Cetenoic acid composition

Abstract

The present invention relates to a composition rich in LCMUFA, which comprises cetenoic acid. Furthermore, the present invention relates to a method for providing an enriched composition comprising cetenoic acid, and to the use of such a composition.

Description

Technical Field

[0001] The present invention relates to LCMUFA compositions, and in particular to LCMUFA-enriched compositions comprising cetoleic acid. Furthermore, the present invention relates to methods for providing compositions comprising cetoleic acid, and to different applications of LCMUFA compositions. Background Art

[0002] A large number of scientific studies support the key role of omega-3 fatty acids in improving risk factors for heart disease, reducing inflammation, and promoting brain health during pregnancy and early life. With such a wide range of proven positive effects on human health, the omega-3 ingredient market is a big business. Looking back to the 1960s, a multinational epidemiological study, Keys et al, Acta Med Scand Suppl. 1966, 460, 1-392 caused such a discovery: long-chain monounsaturated fatty acids (LCMUFA) derived from fish (e.g., mackerel and herring) also have positive health effects, especially those related to coronary heart disease. This study strengthens the general concern about monounsaturated fatty acids and the promotion of the Mediterranean diet rich in oleic acid from olive oil, which is a monounsaturated fatty acid, and the Mediterranean diet is still regarded as one of the healthiest diets.

[0003] The human body contains many different fats, which help us store energy, keep us insulated, protect our vital organs, and provide important structural functions. Several lipids have important biological activities, meaning that they trigger specific responses in the body. The dietary fats consumed by people can be divided into two categories: saturated and unsaturated. The latter is generally regarded as "healthy fat" because it helps to increase the level of good cholesterol HDL and reduce unhealthy triglycerides. Unsaturated fats can be classified into two main types: monounsaturated fat (monounsaturated fat, MUFA) and polyunsaturated fat (polyunsaturated fat, PUFA). Long-chain fatty acids are those fatty acids C20-C22 with carbon chains longer than 18, and fish oil is rich in these lipids. LCPUFA includes ω-3 fatty acids EPA and DHA, which are important for health but are produced in small quantities in people. Many studies have shown the health effects of EPA and DHA, including reducing inflammation, reducing the risk of Alzheimer's disease and reducing the impact of cardiovascular disease. Therefore, the dietary intake of EPA and DHA is important to maintaining the healthy levels of these fatty acids.

[0004] Fish and marine life are rich sources of EPA and DHA, but some fish also contain high levels of LCMUFA. Knowledge about the health benefits of marine LCMUFA is limited compared to EPA and DHA. Cetearyl acid is an omega-11 marine lipid that is found in higher concentrations in North Atlantic fish (such as herring, mackerel, and tobis) compared to South American fish (such as anchovies and sardines) that are richer in EPA and DHA.

[0005] As described by Bremer et al, J Lipid Res, Feb 1982, 23 (2), 43-56, LCMUFAs including cetyl esters can be used as an energy source through mitochondrial β-oxidation and peroxisomal β-oxidation. Adipose tissue is the main storage site for triglycerides, and feeding studies with cetyl ester-rich oils showed high uptake of cetyl esters in adipose tissue, indicating that cetyl esters are stored in adipose tissue and may play an important role in this tissue. Earlier studies, see Yang et al, Lipids Health Dis, Nov 22, 2016, 15 (1), 201 and Yang et al, Mol Nutr Food Res 2016, 60 (10), 2208-2218, have shown that fish oils rich in cetyl esters reduce systemic inflammation and reduce the extent of atherosclerosis in mice. Other positive health effects associated with reducing inflammation and preventing the occurrence of metabolic syndrome and obesity-related metabolic dysfunction have been noted.

[0006] Cell culture studies and human clinical studies support the hypothesis that et al, BR, J Nutr. Oct 14 2019, 122 (7), 755-768, fish oil rich in cetyl esters promotes the conversion of α-linolenic acid to EPA.

[0007] Studies in the late 1990s showed that oils rich in cetyl oleic acid and EPA and DHA may have different activities in vivo. EPA and DHA supplementation leads to beneficial plasma lipid effects, which are not seen in oils rich in cetyl oleic acid. In other words, the health benefits of these two fatty acids can cover different areas of metabolic health. There are several pieces of evidence to support it. In vitro studies using hepatocytes have shown that EPA and DHA reduce the cholesterol and triglyceride content of hepatocytes, but when cells are co-administered with oils rich in cetyl oleic acid, this reduction is greater, see Yoshinaga et al, J Oleo Sci. May 12021, 70 (5), 731-736. Omega-3 fatty acids mainly affect triglycerides and HDL, while oils rich in cetyl oleic acid affect non-HDL cholesterol and insulin sensitivity, with reference to Yang et al, J Agric Food Chem, Jul 13, 2011, 59 (13), 7482-9. This complementary effect aims to provide comprehensive metabolic protection. Indeed, as provided by Yang et al, Lipids Health Dis, Dec 1, 2015, 14, 155, based on data from animal models, a beneficial effect of the combination has been shown in metabolic syndrome, whereby oils rich in EPA / DHA and cetyl esters show both combined and complementary activities within metabolic syndrome, with a combined effect of lowering plasma non-HDL cholesterol, improving hyperinsulinemia, liver fat, and lowering plasma lipid levels.

[0008] As with the LCPUFAs EPA and DHA, levels of ω-11 LCMUFAs such as cetearyl alcohol are dependent on dietary intake since the body cannot synthesize them.

[0009] EP2682116 by Nippon Suisan Kaisha describes an agent for improving metabolic syndrome, wherein the agent contains MUFA but almost no LCPUFA.

[0010] LCMUFA, including cetyl ester, are present in natural oils and are a valuable resource. However, MUFA are often present in a mixture with a range of other fatty acids. Therefore, there is a need for compositions containing LCMUFA, as well as methods for separating MUFA from other fatty acids to provide MUFA compositions. Summary of the invention

[0011] The present invention provides compositions comprising long chain monounsaturated fatty acids (LCMUFA), in particular cetyl ester, methods for providing such compositions, and uses of these compositions.

[0012] In one aspect, the present invention relates to a LCMUFA composition comprising:

[0013] C22:1n11 (cetylene acid) and C20:5n3 (EPA), of which:

[0014] The concentration of C22:1n11 is at least 16.0 wt %; and

[0015] The EPA concentration is at least 3% by weight.

[0016] In the composition of the present invention, the DHA concentration is low. In one embodiment, the DHA concentration is a maximum of 4.0% by weight of the composition. In one embodiment, the EPA:DHA ratio is at least 4:1.

[0017] In one embodiment, the present invention relates to a composition enriched in LCMUFAs, wherein the concentration of C22:1n11 is at least 39% by weight.

[0018] In a second aspect, the present invention relates to a method for preparing a composition of LCMUFAs, said method comprising the steps of:

[0019] i) mixing a natural oil with a lipase to obtain a mixture comprising LCPUFAs in the form of glycerides and LCMUFAs in the form of ethyl esters;

[0020] ii) subjecting the mixture to distillation to obtain a LCMUFA composition as a first fraction and a LCPUFA composition as a second fraction.

[0021] In one embodiment, the method is used to prepare the LCMUFA composition of the first aspect.The method enables LCMUFAs such as cetyl ester and gondoic acid to be separated from EPA and DHA.

[0022] In further aspects, the invention relates to a LCMUFA composition according to the first aspect for use in therapeutic or health indications, and to its use in feed or food. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Results from a study using a wound healing model (Example 4) are presented showing wound area 14 hours after wounding and cell treatment (cells were supplemented with LCMUFA concentrate or DHA, respectively).

[0024] Figure 2 Shown are melanin concentrations in human fibroblasts (μg melanin / μg cellular DNA), n=4, following oxidative stress, wound healing model from Example 4.

