 50% to 70%, 60% to 70%, 20% to 60%, 30% to 60%, 40% to 60%, 50% to 60% or about 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, in a food product, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, or 75% of the heat is derived from any of the above-described process oils.

In any of the above embodiments with respect to a food product, about 20% to 50% of the calories in the food product are derived from a carbohydrate source. For example, but not limited to, 20% to 50%, 30% to 50%, 40% to 50%, 20% to 40%, 30% to 40%, 20% to 30% or about 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% of the calories in the food product are derived from a carbohydrate source.

In any of the above embodiments with respect to a food product, about 10% to 50% of the calories in the food product are derived from a protein source. For example, but not limited to, 10% to 50%, 20% to 50%, 30% to 50%, 40% to 50%, 10% to 40%, 20% to 40%, 30% to 40%, 10% to 30%, 20% to 30%, 10% to 20% or about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49% or 50% of the calories in the food product are derived from a protein source.

It should be understood that the above disclosed embodiments of the food product may be combined.

In some embodiments, the food products of the present disclosure comprise the processed oils of the present disclosure.

In some embodiments, the EMO-based product is administered by an individual with a dysfunction of the digestive system, such as, but not limited to, an individual with EPI or an individual taking PERT with food. In some embodiments, the EMO-based product is administered by an individual who desires to more quickly or completely convert lipids to serum triglycerides. Accordingly, a method for feeding a human or animal subject with a dysfunction of the digestive system is provided. The method comprises administering to the patient a food product as described in any one or a combination of the above embodiments. In one aspect, the human or animal subject has EPI. In other aspects, the human or animal subject has cystic fibrosis, pancreatitis, pancreatic cancer, or cholestasis.

In some embodiments, there is provided a method for promoting glucose homeostasis in a subject in need thereof, comprising administering to the subject a composition comprising a processed oil of the present disclosure. In some embodiments, the subject has a disorder that affects glucose homeostasis. In some embodiments, the disorder is insulin resistance or diabetes. In some embodiments, the disorder is type II diabetes.

In some embodiments, there is provided a method for treating diabetes in a subject in need thereof, comprising administering to the subject a composition comprising a processed oil of the present disclosure. In some embodiments, the diabetes is type II diabetes.

Examples of the invention

Example 1: a process for making a MAG-rich product compared to the starting TAG-rich oil.

The process of making a MAG-rich product comprises 3 key steps compared to the TAG-rich starting oil: (1) a mild enzyme-catalyzed reaction that hydrolyzes Triglycerides (TAGs) in a sequence that converts natural oils to a specific combination of FFAs, MAGs, DAGs and low residual TAGs; (2) esterification with glycerol, yielding a predominantly high level of MAG, leaving a low concentration of FFA; and (3) isolating the modified lipid product; this is achieved by phase separation with or without the aid of a centrifuge.

Step 1 enzymatic conversion of triacylglycerols

And (4) preparing a buffer solution. A sodium citrate solution (100mM, pH 5.8) was prepared in a stirred tank reactor. 11.1L of Deionized (DI) water was placed in the mixing vessel, the agitator was set at 200RPM, and 0.213kg of citric acid (anhydrous) was added. After the powder was dissolved, the pH was adjusted to 5.8 (about 0.121kg) with sodium hydroxide solution. 220mL were removed for subsequent enzyme preparations.

Preparation of enzyme solution. In a separate 250mL bottle, the enzyme solution was prepared under mild stirring: 200mL of citrate buffer solution was placed in the mixing vessel. 10g of AMANO lipase AY was added and the bottle was shaken until the enzyme was dissolved.

Conversion of triglyceride oil mixture. Three vegetable oils, olive oil, linseed oil and sunflower seed oil, were added to the container to obtain a total of 10kg of vegetable oil mixture. Vacuum was applied to reduce the pressure to about 20mmHg and the material was degassed (particularly from any dissolved oxygen). The stirring was set at 200RPM and the mixture was heated to 33 ℃ and stirred for about 15Min to remove any dissolved gases. The vacuum was then replaced with a nitrogen atmosphere. After the mixture was well dispersed, 220mL of the enzyme preparation in buffer was added. Stirring was continued and the reaction was monitored for 24 hours until complete conversion to FFA according to TLC analysis.

The reactor temperature was raised to 70 ℃ and stirring was continued for 1 hour to inactivate the enzyme.

Stirring was stopped and the phases were allowed to separate after about 60 minutes.

The aqueous phase (lower) and a small amount of the oil phase were removed to ensure removal of residual protein at the interface.

And 2, esterifying with glycerol. Typically, when FFAs are re-esterified with glycerol, they will produce a mixture of MAG, DAG and TAG. We have found that by significantly lowering the temperature (below 25 ℃) and removing the water formed in the reaction (by evaporation) the proportion of MAG in the product can be highly enriched (at least 60%, but up to 95%). This is unexpected.

In the reaction, the reaction product from step 1 (about 10L) was cooled to about 30 ℃ and stirred at 300 RPM. 10kg of food grade glycerol was added to the lipid mixture and the temperature was maintained at about 30 ℃. The mixture was stirred to produce a dispersion of oil and glycerol. To dry the reaction mixture, vacuum was applied: vacuum (25mmHg, torr) was first applied and the receiver was in place to collect the water. After evaporation of the residual water was stopped, 20G of Amano lipase G dissolved in water (50mL) was added to the reactor. The temperature was reduced to 23 ℃ using an oil-diaphragm pump and a cold trap to collect water, and the vacuum was changed to 5 mmHg. The mixture was stirred at 300RPM under vacuum at 23 ℃ for 72 hours, at which time the vacuum was broken and the mixture blanketed with nitrogen. The mixture was analyzed by TLC after 72 hours to assess conversion to MAG as shown in figure 1.

And 3, inactivating lipase and separating the lipase. After completion of the reaction, the lipase was inactivated by heating the mixture (under a blanket of nitrogen) to 70 ℃ for 1 hour. At this point, the MAG oil and glycerol are well mixed and difficult to separate by conventional gravity or centrifugation methods. After a number of experiments, we found that lipids can be separated from excess glycerol by adding 0.3 wt% of salt (NaCl) to the reaction mixture with stirring. The product mixture was then cooled to about 60 ℃ and allowed to stand without stirring for about 1 hour.

The lipid oil phase is separated from the remaining heavier glycerol phase. Removal of the glycerol phase: it contains some salts, residual water and dissolved inactive enzymes contained in the visible interface. The glycerol phase can be reused after membrane filtration and should be kept recycled. Tocopherol (vitamin E) was added to give a product oil with a concentration of 200ppm (0.02 wt%). Hydrolyzed oil (about 10kg is ready and can be stored under nitrogen blanket).

And (5) storing the product. The final hydrolysate was transferred to a nitrogen-covered food grade container for storage and transport.

Example 2: characterization of the oil produced in example 1.

The reaction products and the overall process can be evaluated using thin layer chromatography and gas chromatography.

And (4) testing by thin layer chromatography. The fractions of the oil samples were separated using TLC plates (Analtech Uniplate silica gel GHL, containing inorganic binders, 20x20cm, 250 μm). The solvent is hexane: diethyl ether: acetic acid (70:30:1) solution. Typical sample size is 3 μ L. After the solvent front approached the top of the plate (about 1cm), the plate was removed from the TLC tank and the solvent was evaporated in a fume hood. The components were visualized with iodine vapor (at room temperature) in a TLC pot and the relative intensities were estimated by colorimetric imaging (Amersham 600 imager). After 15 minutes in the jar, the plate was removed and photographed. After 30-60 minutes, the spot intensity decreased.

Physical property determinations were made to establish product consistency, color, water content, fats and oils (miscible).

Figure 1 depicts the final results of steps 1 to 3 in the process of making a MAG-rich product compared to the starting TAG-rich oil described in example 1. Figure 2 shows that the tocopherol originally present in olive oil was preserved according to the procedure described in example 1. In fig. 2, it can be seen that the tocopherol spots track over the TAG spots of all three lanes.

Fatty acid profile test-gas chromatography. Following derivatization to fatty acid methyl esters, the lipid components were analyzed and compared to standards, comprising C10:0 decanoic acid, C12:0 lauric acid, C14:0 myristic acid, C16:0 palmitic acid, C18:0 stearic acid, C18:1 oleic acid, C18:2 linoleic acid, and C18:3 alpha linolenic acid. For derivatization, a sample (500. mu.l) was added to a 5-ml reaction tube containing 2ml of boron trifluoride solution (12% in methanol), 20. mu.l of dimethoxypropane and 100. mu.l of tridecanoic acid internal standard solution (10 mg/ml). The reaction tube was vortexed and incubated in a60 ℃ heat block for 30 minutes.

The reaction tube was removed from the heating block and allowed to cool for 15 minutes. Then, 1ml of distilled water was added to quench the reaction, and then 1ml of hexane was added. The reaction tube was vortexed for 60 seconds and the phases were allowed to separate for 3 minutes. The top (hydrophobic) phase was transferred to a 1.5-ml tube containing about 50mg sodium sulfate (anhydrous). After vortexing for 60 seconds, the 1.5-ml tube was centrifuged and approximately 500. mu.l of the clear, dried hydrophobic phase was transferred to a gas chromatography sample vial.

Samples were analyzed using an Agilent 7890A gas chromatograph with a flame ionization detector and Agilent Openlab CDS Chemstation software. And (3) GC column: omega wax 100(15m x 0.1.1 mm x 0.1um) column. The results were converted to weight% by internal standard reference.

Fig. 3 depicts the distribution of FFA, MAG, DAG and TAG in Ensure and the GBFS products of the present disclosure. The percentage of FFA, MAG, DAG and TAG in the oil was determined by thin layer chromatography.

Example 3: another example of a process to make a MAG-rich product compared to the starting TAG-rich oil.

The block flow diagram in fig. 4 depicts the following procedure.

The vegetable oil was added to citric acid and sodium hydroxide (caustic) in DI water and heated to 33 ℃ +/-2 ℃. Degassing was performed using a low vacuum to remove oxygen. Lipase AY was added. The mixture was hydrolyzed under a blanket of nitrogen at a temperature of 33 deg.C +/-2 deg.C for 14-24 hours.

The lipase was inactivated at 70 ℃ +/-2 ℃ for 1 hour. The aqueous phase with the inactivated enzyme is discharged. Glycerol was added. The reaction was then cooled to 22 ℃ +/-2 ℃. Lipase G was then added and the water was evaporated under moderate vacuum.

Re-esterification was carried out at 20 deg.C +/-2 deg.C under high vacuum (about 720mmHg) for 72 hours. Lipase G was inactivated at 70 ℃ +/-2 ℃ for one hour, and salt was added to the reaction.

Enzyme inactivation and phase separation were carried out at 70 ℃ +/-2 ℃ for 1 hour under a blanket of nitrogen. The aqueous phase containing the inactivated enzymes, glycerol and salts is discharged. The reaction was cooled to 60 ℃ +/-2 ℃. Adding an antioxidant. The final product was stored under nitrogen at 4 ℃ +/-2 ℃.

Example 4: ready-to-drink formulation comprising MAG and hydrolyzed protein

The products of the present disclosure are produced in conventional "milkshake" formulations that contain sources of fat, protein, carbohydrate, vitamins and fiber in addition to conventional surfactants and stabilizers typically found in these products. Each 250 ml. The ingredients that will appear on the ingredient label are as follows: water, organic agave syrup, hydrolyzed pea protein, hydrolyzed blend oil, Arabic gum, sunflower lecithin, xanthan gum, oligosaccharide, potassium sorbate, sodium benzoate, instant coffee, natural organic vanilla essence, vitamin C, vitamin E succinate, vitamin A palmitate, nicotinamide, D-calcium pantothenate, pyridoxine hydrochloride, thiamine hydrochloride, riboflavin, vitamin D3, folic acid, cyanocobalamine and vitamin K2.

Carbohydrates are supplied from fruit and agave syrups in the form of monosaccharides (glucose and fructose).

The protein in the product is partially hydrolyzed pea protein (PURIS pea protein 870H, World Food Processing LLC, Turtle Lake, WI 54889).

We also produced a large amount of hydrolysed pea protein (EHP) in which the size of the peptides was in a range more suitable for biological transport across the intestinal wall. EHP is produced by further enzymatic hydrolysis. For example, partially hydrolysed pea protein (Puris pea protein 870H as described above) was dissolved in 100mM phosphate buffer to a concentration of 25 mg/ml. The enzyme was added and the reaction was incubated overnight at 50 ℃. Three different commercial GRAS enzymes were evaluated: alkaline protease, thermophilic enzyme and flavor enzyme.

Size exclusion chromatography was used to assess the average size and distribution of peptides in protein samples. The samples were dissolved in 100mM phosphate buffer pH6.8 to a concentration of 25mg/mL and analyzed on a Shimadzu HPLC with UV detector (214nm) using a Phenomenex Yarra 3um SEC-2000 column (diluted with 100mM sodium phosphate buffer pH6.8, flow rate 0.8mL/min at room temperature). The samples were compared to molecular weight standards (Phenomenex SEC standard part ALO-3042). The size was estimated using a calibration curve generated from a known molecular weight standard. The average size of the unhydrolysed pea protein is about 200 amino acids. The average size of the Puris pea 870H partially hydrolysed material was 34 amino acids, a considerable fraction of which was in the range of 2-40 amino acids, similar to other (whey-based) protein hydrolysates.

Fig. 5 depicts e.phillips et al 2005 "peptide-based formulation: enteral feeding of nutritional medications? (Peptide-Based forms: the nutritional of Industrial Feeding: 40-45, the distribution of amino acids and peptides in ready-to-drink nutritional beverages. FIG. 5 depicts the percentage of amino acids in the products of the present disclosure that are Peptamen, Pertive, Crucial, Pivot, and GBFS (X axis) that are 1 amino acid, 2-4 amino acids, 9-10 amino acids, 10-40 amino acids, or greater than 40 amino acids (Y axis). GBFS EHP has an average size of 3-4 amino acids, mostly 1-7 amino acids.

Fat is provided in the form of the reconstituted lipid MAG and is made from a mixture of olive oil (70%), sunflower oil (21%) and linseed oil (9%), providing the energy and other benefits of polyunsaturated fatty acids (PUFAs), omega-6 PUFAs and omega-3 PUFAs. The omega-6/omega-3 ratio is about 4/1.

Gum arabic, sunflower lecithin and xanthan gum are common GRAS food ingredients used to provide structure and stability to beverages. Oligosaccharides provide indigestible fiber. Potassium sorbate and sodium benzoate are common food preservatives. Instant coffee and vanilla provide flavor.

Comprising a vitamin pack containing fat-soluble and water-soluble vitamins.

Table 3 shows prototype product nutritional labels. In Table 3,% per dayThe ratio (% DV) is based on a 2,000 calorie diet.

The Daily Value (DV) was not determined.

And (4) sensory evaluation. The taste panel found that FFAs produced from the vegetable oil mixture did not eat well. Surprisingly, the MAG oil produced with this mixture was palatable and similar in mouthfeel and texture to the original triglyceride oil. When MAG oil is formulated into the RTDS product described above, its flavor is acceptable and indistinguishable from similar commercial products containing intact (undigested) lipids and proteins.

Example 5: production of multiple batches of EMO

The time for the process described in example 1 for making the EMO was evaluated to produce an oil with greater than 70% MAG, greater than > 85% MAG + FFA, and a TAG content of 5% or less.

Table 4 depicts scans of TLC plates showing the concentration of MAG and FFA in enzyme modified EMO produced from olive oil/linseed oil/sunflower oil mixture (ratio 7/2/1). In these experiments, step 2 was extended to 84 hours to determine the upper time and temperature limits to avoid TAG formation.

Experiment of	TAG	DAG	MAG	FFA	FFA+MAG
						1	4	26	57	13	70
2	4	11	72	13	85
						3	5	10	76	9	85

The reaction can be monitored by TLC analysis in substantially real time and stopped at any time point during step 2 to give the desired amounts of MAG, DAG, TAG and FFA. The 72 hour reaction time of step 2 was found to be a practical and fruitful stop and is shown in example 6.

Example 6: enzyme modified almond oil without TAG was evaluated by NMR analysis.

Almond oil was used to produce TAG-free EMO. The reaction conditions were as described in example 1. Step 2 was 72 hours to minimize TAG production.

FIGS. 6A-6B depict EMO produced from almond oil¹³C-NMR analysis. FIG. 6A shows that the TAG (glyceryl tristearate) is associated with a real TAG¹³C-NMR signals and characteristic peaks at 62.173ppm and 68.921 ppm. FIG. 6B shows EMO¹³C-NMR signal. It indicates no discernable TAG signal. Integration of the actual signal showed an acylglycerol distribution between MAG: DAG of 88%: 12%. Samples were analysed using a JEOL ECA600II type NMR spectrometer operating at 600MHz protons and 150MHz carbon. Samples were made in 5mm tubes and run locked with CDCl3 at ambient temperature.

Example 7: clinical testing of EMO-based Ready-to-drink milkshakes

Studies have been designed to show that patients with EPI can consume food produced with our TAG-free oils without taking enzyme supplements, but still take up lipids and produce TAG in their serum.

The clinical study is a single-center, randomized, double-blind, cross-over trial evaluating the blood lipid levels, safety, tolerability, and palatability of an EMO-based ready-to-drink milkshake (RTDS) compared to standard nutritional supplements used with pancreatin replacement therapy capsules ("PERT").

Inclusion criteria for this study included:

1. an informed consent is provided which signs and notes the date.

2. Indicating a willingness to follow all study procedures and to be available at any time during the study.

3. Male or female, 12 years old or older.

Diagnosis of CF

5. Currently, commercial pancreatin products are being treated for more than 3 months.

6. The clinical condition is stable within 1 month after recruitment, and no acute respiratory disease signs exist.

7. Defined as stable body weight that did not drop more than 5% within 3 months after enrollment.

8. Women with fertility potential should agree to continue to use the medically acceptable method of birth control throughout the study and within 30 days after the last study medication. Medically acceptable method of birth controlComprises bilateral tubal ligation, or use of contraceptive implant, contraceptive injection (Depo-Provera)^TM) Intrauterine devices or oral contraceptives taken within the last 3 months (if the subject agrees to continue use during the study or to use another method of birth control), or a double barrier method consisting of a combination of any two of the following methods: diaphragm, cervical cap, condom or spermicide.

9. Oral medications and oral nutritional supplements can be taken and compliance with study intervention is desirable.

10. Agreement to follow lifestyle precautions throughout the study (see section 5.3).

Exclusion criteria for this study included:

1. there are cardiovascular, respiratory (except for underlying disease), urogenital, gastrointestinal/hepatic (except for underlying disease), hematological/immunological, head, ear, eye, nose, throat, dermatological/connective tissue, musculoskeletal, metabolic/nutritional (except for underlying disease), endocrine (except for controlled diabetes), neurological/psychiatric, milk, nut or soybean allergies, recent major surgery or medical history, physical examinations and/or laboratory assessments revealing evidence of other related diseases that may limit participation in or the completion of the study.