[0025] Figure 3The relative expression (log2) of inflammation-related genes in human fibroblasts under oxidative stress, from the wound healing model of Example 4, is shown. DETAILED DESCRIPTION

[0026] Unless otherwise defined, all technical terms, symbols and other scientific terms or terms used herein are intended to have the meanings commonly understood by those skilled in the art to which the present invention belongs. In some cases, for the sake of clarity and / or for timely reference, terms with commonly understood meanings are defined herein; and such definitions contained herein should not necessarily be understood to represent significant differences beyond what is generally understood in the art.

[0027] In the present invention, "monounsaturated fatty acids" or "MUFAs" are fatty acids with one double bond, while "polyunsaturated fatty acids" or "PUFAs" refer to fatty acids with four double bonds or more. MUFAs and PUFAs with long chain fatty acids of 20 or more carbon atoms are described as LCMUFAs or LCPUFAs. MUFAs with 20 carbons are called eicosenoic acid (icosenoic acid) and cis-9-eicosenoic acid (n-11, commonly known as gadoleic acid), cis-11-eicosenoic acid (n-9, commonly known as cis-11-eicosenoic acid), etc. in the IUPAC nomenclature system. MUFAs with 22 carbon atoms are called docosenoic acid, cis-11-docosenoic acid (n-11, commonly known as cetyl enoic acid), cis-13-docosenoic acid (n-9, commonly known as erucic acid), etc. in the IUPAC nomenclature system.

[0028] In the present invention, the following nomenclature is used:

[0029] cis-11-eicosenoic acid, C20:1n9;

[0030] Gedanoic acid, C20:1n11;

[0031] Erucic acid C22:1n9;

[0032] Cetyl ester, C22:1n11;

[0033] Eicosapentaenoic acid (EPA), C20:5n3;

[0034] Docosahexaenoic acid (DHA), C22:6n3.

[0035] The present invention relates to compositions and to methods for providing such compositions, wherein the compositions comprise both LCMUFA and LCPUFA obtained from natural oils. The present invention provides LCMUFA compositions and in particular provides LCMUFA-enriched compositions comprising cetoleic acid. The present invention also provides methods for producing compositions comprising high concentrations of cetoleic acid from natural oils by separating cetoleic acid from other lipids.

[0036] Provided are enriched compositions that provide complementary benefits from EPA and cetyl esters and other fatty acids. Applicants have examined the evidence supporting marine sources of LCMUFA, particularly ω-11 cetyl esters (C22:1n-11) and ω-9 cis-11-eicosenoic acid (C20:1n-9), and provide novel compositions, for example, for the marine lipid ingredient market or for human health.

[0037] Thus, in one aspect, the present inventors provide a LCMUFA composition comprising

[0038] C22:1n11 (cetylene acid) and C20:5n3 (EPA), of which:

[0039] The concentration of C22:1n11 is at least 16 wt %;

[0040] The EPA concentration is at least 3% by weight.

[0041] In the composition of the invention, the DHA concentration is low and therefore the ratio of either EPA:DHA and cetyl ester:DHA is high. In one embodiment, the DHA concentration is a maximum of 4% by weight of the composition.

[0042] The MUFA composition of the present invention containing high concentrations of cetylene acid is derived from a natural starting oil containing at least cetylene acid and EPA. The starting oil, also referred to as raw material, is a crude oil or refined oil used as a starting oil for providing the composition of the present invention, typically containing at least 9% by weight cetylene acid and at least 3% by weight EPA, more preferably at least 15% by weight cetylene acid and at least 5% by weight EPA, for example at least 7% by weight EPA. The applicant has found that it is possible to obtain a composition containing commercially available amounts of LCMUFA and LCPUFA with health value from the same starting oil. A method for separating certain LCMUFA from certain LCPUFA of a separate fraction is disclosed herein. Therefore, the present invention also relates to a method for producing a composition containing high concentrations of LCMUFA, particularly cetylene acid from natural oils, and additionally, such a composition is disclosed and claimed. Such a method has been determined: it enables MUFA C22:1n11 (cetylene acid) to be separated from other fatty acids, including separation from polyunsaturated fatty acids C22:6n3 (DHA) having the same length, as further shown below.

[0043] The raw material for providing the composition of the present invention is a natural oil from a marine biological source, such as from fish oil, squid oil, krill oil or algae oil, and preferably from fish oil. In particular, the raw material is a North Atlantic fish oil, such as oil from herring, mackerel, capelin, cod, saithe or tobis. The raw material may also be from other fish species containing spermaceti, such as pollock or saury, which are North Pacific fish species. Most preferably, the raw material is an oil from herring or mackerel and most preferably from herring. These North Atlantic oils are advantageous due to the higher available volume, due to the high concentration of spermaceti, and due to the EPA:DHA ratio in these oils. In addition, North Atlantic fish oils are rich in cis-11-eicosenoic acid c20:1n9, which is another reason for using, for example, oil to provide the composition of the present invention. Therefore, in one embodiment, the MUFA composition of the present invention provided is a fish oil MUFA composition. In addition, in one embodiment, the fatty acids of the starting oil used to prepare the composition are in the form of triglycerides. The provision of the MUFA composition of the present invention therefore also includes the valuable use of resources in which some materials have been rarely used to date. It is particularly beneficial to be able to preserve and utilize both MUFA and PUFA from MUFA-rich oils. As is the case today, North Atlantic oils are generally considered a second choice for EPA and DHA production because the levels of EPA and DHA are lower than those in oils such as anchovies and sardines. The present invention provides methods in which oils with high MUFA contents are utilized, which provide valuable MUFA compositions and also recover and utilize PUFAs; rather than using conventional EPA / DHA sources such as anchovy oil, or using MUFA-rich oils for EP / DHA production without utilizing the MUFA component.

[0044] In some embodiments, the composition of the present invention comprises 16.0% to 60.0% by weight of cetylene acid, for example 18.0% to 50.0% by weight of cetylene acid, more preferably 20.0% to 50.0% by weight of cetylene acid, even more preferably 39.0% to 50.0% by weight of cetylene acid, and preferably about 40% to 45% by weight of cetylene acid. In one embodiment, the concentration of cetylene acid is at least 39% by weight. The enrichment of cetylene acid can be achieved, for example, as disclosed below.

[0045] In some embodiments, the composition comprises LCMUFA in an amount of at least 20.0% by weight (e.g., at least 30.0%). More preferably, the enriched composition of the present invention comprises LCMUFA in an amount of at least 47% by weight, preferably at least 50% by weight (e.g., at least 60% by weight). In one embodiment, the composition comprises LCMUFA in the range of 47% by weight to 80% by weight (e.g., 50% by weight to 76% by weight), and preferably in the range of 60% by weight to 65% by weight. In addition to spermaceti, LCMUFA also comprises C20 fatty acid cis-11-eicosenoic acid and may further comprise gadoleic acid and C22:1 fatty acid erucic acid. Therefore, the provided composition is rich in LCMUFA (i.e., C20:1 and 22:1), and the concentration of LCMUFA is significantly higher in the claimed composition than in the starting oil used for preparation. However, the ratio of C22:1 to C20:1 is similar to that found in the starting oil (natural oil). In one embodiment, the weight ratio between C22:1 and C20:1 is 1.5:1 to 2.6:1, such as about 1.75: 1. More particularly, the weight ratio of C22:1n11:C20:1n9 is about 1.75: 1. In one embodiment, the weight ratio of C22:1n11:C20:1n9 is 1.5:1 to 2.6:1.

[0046] In some embodiments, the concentration of cis-11-eicosenoic acid C20:1n9 is in the range of 9 wt % to 28 wt %, for example 15 wt % to 28 wt %, for example 18 wt % to 28 wt %. More preferably, the concentration of C20:1n9 is in the range of 18 wt % to 26 wt %. Different from obtaining from typical Pacific fish species such as pollock or saury, such high concentration cis-11-eicosenoic acid combined with high cetyl oleic acid concentration is particularly available from North Atlantic fish oil. Oil from Pacific fish species generally contains more cod oleic acid C20:1n11 rather than cis-11-eicosenoic acid (C20:1n9). Enriched compositions and high concentrations can be obtained by the following disclosed method, particularly when the method comprises more than one concentration round.

[0047] In some embodiments, the amount of gadoleic acid C20:1n11 in the composition of the invention is particularly low. It is a maximum of 2.0% by weight, preferably 1.0% to 2.0% by weight.

[0048] In some embodiments, the amount of erucic acid C22:1n9 in the composition of the invention is particularly low. It is a maximum of 3.0% by weight, preferably 2.0% to 3.0% by weight.

[0049] The EPA concentration in the composition is at least 3.0% by weight, and preferably at least 5.0% by weight. In some embodiments, the concentration is 5.0% to 11.0% by weight. In some embodiments, the EPA concentration is about the same as the concentration in the starting oil (crude oil), or slightly lower. In some exemplary embodiments, for example, in embodiments using mackerel or herring oil as the starting oil, there is an EPA concentration of about 8%, and the composition of the present invention comprises about 5% to 8% EPA. In another embodiment, the EPA concentration is even higher, for example, up to 40% by weight of the composition. Therefore, the present invention provides a composition comprising cetyl oleic acid as disclosed in an enriched amount and an EPA concentration of 5% to 40% by weight (e.g., 20% to 40% by weight). In order to obtain a MUFA composition rich in EPA, an EPA concentrate can be added to the MUFA composition of the present invention.

[0050] Thus, in one embodiment, the EPA-enriched LCMUFA composition of the present invention comprises C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein:

[0051] C22:1n11 has a concentration of at least 16.0 wt %, such as 16 wt % to 60 wt %, such as 30 wt % to 50 wt % of the composition;

[0052] The EPA concentration ranges from 20% to 40% by weight of the composition, for example from 20% to 30% by weight of the composition.

[0053] In one embodiment, DHA may be present in a concentration of a maximum of 4.0% by weight of the composition. In addition, the EPA-enriched MUFA composition preferably also comprises cis-11-eicosenoic acid C20:1n9 in a concentration of 9% to 28% by weight of the composition.

[0054] In some preferred embodiments, the compositions of the present invention contain only small amounts of DHA. The applicant was unexpectedly able to separate LCMUFA, particularly C22:1 MUFA, namely cetyl and erucic acid, from the polyunsaturated C22 fatty acids. Thus, the MUFA compositions of the present invention provided are significantly lower in DHA than in the starting oil, and therefore have a high ratio of EPA:DHA. The MUFA compositions of the present invention typically have a cetyl:DHA ratio that is about 10 times higher than the natural starting oil. Crude oil of mackerel typically contains about 11% to 12% DHA by weight, while herring oil typically contains about 9% to 10% DHA by weight. However, the MUFA compositions of the present invention contain very little DHA, but have a high concentration of cetyl, a fatty acid of the same length. In one embodiment, the ratio of cetyl:DHA of the LCMUFA composition of the present invention is at least 10:1, for example at least 15:1, more preferably at least 20:1.

[0055] In addition, the MUFA compositions of the present invention have a high ratio of EPA:DHA. In one embodiment, the LCMUFA composition has an EPA:DHA ratio of 4.0:1 to 16.0:1, more preferably 4.0:1 to 8.0:1. In compositions of the present invention in which the amount of EPA is high, such as in EP-enriched MUFA compositions, the DHA content is low anyway, and the EPA:DHA ratio can be as high as 50:1. Thus, in one embodiment, the EPA:DHA ratio is 4:1 to 50:1. Thus, the concentration of EPA is at least four times higher than the concentration of DHA in the MUFA composition. However, the more the composition is enriched in cetyl esters (e.g., by concentration by distillation), the lower the ratio.

[0056] Therefore, the concentration of DHA is unexpectedly low in the MUFA composition provided. In some embodiments, the DHA concentration of the MUFA composition is a maximum of 4.0% by weight, for example, in the range of 0.5% to 3.5% by weight, i.e., it is significantly lower than that in suitable marine bio-crude oil. Compositions in which there is no DHA, i.e., compositions in which the DHA concentration is 0% by weight, fall within the claimed composition, i.e., compositions in which the DHA concentration is 0.0% to 4.0% by weight. A small amount of DHA may be desirable in the composition, for example, for cardiovascular health, such as for the treatment of heart disorders to avoid negative effects on blood parameters such as TG, LDL, HDL and total cholesterol. In addition, since the composition of the present invention comprises a high concentration of MUFA and very little DHAPUFA, the fatty acids of the composition of the present invention are less susceptible to oxidation than compositions with a higher concentration of PUFA. Therefore, the composition of the present invention has a pleasant taste and smell and can be included in food.

[0057] A further reason for providing the claimed composition is that the applicant wishes to provide a product with cetylene acid and a small amount of DHA and EPA to give consumers a choice so that they can manage their lipid intake. Consumers may already be taking omega-3 supplements, and from a commercial perspective, it is undesirable to replace conventional highly concentrated omega-3s that have a dominant position in the fish oil market. In addition, alternative oils such as oils rich in cetylene acid will never supply the levels of EPA and DHA sold in omega-3 concentrates. This leaves such a problem: consumers can take one omega-3 capsule and one cetylene acid capsule, but have the risk of doubling their EPA and DHA intake, which is regulated by the FDA, with a maximum EPA and DHA intake of 3g. Therefore, consumers have the risk of taking too much omega-3 when taking additional cetylene acid supplements. Therefore, low EPA-DHA cetylene acid supplements are needed. In addition, it is known that EPA and DHA reduce the plasma level of triglycerides (TG) and have a small change in total cholesterol. EPA causes a slight decrease in total cholesterol levels, while DHA shows a slight increase mainly due to an increase in HDL cholesterol. As shown by Sarbolouki et al, 2013, Singapore Med J., 54 (7): 387-90, EPA supplementation can improve insulin sensitivity by reducing fasting plasma glucose, HbA1c and homeostatic model assessment of insulin resistance. Therefore, EPA shows a negative correlation with plasma levels of TG and insulin. However, EPA shows only a slight effect to no effect on total cholesterol. The main claim of LCMUFA cardiovascular protection comes from its ability to reduce LDL levels without reducing HDL levels. Long-term supplementation with LCMUFA (Yang, 2015) showed that LCMUFA had a slight effect to no effect on reducing plasma TG, but significantly reduced plasma levels of insulin and total cholesterol. Therefore, LCMUFA shows a negative correlation with plasma levels of total cholesterol and insulin. However, LCMUFA is neutral in its ability to reduce plasma triglycerides.

[0058] Therefore, LCMUFA and EPA appear to have complementary effects on plasma lipids and insulin. Therefore, the positive effects of LCMUFA (cetylene acid) support the hypothesis that LCMUFA may have a beneficial effect in reducing the risk of cardiovascular disease. The intake of LCMUFA may also have a positive effect on the treatment of diabetes-2 by reducing non-HDL cholesterol and plasma insulin. Many people believe that cardiovascular health from omega-3 is due to the effect of EPA. The success of the drug icosapent ethyl (Vascepa) (REDUCE-IT trial), which contains 99% EPA, and Epadel (Jellis trial), which contains more than 98% EPA, shows this. One object of the present invention is to provide a high concentration of cetylene acid with low levels of EPA and DHA to allow consumers to easily make an informed choice. However, the product favors EPA rather than DHA because EPA is considered to be more important for cardiovascular health.

[0059] Due to the complementary effects of LCMUFA and EPA, the applicant is also interested in providing products for clinical purposes that contain enriched cetylene acid and EPA. Small amounts of DHA would provide the option of preparing such a product by mixing a cetylene acid concentrate with an EPA concentrate. However, if the LCMUFA fraction is already enriched in DHA, the production of such a composition is more challenging. The disclosed method effectively separates cetylene acid from DHA, which significantly increases the concentration of cetylene acid, optionally retaining low levels of DHA.