2. History of acute abdomen disease in the past year.

3. History of fibrotic colon disease.

4. There was a history of distal ileus syndrome (DIOS) within 6 months prior to enrollment.

5. Solid organ transplantation or surgery affecting the large intestine (except appendectomy).

6. Small bowel surgery (e.g., gastrectomy or pancreatectomy) that significantly affects absorptive capacity.

7. Inflammatory bowel disease, including chronic diarrhea disease not associated with pancreatic insufficiency.

8. Celiac disease or crohn's disease.

9. Receiving enteral tube feeding of greater than or equal to 50% of daily caloric intake.

10. Pregnancy or lactation.

11. Any type of malignancy affecting the digestive tract over the last 5 years.

12. It is known to be allergic to pancreatin or to inactive ingredients (excipients) of pancreatin capsules.

13. Suspected noncompliance or noncompliance.

14. Test drug intake was within 30 days before study initiation.

15. The investigator considered a mental disability or any other lack of health that prevented the subject from participating in or being able to complete the study.

16. History diagnosis of human immunodeficiency virus.

17. Forced Expiratory Volume (FEV) for lung transplantation or other solid organ transplantation or recording is listed as ≦ 25%.

18. Using lipid lowering therapy, a statin, a fibrate, niacin, and a proprotein convertase subtilisin kexin type 9(PCSK9) inhibitor, could not be withheld for at least 14 days and 15 days prior to study day 1. The patient is male or female, 12 years of age or older, diagnosed with cystic fibrosis. Patients are also currently treated with commercial pancreatin products for more than 3 months and are stable clinically within 1 month after enrollment with no evidence of acute respiratory disease.

Patients came to the clinic after overnight fasting and a standardized dinner. Those in arm 1 (10 patients) took RTDS with PERT placebo and those in arm 2 (10 patients) administered a standard nutritional supplement containing PERT. Blood samples were taken from patients in both study arms continuously over 6 hours (0, 1,2, 3, 4, 5, 6 hours) without repeated administration of RTDS or standard nutritional supplements. Water may be drunk during the study.

Patients returned to clinic for treatment 2 after overnight fasting and a standardized dinner (cross-treatment). Patients in arm 1 were administered a standard nutritional supplement containing PERT, and patients in arm 2 were administered RTDS and PERT placebo. Blood samples were taken from patients in both study arms continuously over 6 hours (0, 1,2, 3, 4, 5, 6 hours) without repeated administration of RTDS or standard nutritional supplements.

The standard nutritional supplement contains: water, glucose syrup, sugar, vegetable oils (canola, high oleic sunflower, corn), milk protein concentrate and less than 2% soy protein isolate, calcium caseinate, sodium caseinate, acacia gum, fructo-oligosaccharides, inulin (from chicory), soy lecithin, salt, natural and artificial flavors, carrageenan potassium citrate, calcium phosphate, magnesium chloride, sodium ascorbate, choline bitartrate, DL-alpha-tocopheryl acetate, ascorbic acid, potassium chloride, ferrous sulfate, zinc sulfate, niacinamide, calcium pantothenate, manganese sulfate, copper sulfate, pyridoxine hydrochloride, thiamine hydrochloride, beta-carotene, vitamin a palmitate, riboflavin, folic acid, chromium chloride, biotin, potassium iodide, phytomenadione, sodium selenite, sodium molybdate, vitamin D3, vitamin B12.

The RTDS contains: water, organic jujube syrup, enzyme-modified almond oil, enzymatically hydrolyzed pea protein, soluble corn fiber, cocoa powder, natural flavors, salt, vitamin C, vitamin E (dl-alpha-tocopheryl acetate), niacinamide, pantothenic acid (D-calcium pantothenate), vitamin B6 (pyridoxine hydrochloride), vitamin a (retinyl palmitate), vitamin B2 (riboflavin), vitamin B1 (thiamine hydrochloride), L-methylfolic acid, vitamin K1, vitamin D3 (cholecalciferol), biotin, vitamin B12 (cyanocobalamin).

The nutritional supplement was 5.9 wt.% fat, 5.9 wt.% protein and 19 wt.% carbohydrate, while the RTDS was 7.4 wt.% fat, 4.7 wt.% protein and 9.8 wt.% carbohydrate.

The lipid dose was 0.5g/kg body weight. The lipids in the standard nutritional supplement are TAG mixtures from canola, high oleic sunflower oil and corn oil. In an interventional beverage, MAG is produced from almond oil. The lipid dose for each patient is based on a well-established "lipid tolerance test" which is similar in design and scope to the well-known glucose tolerance test. The recommended dose for these tests is 0.5-1.0g/kg administered over 20-30 minutes. Samples were taken at the start of the test, 6 hours per hour. Serum triglyceride levels are measured using standard laboratory methods.

The dose of PERT used in the study followed the manufacturer's recommended dosing guidelines, i.e., lipase activity was 2,500iu per gram of fat ingested. This translates into 3-4 capsules at the crossover stage of the study.

Fig. 7 depicts the rise in serum triglycerides after two patients ingested the EMO-based ready-to-drink milkshake. In fig. 7, the dotted line represents a patient receiving 0.5g/Kg body weight of the enzyme-modified almond oil, and the solid line represents a patient receiving 0.5g/Kg body weight of canola oil, high oleic sunflower oil and corn oil contained in RTDS consumed over half an hour. Figure 7 shows that the enzyme modified oil in RTDS is absorbed by the patient and converted to serum triglycerides.

Fig. 8 depicts the rise in serum triglycerides after ingestion of a test beverage by another patient. In fig. 8, the dashed line represents serum triglycerides after ingestion of the fert-free MAG-based RTDS and the solid line represents the PERT-containing RTDS standard of care. Patients received 0.5g/Kg body weight of enzymatically modified almond oil or 0.5g/Kg body weight of corn oil, incorporated into RTDS consumed over a half hour.

In this patient, lipid uptake and conversion to serum triglycerides was significantly faster following ingestion of MAG-based RTDS serum without PERT than with PERT-containing TAG-based (standard of care) products. This indicates that in addition to EPI, patients also suffer from cholestasis (disruption of intrahepatic bile flow), not just lack sufficient enzymes, and therefore do not properly emulsify canola, high oleic sunflower and corn oils in standard care beverages. The oil needs to be emulsified into very small droplets to create the surface area required for pancreatic lipase to hydrolyze the oil to MAG and FFA for transport to intestinal cells. This activity requires bile acids in the liver. TAG in standard care beverages cannot be emulsified into the microemulsion required for optimal lipase activity in the small intestine and therefore serum triglycerides are not increased as rapidly as MAG formulations which do not require lipase activity.

In all cases, the actual lipid dose per patient was the same: 0.5g/kg body weight.

Table 5 below shows the sugar intake of 65kg patients assumed in the study for standard nutritional supplements and RTDS. Most of the sugar and carbohydrates in the control beverage were glucose or starch and were readily converted to glucose. In the study beverage, it was assumed that 65kg patients would consume a smaller amount of beverage (higher lipid concentration of the study beverage) and that carbohydrates were supplied in lower amounts and as a 50:50 mixture of fructose and glucose. Therefore, less glucose is actually consumed.

Figure 9 depicts the mean blood glucose rise over a six hour sampling period for 8 patients each of arm 1 and arm 2 in the study. Patients who consume RTDS have a significant decline in post-prandial maximum blood glucose levels and remain below the standard nutritional supplement cohort. These results indicate that EMO-based nutritional supplements (rich in 1-monoacylglycerols, such as 1-oleylmonoglycerol) can have a positive effect on glucose homeostasis compared to standard nutritional supplements containing PERT, which are expected to produce only small amounts of 2-monoacylglycerols, such as 2-oleylmonoglycerol. Depending on the amount of glucose administered, the blood glucose levels in the RTDS cohort are expected to be higher and show unexpected results.

Figure 10 depicts the mean serum triglyceride levels of 8 patients each of arm 1 and arm 2 over a six hour sampling period in the study. Both the test and control groups showed similar triglyceride levels, indicating that patients absorbed the same amount of lipid from either a standard nutritional supplement containing PERT or an RTDS without PERT.

Example 8: high calorie, PERT-free ready-to-drink milkshake

High calorie RTDS can be prepared as follows: DI water (about 60% of the final volume) was added to the main mixing vessel and the water was heated to about 60 ℃. Mixing in the hydrolyzed protein, and adding agave syrup. The combination was performed using a hand mixer. In a separate container, warm (60 ℃) EMO and lecithin were combined. When the lecithin was dissolved, the EMO/lecithin was added to the aqueous phase and mixed. Additional water was added to reach the final weight (volume). Emulsifying with a shear mixer. To prepare very high calorie RTDS, higher levels of components are added.

After the beverage base is produced, various flavors, masking and blocking agents may be added to produce unique products such as chocolate, vanilla, strawberry, and the like.

The method can also be used to produce high calorie products in semi-solid form, such as pudding, mousse, "popsicle" and ice cream type products, using a beverage base formulation with the addition of viscosity modifiers such as xanthan gum and acacia gum.

Table 6 shows the main components and nutritional value of high calorie (1.5kcal/mL) RTDS.

Table 7 shows the major components and nutritional value of very high calorie (2.5kcal/mL) RDTS.