[0060] In some embodiments, the combined concentration of EPA and DHA is in the range of 3.0% to 12.0% by weight of the composition, e.g., 5.0% to 12.0% by weight, more preferably 6.0% to 12.0% by weight. In some embodiments in which the composition is enriched with EPA, the combined EPA and DHA concentration may be significantly higher.

[0061] In addition to cetylene acid and EPA and preferably cis-11-eicosenoic acid, the MUFA compositions of the present invention may also contain a variety of other fatty acids, including short-chain and medium-chain (C18 or less), long-chain (C20 to 22) or even very long-chain fatty acids (above C22), which are typically found in the crude oil from which the MUFA composition is derived. These fatty acids can be saturated, monounsaturated or polyunsaturated. In some embodiments, the MUFA composition contains short-chain fatty acids. However, naturally, the more concentrated the MUFA composition is in cetylene acid, the lower the concentration of other fatty acids, including shorter fatty acids, will be. Preferably, short-chain saturated fatty acids are kept low, especially in the highly cetylene acid-enriched compositions of the present invention. In some embodiments, the concentration of C12:1 to C16:1 fatty acids does not exceed a maximum of 2.0%. In some embodiments, the MUFA composition contains C18 fatty acids, such as:

[0062] C18:0, e.g., in the range of 0.5 wt % to 2.7 wt %;

[0063] C18: 1, e.g., in the range of 5.0 wt % to 16.0 wt %;

[0064] C18:2, e.g., in the range of 0.5 wt % to 2.5 wt %;

[0065] C18:3, for example in the range of 0.5 wt % to 2 wt %.

[0066] C18:4, for example in the range of 0.5 wt % to 1.5 wt %.

[0067] In an exemplary embodiment, the LCMUFA-enriched composition of the present invention comprises

[0068] C22:1n11 (cetosyl ester) and C20:5n3 (EPA), among which;

[0069] The concentration of C22:1n11 is at least 39% by weight.

[0070] The concentration of EPA is at least 5.0 wt%.

[0071] Furthermore, in some embodiments, the MUFA-rich composition of the present invention has one or more of the following characteristics:

[0072] The concentration of C22:1n11 is at least 39 wt %;

[0073] The weight ratio of C22:1n11:C20:1n9 is 1.5:1 to 2.6:1.

[0074] The concentration of EPA is at least 5.0% by weight;

[0075] The concentration of DHA does not exceed a maximum of 4.0% by weight;

[0076] The concentration of C20:1n11 (codoleic acid) does not exceed a maximum of 2.0% by weight;

[0077] The concentration of C12:1 to C16:1 fatty acids does not exceed a maximum of 2.0%.In a preferred embodiment, the MUFA-enriched composition meets all of the above characteristics.

[0078] In the applicant's analysis, gas chromatography has been used, and the GC area % has provided the information about the fatty acid distribution of the analyzed composition.The fatty acid composition provided is preferably composed of a lipid acid approaching 100%, and therefore only has negligible differences between the name " GC area % " that uses in analysis and the embodiment and " % weight " (weight %) that uses in whole this specification and the claims.

[0079] In certain embodiments, the MUFA-enriched compositions of the present invention comprise the following fatty acids within the ranges as listed in Table A.

[0080] Table A:

[0081]

[0082]

[0083] In another aspect, the invention provides a method for preparing a composition of C20-22 LCMUFA, for example as according to the first aspect, wherein the method comprises the step of selectively separating LCPUFA from LC-MUFA enzymatically. The method comprises the step of selective transesterification by using a lipase. In one embodiment, the method comprises the step of separating LCPUFA from LCMUFA by selectively cleaving LCMUFA from triglycerides enzymatically.

[0084] The method of the present invention allows MUFA and PUFA of the same length to be separated from each other under mild conditions. More particularly, MUFA C22:1n11 (cetylene acid) is separated from PUFA C22:6n3 (DHA). Therefore, the method enables fatty acids with similar boiling points and therefore difficult to separate to be selectively separated from each other in practice. Other methods may enable C20 fatty acids to be separated from C22 fatty acids, but if C22:1 is concentrated from C20 fatty acids, for example, DHA (C22:6) will usually be together with C22:1 fatty acids.

[0085] Therefore, in one embodiment of the present invention, there is provided a method for preparing a composition of LCMUFAs, the method comprising the following steps:

[0086] i) mixing a natural oil, i.e. a natural oil comprising LCMUFAs and LCPUFAs, with a lipase to obtain a mixture comprising LCPUFAs in the form of glycerides and LCMUFAs in the form of ethyl esters;

[0087] ii) distilling the mixture of step i) to obtain a LCMUFA composition as one fraction and a LCPUFA composition as another fraction.

[0088] Furthermore, the present invention provides a method for separating LCMUFA from LCPUFA, comprising the following steps:

[0089] i) mixing a natural oil with a lipase to obtain a mixture comprising LCPUFAs in the form of glycerides and LCMUFAs in the form of ethyl esters;

[0090] ii) subjecting the mixture of step i) to distillation to obtain a LCMUFA composition as a first fraction and a LCPUFA composition as a second fraction.

[0091] In these methods, the enzymatic reaction occurs on triglycerides, which is beneficial. Since DHA fatty acids tend to be located in the middle of triglycerides, fatty acids located in 1 and 3 (such as LCMUFA) can be selectively cut off by using 1,3-specific lipases. DHA will therefore remain on glycerol as DHA monoglyceride (MAG-DHA). Therefore, both regioselectivity and fatty acid selectivity are achieved.

[0092] In an alternative embodiment, further comprising transesterification using a lipase, the present invention provides a method for separating LCMUFA from LCPUFA comprising the steps of:

[0093] Converting fatty acids of natural oils into ethyl esters;

[0094] distilling the ethyl esters to obtain a fraction of LCMUFA and LCPUFA in the form of ethyl esters;

[0095] The ethyl esters are mixed with glycerol and lipase to obtain a glyceride mixture containing LCMUFA. The DHA will remain as the ethyl ester.

[0096] In the method of the present invention, as disclosed above for the composition of the present invention, the starting material is a natural oil comprising at least cetyl oleic acid (LCMUFA) and EPA (LCPUFA). The starting oil (also referred to as raw material) is a crude oil or refined oil used as the starting oil for providing the composition of the present invention, typically comprising at least 9 wt % cetyl oleic acid and at least 3 wt % EPA, more preferably at least 15 wt % cetyl oleic acid and at least 5 wt % EPA, for example at least 7 wt % EPA. The natural oil used for the method can therefore be a crude oil, such as the crude oil typically produced in fish meal production. Alternatively, the natural oil can be a refined oil, such as an oil from which pollutants have been removed, such as it has undergone bleaching, deacidification, stripping or supercritical extraction. The natural oil is from a marine biological source, such as from fish oil, squid oil, krill oil or algae oil, and preferably from fish oil. In particular, the oil is a North Atlantic fish oil, such as from the oil of herring, mackerel, capelin or tobis. Most preferably, the oil is the oil from herring or mackerel, and most preferably from herring. The fatty acids of the starting material are preferably in the form of triglycerides.

[0097] In step i) of the method, enzymatic separation is performed by using a lipase. Lipase is a family of enzymes that catalyze the hydrolysis of fats. Some lipases show a wide range of substrates, including esters of cholesterol, phospholipids and fat-soluble vitamins. In the method of the present invention, at least one lipase is used as an esterification catalyst to cut fatty acid chains from the triglyceride backbone. As known, lipases are very suitable for use as catalysts in processes involving highly unstable n-3 polyunsaturated fatty acids (such as EPA and DHA) present in marine oils. This is due to their ability to work at low temperatures, their neutral pH and their gentle action, which helps to keep undesirable side reactions such as cis-trans isomerization, double bond migration, polymerization, oxidation, etc. to a minimum. Therefore, the use of lipases to hydrolyze fatty acids in marine oils has been well documented. However, the applicant unexpectedly found that certain lipases can be selectively used to separate MUFA from PUFA.