Example 9: high calorie, PERT-free bar

High heat bars can be prepared as follows: DI water (about 60% of the final volume) was added to the main mixing vessel and the water was heated to about 60 ℃. Mixing in the hydrolyzed protein, and adding agave syrup. Warm (60 ℃) EMO and lecithin were combined in separate containers with a hand mixer. When the lecithin was dissolved, the EMO/lecithin was added to the aqueous phase and mixed. Additional water was added to reach the final weight (volume).

Table 8 shows the main ingredients and nutritional value of the high calorie PERT-free bar.

After the stick base is produced, various flavors, masking and blocking agents can be added prior to baking to produce unique products such as chocolate, vanilla, and the like.

Example 10: manufacture of ready-to-drink milkshakes

DI water was added to the main mixing vessel and the water was heated to about 60+/-2 ℃. Once the water reaches 60 ℃, a syrup (such as agave or date syrup) is slowly added under low agitation. Mixing into a solution. The dry mass of each material (vitamin/mineral mixture and hydrolyzed protein) was weighed out and mixed in separate containers. The dry ingredients were mixed thoroughly. The mixed dry ingredients were slowly added directly to the main mixing vessel with sweep agitation with low agitation. The EMO was weighed out in a separate vessel. EMO was heated to 60+/-2 ℃. Sunflower lecithin was slowly added to warm EMO and mixed (as needed) with moderate stirring until the sunflower lecithin was completely mixed into the solution. The EMO/sunflower lecithin mixture was slowly added to the main mixing vessel. Distilled water was added to increase the volume to 95% of the total fluid mass and the temperature was returned to 70+/-2 ℃. The flavor and color are slowly added to the main mixing vessel. The stabilizer (such as acacia gum) is added slowly to the main mix vessel. The solution was mixed for 20 minutes (to increase the viscosity). The temperature of the solution was maintained at 70+/-2 ℃. The QS solution was adjusted to final volume with distilled water. The material was passed through a pressure drop homogenizer to produce a stable emulsion. The material is pasteurized or sterilized. The material was cooled to room temperature. The product is filled into the package.

Example 11: removal of MCPD from almond oil

Enzymatically modified almond oil was prepared as described in example 6.

Samples of the enzyme modified almond oil were sent to a third party laboratory (Eurofins Scientific inc. nutrition Analysis Center 2200Rittenhouse Street, Suite 150Des animals, IA 50321) and analyzed using standard methods as described below.

AOCS official method Cd 29b-13 (revised 2017) -2-and 3-MCPD Fatty Acid Esters and glycidyl Fatty Acid Esters in Edible Oils and Fats by Alkaline Transesterification and GC/MS (2-and 3-MCPD Fatty Acid Esters and Glycidol Fatty Acid Esters in soluble Oils and Fats by Alkaline line Trans discovery and GC/MS). The method is used for determining fatty acid esters of 2-chloropropane-1, 3-diol (2-MCPD), 3-chloropropane-1, 2-diol (3-MCPD) and glycidol in edible oil and fat. See also AOCS official methods Cd 29a-13 or Cd 29 c-13. The bound glycidol is the sum of all glycidol derivatives cleaved by base-catalyzed alcoholysis. Bound glycidol content is reported in milligrams per kilogram (mg/kg). Bound 2-MCPD is the sum of all 2-MCPD-derivatives that are cleaved by base-catalyzed alcoholysis. Bound 2-MCPD content is reported in milligrams per kilogram (mg/kg). Bound 3-MCPD is the sum of all 3-MCPD-derivatives that are cleaved by base-catalyzed alcoholysis. Bound 3-MCPD content is reported in milligrams per kilogram (mg/kg). This method describes a procedure for the parallel determination of glycidol and 2-MCPD and 3-MCPD in both bound and free form in oils and fats. The method is based on base-catalyzed ester cleavage to convert the liberated glycidol to monobromopropylene glycol (MBPD) and the derived free diols (MCPD and MBPD) and phenylboronic acid (PBA). Although free MCPD and glycidol are only present in fats and oils in low to negligible amounts, significant amounts will increase the determination of bound analyte proportionately. The process is applicable to both solid and liquid fats and oils.

And (6) obtaining the result. The total amount of 2-MCPD (free and bound) and 3-MCPD (free and bound) in the starting oil was 0.65mg/kg and 1.17mg/kg, respectively. The total 2-MCPD (free and bound) and 3-MCPD (free and bound) of the enzyme modified almond oil (example 6) were both <0.10mg/kg, below the limit of quantitation for the assay measuring MCPD (0.10 mg/kg). Thus, the sum of total MCPD detected in the starting almond oil and the enzyme-modified almond oil was 1.82mg/kg and <0.10mg/kg, respectively.

Thus, the present invention is well adapted to carry out the objects and advantages mentioned as well as those inherent therein. While numerous changes may be made by those skilled in the art, such changes are encompassed within the spirit of the invention as set forth in part in the appended claims.

Example 12: fatty acid profile in starting and MAG oils

MCT oil, canola oil, and almond oil are processed according to the manufacturing process of the present disclosure. The fatty acid profile of each oil was measured in both the starting oil and MAG oil as shown in table 9 below.

After derivatization to fatty acid methyl esters, the fatty acids were analyzed and compared to standards. For derivatization, a sample (500. mu.l) was added to a 5-ml reaction tube containing 2ml of boron trifluoride solution (12% in methanol), 20. mu.l of dimethoxypropane and 100. mu.l of tridecanoic acid internal standard solution (10 mg/ml). The reaction tube was vortexed and incubated in a60 ℃ heat block for 30 minutes. The reaction tube was removed from the heating block and allowed to cool for 15 minutes. Then, 1ml of distilled water was added to quench the reaction, and then 1ml of hexane was added. The reaction tube was vortexed for 60 seconds and the phases were allowed to separate for 3 minutes. The top (hydrophobic) phase was transferred to a 1.5-ml tube containing about 50mg sodium sulfate (anhydrous). After vortexing for 60 seconds, the 1.5-ml tube was centrifuged and approximately 500. mu.l of the clear, dried hydrophobic phase was transferred to a gas chromatography sample vial. Samples were analyzed using an Agilent 7890A gas chromatograph with a flame ionization detector and Agilent Openlab CDS Chemstation software. And (3) GC column: omega wax 100(15m x 0.1.1 mm x 0.1um) column. The results were converted to weight% by internal standard reference.

As shown, the starting oil and MAG oil contained substantially the same fatty acid profile.

Separate experiments were performed using almond oil and canola oil to obtain the data in table 10, which show the same results for oleic, linoleic and linolenic acids.

These results indicate that the manufacturing process of the present disclosure is able to preserve the fatty acid profile of the starting oil in MAG oil. This result is different from conventional "distilled" oils, which have a fatty acid profile that is significantly different from the starting oil based on the functional use of the distilled oil.

These results also indicate that the processing oils of the present disclosure can be used to provide "complete nutrition" by providing linoleic and linolenic acids. Similar results are expected for oils containing high EPA and DHA, such as fish oil and algae oil, to provide a complete nutritional formulation.

Example 13: EMO Process preserves essential fatty acids found in the original oil

Canola EMO was produced as described in example 1. Fatty acids were measured as described in example 2. The results of the fatty acid analysis are shown in fig. 11. Canola oil and EMO produced from canola oil maintain the integrity of the fatty acid profile and comprise the essential fatty acids linoleic and linolenic acids. Similar results were obtained with EMO in olive oil, sunflower oil and almond oil. Commercial "glyceryl monostearate" (Profood Products, Naperville, IL) contains little or no unsaturated fatty acids, whereas "Capmul GMO 50" (Abitec Corporation, Janesville, WI) contains primarily the monosaturated fatty acid oleic acid, little linoleic acid, and virtually no linolenic or eicosene (C20:1) fatty acids.

Example 14: preserving beneficial compounds

Vegetable oils contain many lipids, steroids, esters and phenolic compounds in trace amounts that contribute to the taste and health benefits of the oil. Many of these compounds are preserved or enriched during EMO. This is shown in fig. 12A-12B. Canola EMO was prepared as described in example 1. Lipid samples were derivatized with 3-picolamide for LC/MS/MS analysis. Briefly, 150. mu.L of standard (0.1-100. mu.g/mL) and 20. mu.L of the internal standard mixture were mixed and dried under nitrogen. To the dried residue was added 200 μ L oxalyl chloride (2M in dichloromethane) and the mixture was incubated on a heat block for 5min at 65 ℃ then dried under nitrogen. To the residue was added 150. mu.L of 3-picolinamide (1% in acetonitrile, v/v) to form 3-picolinamide (FA-PA). The mixture was incubated at room temperature for 5min and then dried under nitrogen to give derivatized FA. The dried FA derivative was dissolved in 1000 μ L of ethanol and further diluted 10-fold with ethanol before LC-MS analysis. LC-MS analysis was in ionization mode: positive (total fatty acids and lipidomics) and negative (lipidomics) ionization. Software for analysis: thermo Scientific Freestyle&LipidSearch. The Orbitrap fusion method: data dependent acquisition MS²(total fatty acids) and MS.

The retained or enhanced compound comprises: glycosphingolipids (e.g., ceramide phosphate), glycosylglycerolipids (e.g., monogalactosyl glycerol), phosphatidylcarbinols, steroids (e.g., sitosterol esters), neutral lipids (e.g., campesterol esters), sphingolipids, phosphatidylglycerols, wax esters, and sphingomyelins. Coenzymes have also been found to be enhanced.

These compounds do not remain in the commercial (distilled) monoglyceride product.