[0098] In step i), lipase is contacted with the fatty acid of natural oil, thereby LCMUFA is converted into LCMUFA ester. In the case where LCMUFA is converted into ester from triglyceride form, LCPUFA will remain as glyceride. Thus, a mixture of LCPUFA in the form of glyceride and LCMUFA in the form of ethyl ester is obtained. The PUFA of this mixture will generally comprise a mixture of monoglyceride, diglyceride and triglyceride. In one embodiment, ethanol is used as substrate and the MUFA of the mixture of step i) will be in the form of MUFA ethyl ester. Solvent is usually not required, but can be used in some cases (for example, if the oil is extremely rich in stearin).

[0099] Reaction conditions, including temperature, pressure and reaction time, are selected based on the normal operating conditions used when converting triglycerides into ethyl esters by using the same enzyme. The reaction with lipase is typically run over a period of time, such as 1 to 48 hours, such as 20 to 30 hours, such as about 24 hours. Typically, a temperature in the range of 25° C. to 90° C. is suitable, and a pressure of 1 to 50 mbar is suitable, if desired.

[0100] In most cases a suitable amount of lipase will be about 10% (w / w) or less of the oil.

[0101] The method of the present invention utilizes a lipase active in catalyzing the esterification of MUFA from triglycerides. Suitable lipases are preferably immobilized enzymes, but non-immobilized enzymes may also work, but recovery after use is foreseen to be more difficult. In one embodiment, the lipase is a 1,3-specific lipase. Some examples of lipases that can be used are Rhizomucor miehei lipase (formerly known as Mucor miehei lipase), Aspergillus niger lipase, Thermomyces lanuginosus lipase, Candida antarctica lipase, Candida rugosa lipase (formerly known as Candida cylindracea lipase), Geotrichum candidum lipase, Penicillium roguefortii lipase, Rhizopus delemar lipase, and Rhizopus oryzae lipase. More preferably, the lipase is selected from the group consisting of Rhizomucor miehei lipase, Thermomyces lanuginosus lipase, Candida antarctica lipase, and Candida rugosa lipase.

[0102] When the reaction of step i) is complete, the material may be cooled and filtered prior to step ii).

[0103] In step ii), the mixture of step i) is subjected to distillation to separate the LCPUFA glycerides of the mixture of step i) from the LCMUFA ethyl esters of the mixture of step i), obtaining a LCMUFA composition as a first fraction and a LCPUFA composition as a second fraction. In one embodiment, the distillation is a single column distillation, such as a single column short path distillation.

[0104] The obtained LCMUFA composition has a higher concentration of LCMUFA, in particular cetyl esters, than in the natural starting oil. In addition, the obtained LCMUFA composition has only a very small amount of DHA present, because this C22 PUFA has been separated from the C22 MUFA and is present in the LCPUFA fraction (PUFA stream). Likewise, the LCPUFA composition has a higher concentration of LCPUFA than in the starting oil.

[0105] The method of the present invention allows MUFA and PUFA of the same length to be separated from each other under mild conditions. More particularly, MUFA C22:1n11 (cetylene acid) is separated from PUFA C22:6n3 (DHA). The first fraction rich in cetylene acid obtained from the method can be further concentrated to prepare an even richer composition of MUFA, or can be used as is. The separated EPA-DHA-rich fraction (PUFA stream) can also be recovered, for example for commercial use. Therefore, in another aspect of the present invention, a method for producing a MUFA composition according to the first aspect is provided.

[0106] To illustrate what can be achieved by this method, as an example, starting from mackerel oil containing about 16% by weight of cetylene acid, 8% EPA and 12% DHA, a LCMUFA composition containing about 21% by weight of cetylene acid can be obtained by the method of the present invention without any further enrichment. In such a LCMUFA stream obtained from enzymatic separation and primary distillation (steps i and ii), the EPA concentration obtained is generally about 6%, while the DHA concentration is only 1.3%. Therefore, by using the method of the present invention, the cetylene acid concentration of the LCMUFA composition is significantly increased, while the DHA concentration is significantly reduced. The applicant has found that when using the method of the present invention, the average change in the concentration of cetylene acid from crude oil is about 2%. The present invention therefore provides a method for producing a composition of C20-22 LC-MUFA. The MUFA composition obtained can be as disclosed in the first aspect, comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein:

[0107] The concentration of C22:1n11 is at least 16 wt %; and

[0108] The EPA concentration is at least 3% by weight.

[0109] In one embodiment, the DHA concentration in the obtained composition is a maximum of 4.0% by weight of the composition. In one embodiment, the EPA:DHA ratio is at least 4.0. Further embodiments of the obtained composition are as disclosed in the first aspect.

[0110] Equivalently, the method of the invention for separating LCMUFA from LCPUFA also provides a method for producing an enriched composition of LCPUFA. Starting from North Atlantic fish oil (e.g., such as mackerel oil), the PUFA stream from the disclosed method (steps i and ii) can typically contain about 9 wt % to 12 wt % EPA and 13 % to 15 % DHA. Thus, the method provides an efficient procedure for separating MUFA from PUFA and providing such a more concentrated composition.

[0111] The LCMUFA composition obtained from the enzymatic separation and primary distillation ("LCMUFA stream") is found to be valuable, for example, as an energy source for food, feed, or as a source for providing a composition more highly enriched in LCMUFA. In one embodiment, the LCMUFA composition is used in aquaculture, for example as part of fish feed. Further applications of the LCMUFA composition of the present invention are disclosed below, also including use in therapeutic or health indications.

[0112] The LCPUFA composition obtained from the enzymatic separation and primary distillation ("LCPUFA stream") is found to be valuable, for example, as an energy source for feed or as a source for providing a composition more highly enriched in LCPUFAs. This may be a valuable alternative source of EPA and DHA to anchovy oil from sustainable fish resources. These EPA and DHA can therefore be further processed into different EPA and DHA concentrates and used, for example, in nutritional products or pharmaceuticals.

[0113] The LCMUFA composition and the LCPUFA composition obtained from step ii) can be further concentrated to form a more enriched composition, respectively. Therefore, in one embodiment, the LCMUFA first fraction composition (from step ii) is further concentrated to provide a more concentrated LCMUFA composition, i.e., a more enriched composition containing a higher concentration of cetylene acid. As an example, the separated mackerel oil containing about 21% by weight of cetylene acid disclosed above can be further concentrated after steps i) and ii) to increase the amount of cetylene acid. As disclosed in the first aspect, a composition containing at least 39.0% by weight of cetylene acid can be obtained accordingly. As shown in Examples 2 and 3, a composition containing about 43% to 45% by weight of cetylene acid was obtained from mackerel or herring oil. Such further concentration to form an enriched composition of LCMUFA can be achieved using methods such as distillation, extraction, enzymatic processing, chromatography and / or other fractionation methods known to those skilled in the art. Such further concentration is preferably accomplished by one or more distillations, such as a high-quality molecular / short-path distillation procedure. The method thus optionally further comprises a step iii) of concentrating the desired PUFA or MUFA of the first and second fractions, respectively. Such concentration can be achieved using methods such as distillation, extraction, enzymatic processing, chromatography, and is preferably achieved by distillation such as molecular distillation (short-path distillation). In one embodiment, the method accordingly comprises an optional additional step of concentrating LCMUFA, in particular cetyl ester, for example by distillation. Such an optional additional distillation is thus a step for preparing a higher concentration of cetyl ester and still containing a small amount of DHA.

[0114] In one embodiment, step iii) comprises one or more rounds of distillation, and the fraction containing the most cetylene acid in each round is distilled again. By such a method, a composition comprising at least 39% by weight of cetylene acid, such as 39.0% to 60.0% by weight of cetylene acid, as disclosed in the first aspect, can be obtained. By the method of the present invention, for example comprising several distillations after the enzymatic separation, cetylene acid can be further purified and recovered at a concentration also greater than 50.0% by weight, such as greater than 60.0% by weight, greater than 70.0% by weight, or even greater than 80% by weight.