In a first embodiment, the food product comprises an oil and has a total caloric content of from about 25kcal to about 1,000kcal, wherein from about 5% to about 75% of the total caloric content is derived from the oil, and wherein the oil comprises less than 10% by weight Triacylglyceride (TAG) based on the total weight of the oil, wherein the oil is substantially free of Monochloropropanediol (MCPD).

In one aspect of the first embodiment, the oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the oil.

In another aspect of the first embodiment, the oil includes greater than 60 wt.% MAG based on the total weight of the oil.

In another aspect of the first embodiment, the oil includes greater than 70 wt.% MAG based on the total weight of the oil.

In another aspect of the first embodiment, the oil includes greater than 80 wt.% MAG based on the total weight of the oil.

In another aspect of the first embodiment, the oil includes greater than 90 wt MAG based on the total weight of the oil.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, about 10% to about 60% of the total caloric content is derived from the oil.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, about 20% to about 50% of the total caloric content is derived from the oil.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, about 25% to about 45% of the total caloric content is derived from the oil.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, about 30% to about 40% of the total caloric content is derived from the oil.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product further comprises a source of carbohydrates.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the food product further comprises a carbohydrate source, and the carbohydrate source comprises fruit or agave syrup.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product further comprises a carbohydrate source, and the carbohydrate source comprises a monosaccharide.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product further comprises a carbohydrate source, and from about 20% to about 50% of the calories originate from the carbohydrate source.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product comprises a carbohydrate source and further comprises a protein source.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product comprises a carbohydrate source and further comprises a protein source, and from about 10% to about 50% of the calories are derived from the protein source.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product comprises a carbohydrate source and further comprises a protein source, and the protein source comprises hydrolyzed or partially hydrolyzed protein.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the food product comprises a carbohydrate source and further comprises a protein source, and the protein source comprises hydrolyzed or partially hydrolyzed protein, wherein the hydrolyzed protein is selected from the group consisting of hydrolyzed pea protein and whey-based hydrolysate.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the food product further comprises a protein source.

In another aspect of the first embodiment or any of the preceding aspects of the first embodiment, the food product further comprises a protein source, and from about 10% to about 50% of the calories originate from the protein source.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the food product further comprises a protein source, wherein the protein source comprises hydrolyzed or partially hydrolyzed protein.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the food product further comprises a protein source, wherein the protein source comprises hydrolyzed or partially hydrolyzed protein, and wherein the hydrolyzed protein is selected from the group consisting of hydrolyzed pea protein and whey-based hydrolysate.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the oil is a process oil derived from an oil source.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the oil is a process oil derived from an oil source, wherein the process oil comprises non-oil components derived from the oil source and naturally present in the oil source such that the non-oil components are not added to the process oil.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the oil is a process oil derived from an oil source, wherein the process oil comprises non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, and wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the oil is a process oil derived from an oil source, wherein the process oil comprises non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, and wherein the antioxidants are tocopherols.

In another aspect of the first embodiment or any preceding aspect of the first embodiment, the oil is a process oil derived from an oil source, wherein the process oil comprises non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, wherein the antioxidants are tocopherols, and wherein the tocopherols are selected from the group consisting of alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol, and gamma-tocotrienol.

In a second embodiment, a food product comprises a processing oil, a carbohydrate source, and a protein source, and has a total weight of about 25 grams to about 500 grams and a caloric density of about 1 kcal/gram to about 5 kcal/gram, wherein processing oil constitutes about 10% to about 50% of the total caloric content, and wherein the processing oil has a MAG content equal to or greater than 40 wt% based on the total weight of the processing oil and a TAG content equal to or less than 10 wt% based on the total weight of the processing oil, and wherein the processing oil has an MCPD of less than 0.10 mg/kg.

In a third embodiment, a product comprises a processing oil derived from an oil source, wherein the processing oil comprises a MAG content equal to or greater than 40 wt% of the total weight of the processing oil, wherein the processing oil is free of TAG or comprises a TAG content equal to or less than 10 wt% of the total weight of the processing oil, and wherein the processing oil comprises non-oil components derived from the oil source and naturally present in the oil source such that the non-oil components are not added to the processing oil, wherein the oil source comprises from about 1.00mg/kg to about 12.00mg/kg MCPD, and wherein the processing oil comprises less than 0.100mg/kg MCPD.

In one aspect of the third embodiment, the oil source is from a source selected from the group consisting of plants, animals, algae, and fish.

In another aspect of the third embodiment, the oil source is of plant origin.

In another aspect of the third embodiment, the oil source is selected from the group consisting of olive oil, sunflower oil, corn oil, almond oil, rapeseed oil, palm oil, soybean oil, linseed oil, and mixtures thereof.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, and the antioxidants are tocopherols.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, the antioxidants are tocopherols, and the tocopherols are selected from the group consisting of alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol, and gamma-tocotrienol.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the processing oil comprises a MAG content of about 50 wt% to about 95 wt%, based on the total weight of the processing oil.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the processing oil comprises a MAG content of about 50 wt% to about 95 wt%, based on the total weight of the processing oil, and the processing oil comprises a TAG content of about 5 wt% to about 0.5 wt%, based on the total weight of the processing oil.

In another aspect of the third embodiment or any preceding aspect of the third embodiment, the processing oil comprises a TAG content of about 5 wt.% to about 0.5 wt.%, based on the total weight of the processing oil.

In a fourth embodiment, a method of making a monoacylglycerol-rich oil, comprising: mixing a starting oil comprising Triacylglycerol (TAG) and about 1.00mg/kg to about 12.00mg/kg MCPD, a buffer solution, and a first enzyme capable of hydrolyzing the TAG to Free Fatty Acids (FFA) to produce a first reaction mixture, wherein the amount of TAG is greater than 50% by weight of the total weight of starting oil; reacting the first reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAG for a first period of time to produce an aqueous phase and a first lipid reaction product comprising FFA; inactivating the first enzyme in the first lipid reaction product; collecting the first lipid reaction product by removing it from the aqueous phase; mixing the first lipid reaction product with food grade glycerol and a second enzyme capable of esterifying FFA to form a second reaction mixture; reacting the second reaction mixture for a second period of time to produce a second lipid reaction product comprising a lipid oil phase and a glycerol phase; inactivating the second enzyme in the second lipid reaction product; adding a salt to the reaction product and separating a lipid oil phase from the glycerol phase; and collecting the lipid oil phase, wherein the lipid oil phase is substantially free of MCPD.

In one aspect of the fourth embodiment, the starting oil is an oil derived from a plant, animal or fish.

In another aspect of the fourth embodiment, the starting oil is a vegetable oil or a mixture of vegetable oils.

In another aspect of the fourth embodiment, the starting oil is a vegetable oil selected from the group consisting of olive oil, sunflower oil, corn oil, almond oil, rapeseed oil, palm oil, soybean oil, linseed oil, and mixtures thereof.

In another aspect of the fourth embodiment, the first enzyme is a lipase.

In another aspect of the fourth embodiment, the first enzyme is lipase AY.

In another aspect of the fourth embodiment, the buffer solution is a sodium citrate solution.

In another aspect of the fourth embodiment, the first period of time is a period of time sufficient to hydrolyze at least 94% of the TAGs in the starting oil.

In another aspect of the fourth embodiment, the first period of time is between about 14 hours and about 24 hours.

In another aspect of the fourth embodiment, said step of reacting said reaction mixture under conditions sufficient for said first enzyme to hydrolyze said TAG is carried out at a temperature between about 30 ℃ to about 35 ℃.

In another aspect of the fourth embodiment, the steps of mixing a starting oil comprising Triacylglycerols (TAGs), a buffer solution, and a first enzyme capable of hydrolyzing the TAGs to Free Fatty Acids (FFAs) and reacting the reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAGs to FFAs are conducted under a nitrogen atmosphere.

In another aspect of the fourth embodiment, the second enzyme is a lipase.

In another aspect of the fourth embodiment, the second enzyme is lipase G.

In another aspect of the fourth embodiment, the second time period is a time period sufficient to result in an enrichment of MAG in the lipid oil phase of about 60% to 95%.

In another aspect of the fourth embodiment, the second period of time is between about 24 hours and about 72 hours.

In another aspect of the fourth embodiment, said step of reacting said second reaction mixture for a second period of time to produce a lipid oil phase and a glycerol phase is conducted at a temperature between about 17 ℃ and 23 ℃.

In another aspect of the fourth embodiment or any of the preceding aspects of the fourth embodiment, the method further comprises drying the second lipid reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the second lipid reaction product.

In another aspect of the fourth embodiment or any of the preceding aspects of the fourth embodiment, the method further comprises drying the second lipid reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the second lipid reaction product, wherein the step of drying the second lipid reaction product is performed at a temperature between 20 ℃ and 30 ℃.

In another aspect of the fourth embodiment or any of the preceding aspects of the fourth embodiment, the method further comprises drying the second lipid reaction product by applying a vacuum for a third period of time sufficient to remove at least a portion of the water from the second lipid reaction product, wherein the drying step is performed over the second period of time.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the step of inactivating the second enzyme is performed by heating the second lipid reaction product.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the step of inactivating the second enzyme is performed by heating the second lipid reaction product, wherein the heating is performed at a temperature of at least 70 ℃ for at least 1 hour.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the step of separating the lipid oil phase from the glycerol phase comprises adding sodium chloride to the second lipid reaction product.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the step of separating the lipid oil phase from the glycerol phase comprises adding sodium chloride to the second lipid reaction product, wherein the final concentration of sodium chloride comprises at most 0.3% by weight sodium chloride.