[0115] In one embodiment, the present invention provides a method for producing an enrichment composition of C20-22 LC-MUFA, the enrichment composition comprising

[0116] C22:1n11 (cetylene acid) and C20:5n3 (EPA), of which:

[0117] The concentration of C22:1n11 is at least 39 wt %;

[0118] The concentration of EPA is at least 5.0 wt.

[0119] By using the enzymatic method of the present invention, it was unexpectedly found that compositions having high concentrations of cetanoic acid, the desired range between cetanoic acid and cis-11-eicosenoic acid, and the desired EPA concentration can be prepared. More particularly, the method enables the provision of compositions in which the DHA concentration is low, and which are separated from MUFAs of the same length. In particular, the provision of compositions comprising: high concentrations of 20:1 and 22:1 MUFAs, and concentrations greater than 5% EPA combined with very low DHA concentrations (e.g., in the range of 0.5% to 3.5%) is achieved. Other methods, such as distillation alone, or even chromatography, cannot achieve the preparation of such a combination (as shown by the following examples).

[0120] The method of preparation may also include a step of removing or reducing any toxic components. The purified and upwardly concentrated compositions of the present invention also have very small amounts of undesirable contaminants. For example, the amount of oligomeric and polymeric byproducts (including oxidation products) is reduced from their amount in the starting oil. In addition, since the DHA level is low in the composition, the tendency to oxidation is low, which is beneficial to the stability of the composition. Preferably, such oxidation products are a maximum of 1.5% by weight of the fatty acid composition, for example a maximum of 1.0% by weight, more preferably a maximum of 0.5% by weight. More particularly, the amount of environmental pollutants such as benzo(a)pyrene (BAP) and polyaromatic hydrocarbons (PAH) is low in the compositions of the present invention. In one embodiment, the fatty acid mixture of the composition contains less than 2 μg / kg of benzo(a)pyrene (BAP). In another embodiment, the composition preferably contains less than 10 μg / kg of polyaromatic hydrocarbons (4PAH). 4PAH is defined as benzo(a)anthracene, The sum of benzo(b)fluoranthene and benzo(a)pyrene. To measure oxidation in oils there are two main methods in use, to measure the peroxide value and the p-anisidine value.

[0121] Both the fatty acids MUFA and PUFA of the composition and the other fatty acids of the composition may be in different forms. In one embodiment, the fatty acids of the composition are in a form selected from the following alone or in combination: free fatty acids; fatty acid salts; monoglycerides, diglycerides, triglycerides; esters, such as ethyl esters; wax esters; O-acetylated ω-hydroxy fatty acids (OAHFAs); cholesterol esters; ceramides; phospholipids and sphingomyelins. Alternatively, the fatty acids may be in any form that can be absorbed in the digestive tract or absorbed by the body surface after topical application. Preferably, the fatty acids are in the form of free fatty acids, fatty acid salts, ethyl esters or glycerides. In one embodiment, LCMUFA and LCPUFA are independently selected from the following: free fatty acids, fatty acid esters and monoglycerides, diglycerides or triglycerides. In a preferred embodiment, the MUFA of the MUFA composition of the present invention is in the form of ethyl esters. If desired, the form of the fatty acid can be converted to other forms. The skilled person will, for example, know how to transfer, for example, ethyl esters to free fatty acids or glycerides, and the production method may include such steps.

[0122] When mentioning the weight % of lipid acid in the mixture, any above-mentioned most widely limited lipid acid form can be used as the basis for calculating.In addition, the lipid acid of the composition provided in any form listed above is preferably not connected with other active components.Therefore, the fatty acid mixture of composition is pure, unreacted, highly concentrated MUFA and PUFA mixture.But the fatty acid end group can be from original (for example as from glyceride) to ester or free fatty acid or on the contrary modified.

[0123] The LCMUFA compositions of the present invention can be used in a range of different applications.

[0124] The LCMUFA composition disclosed according to the first aspect is found to be valuable, for example, as an energy source for food, supplement, feed, treatment or as a source for providing a composition more highly enriched in LCMUFA. In particular, the MUFA composition of the invention which is a first fraction comprising, for example, about 10% to 25% cetylene acid from enzymatic separation and primary distillation may have commercial purposes, for example, for use in the food or feed industry.

[0125] In one aspect, the present invention provides the use of a MUFA composition as disclosed in the first aspect in food or feed, the MUFA composition comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein:

[0126] The concentration of C22:1n11 is at least 16 wt %;

[0127] The EPA concentration is at least 3% by weight.

[0128] Other embodiments of the composition are as described in the first aspect.

[0129] In an exemplary embodiment, the MUFA composition of the present invention is included in a feed or food composition, which will typically contain a certain amount of unsaturated fatty acids. Since the claimed composition has high stability and is not prone to oxidation, the claimed MUFA composition can beneficially replace the more easily oxidized oils used in food or feed. Another argument for using the MUFA composition in food, feed, or in supplements or nutraceuticals is that cetyl esters can act as a catalyst for the conversion of ALA to EPA and DHA.

[0130] In another exemplary embodiment, the MUFA composition of the invention is included in a feed, such as for aquaculture, such as, in particular, for fish feed compositions, such as feed pellets for fish farming. In particular, the composition is used in a therapeutic feed for fish, such as farmed salmon. In one example, this is used for preventive or therapeutic treatment of heart health.

[0131] In some embodiments, the present invention relates to methods and compositions for treating and alleviating disease or for health indications. As discussed in the background section, studies have been conducted, particularly those by Yang et al, supporting that cetyl esters, for example, in combination with EPA, have beneficial health effects. The MUFA-rich compositions of the present invention comprising high concentrations of cetyl esters as disclosed in the first aspect may have one or more of the following health benefits: providing: reduced inflammation, improved TG and cholesterol, increased fat burning, reduced fatty liver, improved insulin sensitivity, improved metabolic syndrome. As shown in Example 6, the compositions of the present invention can be used to positively affect metabolic syndrome. It can affect parameters related to: diabetes (glucose tolerance, insulin sensitivity, glucose uptake in muscle), diabetic neuropathy - especially allodynia (temperature sensitivity) and metabolic syndrome (liver fat content) and cardiovascular health (blood pressure).

[0132] In some embodiments, the composition is a nutraceutical, supplemental, or pharmaceutical composition, and the composition is used to treat, e.g., prevent, reduce, and / or alleviate the effects, symptoms, etc., of at least one health problem in a subject in need thereof. The MUFA composition is administered to a subject. In one embodiment, the composition is used to maintain good health, e.g., maintain a healthy cardiovascular system / heart health. In at least some embodiments of the invention, the composition does not include an additional active agent. In this embodiment, the composition can be used for medical treatment of a subject, e.g., a subject diagnosed with reduced MUFA levels.

[0133] In one embodiment, the composition according to the present invention is used for human health, for example, in the form of a supplement, i.e., a food supplement, a nutritional supplement or a dietary supplement; a drug, a medical food; or a special medical formula. In the latter group, the composition can be selected from the following: enteral formula for special medical use, food for specified health use, food for special medical purpose (FSMP), food for special dietary use (FSDU), medical nutrition and medical food. Such a composition is particularly suitable for subjects who lack certain nutrients such as cetyl ene acid. The composition is suitable for the nutritional management of subjects with unique nutritional needs. Such a composition is usually administered to the subject under medical supervision. The composition contains relevant MUFA to increase or correct the level of MUFA in the blood or specific tissues of subjects diagnosed with reduced ability to synthesize MUFA and / or low ω-3 index, for example. The composition and method of the present invention have the ability to correct nutritional deficiencies in such a target population. In an exemplary use, the composition is used to maintain a healthy cardiovascular system / heart health. In particular, the composition of the present invention is useful as a dietary supplement for cardiovascular health.