In another aspect of the fourth embodiment or any of the preceding aspects of the fourth embodiment, the method further comprises reconstituting a nitrogen atmosphere on the first lipid reaction product prior to mixing the first lipid reaction product with food-grade glycerol and a second enzyme capable of esterifying FFA and glycerol.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the method further comprises adding tocopherol to the lipid oil phase after collecting the lipid oil phase.

In another aspect of the fourth embodiment or any preceding aspect of the fourth embodiment, the lipid oil phase comprises MAG in an amount from about 40 wt% to about 99 wt%, based on the total weight of the lipid oil phase, and wherein the lipid oil phase does not contain TAG or comprises TAG in an amount from about 0.1 wt% to about 10 wt%, based on the total weight of the lipid oil phase.

In a fifth embodiment, a process oil comprises Monoglycerol Oleate (MOG) and has an overall fatty acid content, wherein the MOG comprises from about 5% to about 75% by weight of the overall fatty acid content of the process oil.

In one aspect of the fifth embodiment, the process oil comprises less than 10 wt% Triacylglycerides (TAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment, the process oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment, the process oil comprises greater than 60 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment, the process oil comprises greater than 70 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment, the process oil comprises greater than 80 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment, the process oil comprises greater than 90 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is a process oil derived from an oil source.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is a process oil derived from an oil source, and the process oil includes non-oil components derived from the oil source and naturally present in the oil source such that the non-oil components are not added to the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is a process oil derived from an oil source, and the process oil includes non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is a process oil derived from an oil source, and the process oil includes non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, and wherein the antioxidants are tocopherols.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is a process oil derived from an oil source, and the process oil comprises non-oil ingredients derived from the oil source and naturally present in the oil source such that the non-oil ingredients are not added to the process oil, wherein the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof, wherein the antioxidants are tocopherols, and wherein the tocopherols are selected from the group consisting of alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol, and gamma-tocotrienol.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil is substantially free of Monochloropropanediol (MCPD).

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil has less than 0.10mg/kg Monochloropropanediol (MCPD).

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the oil source is from a source selected from a plant, an animal, algae, or fish.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the oil source is of plant origin.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the oil source is selected from the group consisting of safflower oil, grape oil, silybum oil, sesame oil, sunflower oil, wheat germ oil, pumpkin seed oil, sesame oil, rice bran oil, almond oil, rapeseed oil, peanut oil, olive oil, and coconut oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the processing oil comprises a MAG content of about 50 wt% to about 95 wt%, based on the total weight of the processing oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the processing oil comprises a TAG content of about 5 wt.% to about 0.5 wt.%, based on the total weight of the processing oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the MOG in the processing oil comprises at least 50 wt.% of 1-oleyl monoglycerides of the total MOG.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the MOG in the processing oil comprises at least 60 wt.% of 1-oleyl monoglycerides of the total MOG.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the MOG in the processing oil comprises at least 70 wt.% of 1-oleyl monoglycerides of the total MOG.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the MOG in the processing oil comprises at least 80 wt.% of 1-oleyl monoglycerides of the total MOG.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil further comprises linoleic acid, wherein the linoleic acid is present in the process oil in an amount from about 1.5% to about 90% by weight of the total fatty acid content of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil further comprises linoleic acid, wherein the linoleic acid is present in the process oil in an amount from about 10% to about 90% by weight of the total fatty acid content of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil further comprises linoleic acid, wherein the linoleic acid is present in the process oil in an amount from about 20% to about 90% by weight of the total fatty acid content of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil further comprises linoleic acid, wherein the linoleic acid is present in the process oil in an amount from about 10% to about 25% by weight of the total fatty acid content of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, the process oil further comprises linolenic acid, wherein the linolenic acid is present in the process oil in an amount of from about 0.01% to about 2% by weight of the total fatty acid content of the process oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linoleic acid in the process oil is between about 0.01 and about 5.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linoleic acid in the process oil is between about 1 and about 4.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linoleic acid in the process oil is between about 3 and about 4.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linolenic acid in the processing oil is between about 1 and about 100.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linolenic acid in the processing oil is between about 10 and 100.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the ratio between the amounts of oleic acid and linolenic acid in the processing oil is between about 10 and 30.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the fatty acid profile of the process oil is substantially the same as the fatty acid profile of a pre-process oil from which the process oil is produced.

In one aspect of the preceding aspect, wherein the fatty acid profile comprises oleic acid, linoleic acid and linolenic acid.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 10% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in a pre-process oil from which the process oil is produced.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 1% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the processing oil comprises MAG in an amount that is three times the amount of TAG in the pre-processing oil to produce the processing oil.

In another aspect of the fifth embodiment or any preceding aspect of the fifth embodiment, wherein the process oil comprises oleic acid in the form of oleic acid monoglyceride in an amount that is about the same as the amount of oleic acid in a pre-process oil from which the process oil is produced.

In a sixth embodiment, a process oil has a fatty acid profile including oleic acid, linoleic acid, and linolenic acid, wherein the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 10% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in a pre-process oil from which the process oil is produced, and wherein the process oil comprises greater than 50% by weight of Monoacylglycerides (MAG), based on the total weight of the process oil.

In a seventh embodiment, a process oil has a fatty acid profile comprising oleic acid, linoleic acid, and linolenic acid, wherein the amount of oleic acid, linoleic acid, and linoleic acid in the process oil is within about 1% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in a pre-process oil from which the process oil is produced, and wherein the process oil comprises greater than 50% by weight of Monoacylglycerides (MAG), based on the total weight of the process oil.

In an eighth embodiment, a processing oil comprises oleic acid and linoleic acid, wherein the ratio of oleic acid to linoleic acid in the processing oil is between about 0.01 and about 5, and wherein the processing oil comprises greater than 50 wt.% of Monoacylglycerides (MAG), based on the total weight of the processing oil.

In one aspect of the eighth embodiment, the ratio of oleic acid to linoleic acid in the process oil is between about 1 to about 4.

In another aspect of the eighth embodiment, the ratio of oleic acid to linoleic acid in the process oil is between about 3 and about 4.

In another aspect of the eighth embodiment or any preceding aspect of the eighth embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linolenic acid in the process oil is between about 1 to about 100.

In another aspect of the eighth embodiment or any preceding aspect of the eighth embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linolenic acid in the process oil is between about 10 to about 100.

In another aspect of the eighth embodiment or any preceding aspect of the eighth embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linolenic acid in the process oil is between about 10 to about 30.

In a ninth embodiment, a processing oil comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid in the processing oil is between about 1 to about 100, and wherein the processing oil comprises greater than 50% by weight Monoacylglycerides (MAG) based on the total weight of the processing oil.

In a tenth embodiment, a processing oil comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid in the processing oil is between about 10 to about 100, and wherein the processing oil comprises greater than 50% by weight Monoacylglycerides (MAG), based on the total weight of the processing oil.

In an eleventh embodiment, a processing oil comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid in the processing oil is between about 10 to about 30, and wherein the processing oil comprises greater than 50% by weight Monoacylglycerides (MAG), based on the total weight of the processing oil.

In one aspect of the ninth, tenth or eleventh embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linoleic acid in the process oil is between about 0.01 and about 5.

In one aspect of the ninth, tenth or eleventh embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linoleic acid in the process oil is between about 1 to about 4.

In one aspect of the ninth, tenth or eleventh embodiment, the process oil further comprises linoleic acid, wherein the ratio of oleic acid to linoleic acid in the process oil is between about 3 and about 4.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, at least a portion of the oleic acid is present as oleic acid Monoglyceride (MOG).

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, at least a portion of the oleic acid is present as oleic acid Monoglyceride (MOG) and is a 1-oleyl monoglyceride to at least 50 wt% of the total MOG.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, at least a portion of the oleic acid is present as oleic acid Monoglyceride (MOG) and is a 1-oleyl monoglyceride in an amount of at least 60 wt% of the total MOG.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, at least a portion of the oleic acid is present as oleic acid Monoglyceride (MOG) and is a 1-oleyl monoglyceride to at least 70 wt% of the total MOG.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, at least a portion of the oleic acid is present as oleic acid Monoglyceride (MOG) and is a 1-oleyl monoglyceride that is at least 80% by weight of the total amount of MOG.

In one aspect of the eighth, ninth, tenth, or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth, or eleventh embodiment, the process oil comprises less than 10 wt.% Triacylglyceride (TAG) based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises greater than 60 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises greater than 70 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises greater than 80 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises greater than 90 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil is derived from an oil source.

In one aspect of the foregoing aspect, the process oil comprises non-oil components that originate from and are naturally present in the oil source such that the non-oil components are not added to the process oil.

In one aspect of the preceding aspect, the non-oil ingredients are selected from the group consisting of antioxidants, vitamins, and mixtures thereof.

In one aspect of the preceding aspect, the antioxidant is tocopherol.

In one aspect of the preceding aspect, the tocopherol is selected from the group consisting of alpha-tocopherol, beta-tocopherol, delta-tocopherol, gamma-tocopherol, alpha-tocotrienol, beta-tocotrienol, delta-tocotrienol, and gamma-tocotrienol.

In an aspect of the eighth, ninth, tenth, or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth, or eleventh embodiment, the process oil is substantially free of Monochloropropanediol (MCPD).

In an aspect of the eighth, ninth, tenth, or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth, or eleventh embodiment, the process oil has less than 0.10mg/kg Monochloropropanediol (MCPD).

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the oil source is from a source selected from the group consisting of plants, animals, algae or fish.

In one aspect of the preceding aspect, the oil source is of plant origin.

In one aspect of the preceding aspect, the oil source is selected from the group consisting of safflower oil, grape oil, silybum oil, sesame oil, sunflower oil, wheat germ oil, pumpkin seed oil, sesame oil, rice bran oil, almond oil, rapeseed oil, peanut oil, olive oil and coconut oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the processing oil comprises a MAG content of about 50 wt.% to about 95 wt.%, based on the total weight of the processing oil.

In one aspect of the eighth, ninth, tenth, or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth, or eleventh embodiment, the processing oil comprises from about 5 wt.% to about 0.5 wt.% TAG content, based on the total weight of the processing oil.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the fatty acid profile of the process oil is substantially the same as the fatty acid profile of a pre-process oil from which the process oil is produced.

In one aspect of the preceding aspects, the fatty acid profile comprises oleic acid, linoleic acid, and linolenic acid.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the amount of oleic acid, linoleic acid and linolenic acid in the process oil is within about 10% of the amount of oleic acid, linoleic acid and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the amount of oleic acid, linoleic acid and linolenic acid in the process oil is within about 1% of the amount of oleic acid, linoleic acid and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the processing oil comprises MAG in an amount that is three times the amount of TAG in the pre-processing oil relative to the amount of TAG in the pre-processing oil from which the processing oil is produced.

In one aspect of the eighth, ninth, tenth or eleventh embodiment and any preceding aspect of the eighth, ninth, tenth or eleventh embodiment, the process oil comprises oleic acid in the form of oleic acid monoglycerides in an amount that is about the same as the amount of oleic acid in a pre-process oil from which the process oil is produced.

In a twelfth embodiment, the process oil comprises oleic acid and linoleic acid and has an overall fatty acid content, wherein the linoleic acid is present in an amount of from about 10 wt.% to about 90 wt.% of the overall fatty acid content of the process oil, and wherein the process oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment, the linoleic acid is present in an amount from about 20% to about 90% by weight of the total fatty acid content of the processed oil.

In another aspect of the twelfth embodiment, the linoleic acid is present in an amount from about 10% to about 25% by weight of the total fatty acid content of the processed oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the processed oil further comprises linolenic acid, wherein the linolenic acid is present in an amount of about 0.01% to about 2% by weight of the total fatty acid content of the processed oil.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linoleic acid in the process oil is between about 0.01 and about 5.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linoleic acid in the process oil is between about 1 to about 4.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linoleic acid in the process oil is between about 3 and about 4.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linolenic acid in the processing oil is between about 1 to about 100.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linolenic acid in the processing oil is between about 10 to about 100.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the ratio of oleic acid to linolenic acid in the processing oil is between about 10 to about 30.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, wherein at least a portion of the oleic acid is present as glycerol Monooleate (MOG).

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the MOG in the processing oil comprises at least 50 wt.% of 1-oleyl monoglyceride of the total amount of MOG.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the MOG in the processing oil comprises at least 60% by weight of 1-oleyl monoglyceride of the total amount of MOG.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the MOG in the processing oil comprises at least 70 wt.% of 1-oleyl monoglyceride, based on the total amount of MOG.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the MOG in the processing oil comprises at least 80 wt.% of 1-oleyl monoglyceride, based on the total amount of MOG.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises less than 10 wt.% Triacylglycerides (TAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises greater than 60 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises greater than 70 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises greater than 80 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil comprises greater than 90 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the process oil is a process oil derived from an oil source.

In one aspect of the preceding aspect, the antioxidant is tocopherol.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil is substantially free of Monochloropropanediol (MCPD).

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the process oil has less than 0.10mg/kg Monochloropropanediol (MCPD).

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the oil source is from a source selected from a plant, an animal, algae, or fish.

In one aspect of the preceding aspect, the oil source is of plant origin.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the processing oil comprises a MAG content of about 50 wt.% to about 95 wt.%, based on the total weight of the processing oil.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the processing oil comprises a TAG content of about 5 wt.% to about 0.5 wt.%, based on the total weight of the processing oil.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the fatty acid profile of the process oil is substantially the same as the fatty acid profile of a pre-process oil from which the process oil is produced.

In one aspect of the preceding aspect, the fatty acid profile comprises oleic acid, linoleic acid and linolenic acid.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 10% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 1% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in the pre-process oil from which the process oil is produced.

In one aspect of the twelfth embodiment or any preceding aspect of the twelfth embodiment, the processing oil comprises substantially the same amount of MAG relative to the amount of TAG in the pre-processing oil from which the processing oil is produced.

In one aspect of the twelfth embodiment or any of the preceding aspects of the twelfth embodiment, the process oil comprises oleic acid in the form of oleic acid monoglyceride in an amount that is about the same as the amount of oleic acid in a pre-process oil from which the process oil is produced.

In a thirteenth embodiment, a process oil has a fatty acid profile including oleic acid, linoleic acid, and linolenic acid, wherein the amount of oleic acid, linoleic acid, and linolenic acid in the process oil is within about 10% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in a pre-process oil from which the process oil is produced, and wherein the process oil comprises greater than 50% by weight of Monoacylglycerides (MAG), based on the total weight of the process oil.

In a fourteenth embodiment, a process oil has a fatty acid profile including oleic acid, linoleic acid, and linolenic acid, wherein the amount of oleic acid, linoleic acid, and linoleic acid in the process oil is within about 1% of the amount of oleic acid, linoleic acid, and linolenic acid, respectively, in a pre-process oil from which the process oil is produced, and wherein the process oil comprises greater than 50% by weight of Monoacylglycerides (MAG), based on the total weight of the process oil.

In a fifteenth embodiment, a method for promoting glucose homeostasis in a subject in need thereof, comprising: administering to the subject a composition comprising a processed oil.

In a sixteenth embodiment, a method for treating type II diabetes in a subject in need thereof, comprising: administering to the subject a composition comprising a processed oil.

In a seventeenth embodiment, a method for promoting glucose homeostasis in a subject in need thereof, comprising: administering to the subject a composition comprising a processing oil comprising oleic acid Monoglyceride (MOG), wherein at least 50% by weight of the MOG is 1-oleyl monoglyceride.

In an eighteenth embodiment, a method for treating diabetes in a subject in need thereof, comprising: administering to the subject a composition comprising a processing oil comprising oleic acid Monoglyceride (MOG), wherein at least 50% by weight of the MOG is 1-oleyl monoglyceride.

In one aspect of the seventeenth or eighteenth embodiment, at least 60% by weight of the MOG is 1-oleyl monoglyceride.

In another aspect of the seventeenth or eighteenth embodiment, at least 70% by weight of the MOG is a 1-oleyl monoglyceride.

In another aspect of the seventeenth or eighteenth embodiment, at least 80 wt.% of the MOG is a 1-oleyl monoglyceride.

In one aspect of the fifteenth, sixteenth, seventeenth, or eighteenth embodiment or any preceding aspect of the seventeenth or eighteenth embodiment, the process oil is a process oil according to any one of the fifth to fourteenth embodiments and any preceding aspect thereof.

In one aspect of the fifteenth, sixteenth, seventeenth or eighteenth embodiment or any preceding aspect of the fifteenth, sixteenth, seventeenth or eighteenth embodiment, the composition is a food product comprising a processed oil of the food product of any of the first, second or third embodiments and any preceding aspect thereof.

In one aspect of the fifteenth or seventeenth embodiment or any preceding aspect of the fifteenth or seventeenth embodiment, the subject has a disorder that affects glucose homeostasis.

In one aspect of the preceding aspects, the disorder is insulin resistance or type II diabetes.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 10 wt.% to about 75 wt.% of the total fatty acid content of the process oil.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 20 wt.% to about 75 wt.% of the total fatty acid content of the process oil.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 30% to about 75% by weight of the total fatty acid content of the process oil.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 40 wt.% to about 75 wt.% of the total fatty acid content of the process oil.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 50% to about 75% by weight of the total fatty acid content of the process oil.

In one aspect of the fifth, sixth or seventh embodiment or any preceding aspect of the fifth, sixth or seventh embodiment, the MOG comprises from about 60% to about 75% by weight of the total fatty acid content of the process oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises glycerol Monooleate (MOG) in an amount from 5% to about 75% by weight of the fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 10 to about 75 wt.% of the total fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 20% to about 75% by weight of the total fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 30% to about 75% by weight of the total fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 40% to about 75% by weight of the total fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 50 to about 75 wt.% of the total fatty acid content of the processing oil.

In one aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment or any preceding aspect of the eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiment, the processing oil comprises monoglycerides of oleic acid (MOG) in an amount from about 60 to about 75 wt.% of the total fatty acid content of the processing oil.

In a nineteenth embodiment, a process oil has a fatty acid profile, wherein the fatty acid profile of the process oil is substantially the same as a pre-process oil from which the process oil is produced, and wherein the process oil comprises greater than 50% by weight Monoacylglycerides (MAG) based on the total weight of the process oil.

In a twentieth embodiment, a food product comprises the processing oil of any one of the preceding aspects of the fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiments or the fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth embodiments.

In a twenty-first embodiment, a process oil comprises greater than 50 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil, and has an overall fatty acid content.

In a twenty-second embodiment, a process oil comprises greater than 60 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil, and has an overall fatty acid content.

In a twenty-third embodiment, a process oil comprises greater than 70 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil, and has an overall fatty acid content.

In a twenty-fourth embodiment, a process oil comprises greater than 80 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil, and has an overall fatty acid content.