[0134] Applicants are also exploring the potential beneficial effects of the MUFA compositions of the present invention in skin health, which appears to be an important area for the future. Under conditions where the skin lacks the ability to produce MUFAs, the skin is characterized by poor development of the sebaceous glands, which normally keep the skin from drying out. This provides an indication of the importance of MUFAs in skin biology. In one embodiment, the composition is used to treat the skin, such as to aid in the barrier function of the skin, to keep the skin appearing healthy, to avoid skin wrinkles or erythema, and also to protect the skin from the negative effects of sunlight UV radiation. Diseases and conditions related to the skin and hair that can be treated by the composition used in the present invention include at least the following: dry skin and wrinkles, irritation, sourness or sensitivity of the skin, the ability to heal wounds, protection (i.e., preventive treatment) of the skin from the negative effects of sunlight UV radiation, negative effects on hair follicles, and reduced hair health (including the risk of hair loss). Some examples of skin diseases and conditions that typically give skin irritation / sourness and that can benefit from treatment with the composition used are, for example, eczema, psoriasis, dermatitis, acne, and rosacea (papulopustular rosacea). In one embodiment, the MUFA composition of the invention is included in a formulation for use on the skin, such as a cosmetic product. By using the composition or method of the invention, the fatty acid composition of tissue (e.g., skin) can be normalized, for example by compensating for an abnormal sebum fatty acid composition. Such uses or methods may be considered therapeutic or non-therapeutic.

[0135] Reference is made to Examples 4 and 5 below. The effects of the compositions of the present invention on the skin, such as the effects on human fibroblasts, can be viewed as improving the ability of cells to regenerate (wound healing) after injury to affect the production of melanin (skin pigment), or as reducing the expression of genes involved in inflammation (Example 4). In one embodiment, the use of the composition results in an improvement in the grade and severity of eczema (e.g., atopic dermatitis) (Example 5). The use of the composition can show changes in molecular markers such as lipid mediators, inflammatory markers, and blood lipids. More particularly, the use of the composition can positively affect one or more of the following: ceramide / lipid composition in the skin, inflammatory signaling molecules in the skin, ω-3 index in erythrocytes, cetyl ester in erythrocytes, cholesterol / lipid profile, trans-epithelial water loss (TEWL), hydroxylated ω-3 fatty acids in plasma, correlation of ω-3 index with eczema area and severity index (EASI) parameters, correlation of cetyl ester content in erythrocytes with EASI parameters.

[0136] Thus, in one embodiment, the present invention provides a composition of the first aspect for use as a nutraceutical, a food supplement, a food additive or a cosmetic product.

[0137] Compositions according to the present invention, such as supplements, can be delivered or administered in any suitable form, including but not limited to oral delivery, transdermal delivery or transmucosal delivery, including as eye drops. The composition of the present invention can be formulated with other ingredients to include acceptable excipients and / or carriers for oral use, and in particular in the form of an oral delivery vehicle, such as a capsule, preferably a gelatin capsule, a liquid, an emulsion, a tablet or a powder.

[0138] Thus, the present invention provides a MUFA composition as disclosed in the first aspect for use in treatment or health indications, comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein:

[0139] The concentration of C22:1n11 is at least 16 wt %;

[0140] The EPA concentration is at least 3% by weight.

[0141] Likewise, the present invention provides a method of treatment, which administers to a subject a MUFA composition as disclosed in the first aspect, the MUFA composition comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein:

[0142] The concentration of C22:1n11 is at least 16 wt %;

[0143] The EPA concentration is at least 3% by weight.

[0144] In some embodiments, the composition used is a LCMUFA-enriched composition comprising:

[0145] C22:1n11 (cetylene acid) and C20:5n3 (EPA), of which:

[0146] The concentration of C22:1n11 is at least 39 wt %;

[0147] The concentration of EPA is at least 5.0 wt.

[0148] Thus, the present invention provides a composition according to the first aspect for use as a supplement, a medicament, a medical food or as a special medical use formula, or for application to the skin.

[0149] The MUFA composition used is as described in detail in the first aspect. In particular, the composition for treatment or for the method of treatment is used to reduce inflammation, improve TG and cholesterol, increase fat burning, reduce fatty liver, or improve insulin sensitivity or improve metabolic syndrome, which are indications in which cetyl oleic acid and EPA have combined benefits. In addition, the composition of the present invention can be used for one or more of the following: reducing triacylglycerides, increasing HDL cholesterol, lowering blood pressure, reducing inflammation, and as a precursor of a specific proresolving mediator (SPM). In addition, the composition of the present invention is used for one or more of the following: reducing non-HDL cholesterol, reducing hyperinsulinemia, glucose regulation, and reducing inflammation, which are all the effects of cetyl oleic acid. With reference to the research of Yang et all, 2016, the development of the MUFA composition of the present invention with specific therapeutic benefits has been considered. First, reduce the risk of cardiovascular disease; and second, as an alternative means of providing ω-3 levels in humans, thereby providing benefits in areas such as chronic inflammation, cognition, and healthy aging. et al, BR, J Nutr. Oct 14, 2019, 122(7), 755-768, cell culture studies and human clinical studies support the hypothesis that fish oil rich in cetyl oleic acid promotes the conversion of α-linolenic acid into EPA, which supports the use of cetyl oleic acid-rich oil as a relevant source of EPA and DHA as the claimed composition. In one embodiment, the composition is used to treat one or more of the following: metabolic syndrome, fatty liver disease (NAFLD, NASH), diabetes, prediabetes, and cardiovascular disease.

[0150] In another aspect, the present invention provides a combined formulation. Such a formulation may be a combination of a MUFA composition as disclosed in the first aspect with an additional marine oil. As an alternative, the MUFA composition of the present invention is combined with anchovy oil or a fraction from anchovy oil, for example to further concentrate the composition with PUFAs, thereby providing a formulation that can be used for different types of applications.

[0151] The present invention should not be limited to the embodiments and examples shown. Although multiple embodiments of the present disclosure are described herein, it will be apparent to those skilled in the art that such embodiments are provided only by way of example. Without departing from the present disclosure, multiple modifications and changes and variations and replacements of the embodiments described herein will be apparent to those skilled in the art. It should be understood that, in practicing the present disclosure, multiple alternatives to the embodiments described herein may be adopted.

[0152] It should be understood that each embodiment of the present disclosure may be optionally combined with any one or more of the other embodiments described herein.

[0153] It should be understood that embodiments disclosed for one aspect are also applicable to other aspects of the invention. For example, embodiments disclosed for compositions are also applicable to aspects involving methods of production and uses of compositions.

[0154] It should be understood that each component, compound, composition or parameter disclosed herein should be interpreted as disclosed for use alone or in combination with one or more of the various and each other components, compounds, compositions or parameters disclosed herein. It should also be understood that each amount / value of each component, compound or parameter disclosed herein or the range of its amount / value should be interpreted as also disclosed in combination with each amount / value or the range of its amount / value disclosed by any other component, compound or parameter disclosed herein, and therefore, for the purpose of this specification, any combination of the amount / value of two or more components, compounds or parameters disclosed herein or the range of its amount / value is also disclosed in combination with each other. Any and all features described herein and the combination of such features are included within the scope of the present invention, provided that these features are not mutually inconsistent.

[0155] Should be understood that, for identical component, compound or parameter, each lower limit of each scope disclosed herein should be interpreted as each upper limit combination of each scope disclosed herein.Therefore, the disclosure of two scopes should be interpreted as the disclosure of four scopes derived by combining each lower limit of each scope with each upper limit of each scope.The disclosure of three scopes should be interpreted as the disclosure of nine scopes derived by combining each lower limit of each scope with each upper limit of each scope etc.In addition, the specific amount / value of component, compound or parameter disclosed in specification sheet or embodiment should be interpreted as the disclosure of lower limit or upper limit of scope, and therefore can be combined with any other lower limit or upper limit or scope or specific amount / value of identical component, compound or parameter disclosed elsewhere in the application, to form the scope of this component, compound or parameter.

[0156] Example

[0157] Example 1. Comparison - Use of double stage distillation with the lipase process according to the invention

[0158] The starting oil in this example is mackerel oil, which has been deacidified (to remove free fatty acids) and stripped by a single column short-path distillation to remove organic contaminants. Refining is optional and can be done in different ways, such as pre-bleaching, using supercritical extraction, membrane filtration, etc. The same description applies to the other examples.