In a twenty-fifth embodiment, a process oil comprises greater than 90 wt.% Monoacylglycerides (MAG), based on the total weight of the process oil, and has an overall fatty acid content.

In one aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth or twenty-fifth embodiment, the processed oil further comprises oleic acid in an amount from about 5% to about 75% by weight of the total fatty acid content of the processed oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 10% to about 75% by weight of the total fatty acid content of the process oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 20% to about 75% by weight of the total fatty acid content of the process oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 30% to about 75% by weight of the total fatty acid content of the process oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 40% to about 75% by weight of the total fatty acid content of the process oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 50% to about 75% by weight of the total fatty acid content of the process oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises oleic acid in an amount from about 60% to about 75% by weight of the total fatty acid content of the process oil.

In one aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment or any preceding aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the process oil further comprises linoleic acid in an amount from about 1.5% to about 90% by weight of the total fatty acid content of the process oil.

In one aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment or any preceding aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processed oil further comprises linoleic acid in an amount from about 10% to about 25% by weight of the total fatty acid content of the processed oil.

In one aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment or any preceding aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processed oil further comprises linolenic acid in an amount of about 0.1% to about 2% by weight of the total fatty acid content of the processed oil.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linoleic acid, wherein the ratio of oleic acid to linoleic acid is between about 0.01 and 5.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linoleic acid, wherein the ratio of oleic acid to linoleic acid is between about 1 and 4.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linoleic acid, wherein the ratio of oleic acid to linoleic acid is between about 3 and 4.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid is between about 1 and 100.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid is between about 10 and 100.

In another aspect of the twenty-first, twenty-second, twenty-third, twenty-fourth, or twenty-fifth embodiment, the processing oil further comprises oleic acid and linolenic acid, wherein the ratio of oleic acid to linolenic acid is between about 10 and 30.

The foregoing descriptions of specific embodiments of the present disclosure have been presented for purposes of illustration and description. The exemplary embodiments were chosen and described in order to best explain the principles of the disclosure and its practical application, to thereby enable others skilled in the art to best utilize the subject matter and various embodiments with various modifications as are suited to the particular use contemplated. The different features and disclosures of the various embodiments in the present disclosure may be combined within the scope of the present disclosure.

Claims

1. A processing oil comprising Monoacylglycerols (MAG), Diacylglycerols (DAG), and Free Fatty Acids (FFA), wherein MAG comprises about 30 wt% to about 90 wt% of the total weight of the processing oil, wherein DAG comprises about 10 wt% to about 30 wt% of the total weight of the processing oil, wherein FFA comprises about 5 wt% to about 60 wt% of the total weight of the processing oil, and further comprising non-oil components naturally present in the processing oil, and wherein the processing oil is substantially free of Monochloropropanediol (MCPD).

2. The processing oil of claim 1, further comprising Triacylglycerols (TAGs), wherein the TAGs comprise 5 weight percent or less of the total weight of the processing oil.

3. A processing oil according to any of claims 1-2, wherein the non-oil ingredients comprise antioxidants.

4. The processing oil of claim 3, wherein the antioxidant is tocopherol.

5. A process oil according to any one of claims 1-2, wherein the non-oil ingredients comprise vitamins.

6. The processing oil of any one of claims 1-2, wherein the non-oil ingredients comprise one or more of ceramide phosphate, monogalactosyldiacylglycerols, phosphatidylcarbinols, sitosterol esters, campesterol esters, sphingolipids, phosphatidylglycerols, wax esters, and sphingomyelins.

7. The processed oil of any one of claims 1-6, wherein said processed oil is derived from an oil source selected from the group consisting of safflower oil, grape oil, silybum oil, sesame oil, sunflower oil, wheat germ oil, pumpkin seed oil, sesame oil, rice bran oil, almond oil, rapeseed oil, peanut oil, olive oil, and coconut oil.

8. The processed oil of any one of claims 1-6, wherein said processed oil is derived from a combination of two or more of the group consisting of safflower oil, grape oil, silybum oil, sesame oil, sunflower oil, wheat germ oil, pumpkin seed oil, sesame oil, rice bran oil, almond oil, rapeseed oil, peanut oil, olive oil, and coconut oil.

9. The processing oil of any one of claims 1-6, wherein the processing oil is derived from algae or fish.

10. The processing oil of any one of claims 1-9, wherein the processing oil has a fatty acid content comprising oleic acid, linoleic acid, and linolenic acid.

11. The process oil of claim 10, wherein the amount of oleic acid is from 10% to about 75% by weight of the total fatty acid content of the process oil.

12. The processing oil of claim 11, wherein from about 50% to about 99% of the oleic acid content is esterified at the sn-1 position.

13. The process oil of claim 10, wherein linoleic acid is in an amount from 1.5% to about 90% by weight of the total fatty acid content of the process oil.

14. The process oil of claim 10, wherein the amount of linolenic acid is from 0.01% to about 2% by weight of the total fatty acid content of the process oil.

15. A processing oil is prepared by the following processes:

mixing a starting oil with a buffer solution and a first enzyme to produce a first reaction mixture, wherein the starting oil comprises Triacylglycerol (TAG) and Monochloropropanediol (MCPD), wherein the amount of TAG is greater than 50 wt% based on the total weight of the starting oil, wherein the MCPD is present in an amount of MCPD from about 1.00mg/kg to about 12.00mg/kg, and wherein the starting oil is free of Monoacylglycerol (MAG) or is present in an amount less than 5 wt% based on the total weight of the starting oil, wherein the first enzyme is capable of hydrolyzing the TAG to Free Fatty Acids (FFA);

reacting the first reaction mixture under conditions sufficient for the first enzyme to hydrolyze the TAG for a first period of time to produce an aqueous phase and a first lipid reaction product comprising FFA;

inactivating the first enzyme in the first lipid reaction product;

collecting the first lipid reaction product by removing it from the aqueous phase;

mixing the first lipid reaction product with food grade glycerol and a second enzyme capable of esterifying FFA to form a second reaction mixture;

reacting the second reaction mixture for a second period of time to produce a second lipid reaction product comprising a lipid oil phase and a glycerol phase;

inactivating the second enzyme in the second lipid reaction product;

adding a salt to the reaction product and separating the lipid oil phase from the glycerol phase; and

collecting the lipid oil phase, wherein the lipid oil phase is the processing oil, wherein the processing oil comprises MAG, Diacylglycerols (DAG) and Free Fatty Acids (FFA), wherein the amount of MAG is from about 30 wt% to about 90 wt% based on the total weight of the processing oil, wherein the DAG comprises from about 10 wt% to about 30 wt% of the total weight of the processing oil, wherein FFA comprises from about 5 wt% to about 60 wt% of the total weight of the processing oil, and wherein the processing oil is free of TAG or comprises TAG in an amount of from about 0.1 wt% to about 5 wt% based on the total weight of the lipid oil phase, and wherein the lipid oil phase is substantially free of MCPD.

16. The processing oil of claim 15, wherein the starting oil has a fatty acid content comprising oleic acid, linoleic acid, and linolenic acid, and wherein the amount of oleic acid, linoleic acid, and linolenic acid in the processing oil is within 10% of the amount of oleic acid, linoleic acid, and linolenic acid in the starting oil.

17. The processing oil of claim 15 or 16, wherein the starting oil comprises one or more naturally occurring non-oil ingredients selected from the group consisting of ceramide phosphate, monogalactosyldiacylglycerol, phosphatidylmethanol, sitosterol ester, campesterol ester, sphingolipids, phosphatidylglycerol, wax ester, and sphingomyelin, and wherein the one or more naturally occurring non-oil ingredients are retained in the processing oil.

18. A food product comprising a process oil, a carbohydrate source, and a protein source, and having a total weight of at least 25 grams, a caloric density of from about 1 kcal/gram to about 5 kcal/gram, wherein the process oil constitutes from about 5% to about 75% of the total caloric content, wherein the process oil has a Triacylglycerol (TAG) content equal to or less than 5% by weight, based on the total weight of the process oil, wherein the process oil has a Monochloropropanediol (MCPD) of less than 0.10mg/kg, and wherein the process oil has a fatty acid content comprising oleic acid, linoleic acid, and linolenic acid.

19. The food product of claim 18, wherein the processing oil has a Monoacylglycerol (MAG) content equal to or greater than 30 wt% based on the total weight of the processing oil.

20. The food product of claim 18 or 19, wherein the processed oil has a Diacylglycerol (DAG) content of from about 10% to about 30% by weight, based on the total weight of the processed oil.

21. The food product of any one of claims 18-20, wherein the processing oil has a Free Fatty Acid (FFA) content of about 5 wt% to about 60 wt%, based on the total weight of the processing oil.

22. The food product of any one of claims 18-21, wherein the processed oil comprises non-oil ingredients naturally present in the processed oil, wherein the non-oil ingredients comprise one or more of ceramide phosphate, monogalactosyl diacylglycerol, phosphatidyl methanol, sitosterol ester, campesterol ester, sphingolipids, phosphatidyl glycerol, wax ester, and sphingomyelin.

23. The food product of any one of claims 18-22, wherein the amount of oleic acid is from 10% to about 75% by weight of the total fatty acid content of the processed oil.

24. The food product of any one of claims 18-23, wherein about 50% to about 99% of the oleic acid content is esterified at the sn-1 position.

25. The food product of any one of claims 18-24, wherein the amount of linoleic acid is from 1.5% to about 90% by weight of the total fatty acid content of the processed oil.

26. The food product of any one of claims 18-25, wherein the amount of linolenic acid is from 0.01% to about 2% by weight of the total fatty acid content of the processed oil.