[0159] For comparison, batch 1 was prepared by ethylating fish oil (starting oil) by chemical ethylation of fish oil using 2% sodium ethoxide / ethanol in quantitative yield of ethyl ester (>95%). Conventional workup was performed by removing glycerol after evaporating excess ethanol and then washing the ethyl ester with water and citric acid. 2 The esters were distilled in a VTA short-path distillation pilot plant (VK 83-6) consisting of columns with an area of ​​1.3 m and a height of 0.285 m. The flow rate was adjusted to about 4 kg / hour, and the temperature on the first column was 115°C, while the temperature on the second column was 165°C. During the distillation, the pressure was less than 10 -3 The distillation was adjusted to remove mainly short chain fatty acids (C14 to C16) in order to maintain the content of long chain fatty acids (Table 1).

[0160] Batch 2 was prepared by selective transesterification according to the invention using 1,3 regiospecific lipase using stoichiometric amounts of ethanol. The lipase was 2% (w / w) of the oil. When the target conversion (change) was reached, the lipase was filtered off. Excess ethanol was removed and the ethyl esters were collected by single column stripping using the same VTA pilot plant as described above. The temperature on the stripping column was 165°C and the pressure was less than 10 -3 millibar.

[0161] Results from both batches are shown in Table 1 below. It should be noted that for both batches, the composition of cis-11-eicosenoic acid and cetyl oleic acid is very similar, 12% to 13% (121 to 123 mg / g) and 20% to 21% (199 to 203 mg / g), respectively. However, significant differences in the amount of EPA and DHA were observed between these batches. In batch 1, the total EPA+DHA was 26%, and EPA / DHA was 0.68, which is similar to the ratio present in the starting oil. In batch 2, using lipase, the total EPA+DHA was only 7%, and the ratio EPA / DHA was 4.4, which is more than six times the increase of the starting oil. Cetyl oleic acid / DHA is 16.04, which is more than 11 times the increase of the starting oil.

[0162] The results provided in the table further show that the amount of short chain fatty acids (C14 to C16) is about 27% (w / w) when lipase is used, compared to 7% when only distillation is used. This is a significant advantage of intermediates that can be further concentrated by distillation, because these short chain fatty acids have a relatively low boiling point, which will be reflected in the overall yield of the oil. This example shows that by using a 1,3-region specific lipase characterized by a certain fatty acid selectivity, a similar LCMUFA composition compared to distillation can be obtained, however, it contains much less EPA+DHA, and even less than the expected DHA compared to EPA. Those skilled in the art will recognize that the separation of fatty acids or their derivatives can be achieved because of their polarity (unsaturation) or boiling point. The skilled person will recognize that it is not trivial to separate LCMUFA such as cetyl oleic acid and cis-11-eicosenoic acid from EPA and DHA, and the results obtained in this example are unexpected and new.

[0163] Table 1 : Comparison of mild distillation of ethyl esters of mackerel oil by short path distillation after non-regiospecific chemical ethylation (2% sodium ethoxide / ethanol) and preparation of ethyl esters by selective transesterification with ethanol using a 1,3-regiospecific lipase (process of the invention).

[0164]

[0165]

[0166] Example 2. Concentration of cetyl esters by distillation with or without lipase

[0167] This example demonstrates how the ethyl esters prepared by selective ethylation according to the present invention unexpectedly concentrate cetyl esters more efficiently than non-selective ethylation. The two batches prepared in Example 1 were further distilled using the same pilot plant as in Example 1. Fractions were collected and analyzed by GC for estimation of mg / g and area % of key fatty acids. The results are shown in Table 2 below.

[0168] When batch 2 and batch 1 from Example 1 were concentrated, fractions A (selective / lipase) and C (non-selective / chemical) were prepared with comparable oil yields or 45% vs 51% oil yields, respectively. The sum of 20:1+22:1 was similar in both cases, 47% and 42%, respectively. It is important to note the significant differences between the two processes in EPA+DHA and cetylene / DHA ratios; 11% and 12.23 in fraction A compared to 31% and 1.37 in fraction C. Second, fraction A was still significantly richer in short-chain fatty acids (C14 to C16 and C18) compared to C, which favored further enrichment in cetylene acid. Therefore batch 2 (lipase) was further concentrated, resulting in fraction B with a total yield of 31%. This fraction had similar amounts of short-chain fatty acids as seen in C (9.8% vs 13.9%). However, fraction B was much richer in cetyl acid than that present in fraction C, with 44% cetyl acid EE (or 423 mg / g EE) compared to only 27% (265 mg / g EE) for the non-selective process. Still, only 12% EPA+DHA remained in fraction B and a high ratio of cetyl acid / DHA, or 13.08, which was 9.5 times higher than that present in fraction C and 9.6 times higher than that present in the starting oil.

[0169] It is further noted from Table 2 that the cetylene acid yields of fractions B and C are comparable, 83% and 87%, respectively, despite B containing about 1.6 times higher amounts of said fatty acid. This demonstrates that the lipase process of the invention is more efficient in producing high concentrations of cetylene acid while still maintaining low amounts of EPA+DHA and a high ratio of cetylene acid / DHA. The ratio of cis-11-eicosenoic acid / cetylene acid remains similar to that seen in the starting oil.

[0170] The skilled person will recognize that the comparative fraction C can be further distilled to a cetoleic acid content similar to that present in fraction B. However, this will result in significantly lower oil and cetoleic acid yields. Due to the similar boiling points of these fatty acid esters, further distillation of C will result in a proportional increase in the content of DHA and cetoleic acid. Therefore, such a cetoleic acid concentrate will be rich in cetoleic acid and DHA and have a low ratio of cetoleic acid / DHA. Therefore, this example shows that the use of a 1,3 regiospecific lipase characterized by a certain fatty acid selectivity is unexpectedly more effective in concentrating cetoleic acid compared to distillation.

[0171] Table 2: Comparison of concentrates of ethyl esters of cetyl esters prepared by short path distillation following non-regiospecific chemical ethylation (2% sodium ethoxide / ethanol) with the production of ethyl esters by selective transesterification with ethanol using a 1,3-regiospecific lipase.

[0172]

[0173]

[0174] Example 3. Concentration of cetyl esters by using herring oil and 1,3 regiospecific lipase.

[0175] Refined herring oil was treated with selective transesterification using a 1,3 regiospecific lipase and using stoichiometric amounts of ethanol and lipase. When the target conversion (conversion) was reached, the lipase was filtered off. Excess ethanol was removed and the ethyl esters (fraction A) were collected by single column stripping at 165°C. The distillate was further distilled by a two column short path distillation apparatus using the same VTA used in Example 1. During the distillation, the pressure was less than 10 -3 mbar and during the double distillation (concentration of cetyl esters) the column temperature was between 100° C. and 140° C. The results can be seen in Table 3 below.

[0176] After distillation of fraction A, an oil containing 44% cetoleic acid (434 mg / g EE) is harvested, which contains only 10% EPA+DHA, and the ratio of EPA / DHA is 4.72, which is 5.2 times higher than that present in refined menhaden oil. The ratio of cetoleic acid / DHA also significantly increased from 2.22 in the starting oil to 26.49 in the 44% cetoleic acid concentrate, which is about a 12-fold increase. The ratio of cis-11-eicosenoic acid to cetoleic acid is similar to that seen in the starting oil. Therefore, the method maintains the ratio while concentrating cetoleic acid. Therefore, the method of the present invention can be used for different types of cetoleic acid-rich fish oils.

[0177] Table 3: Selective transesterification of refined menhaden oil using a 1,3-regiospecific lipase and following distillation.

[0178]

[0179]

[0180] Example 4. Determination of the effects of LCMUFA concentrates on skin health including wound healing using human fibroblasts Research

[0181] Different fish oils contain different amounts of ω-3 fatty acids and other less well-known fatty acids. North Atlantic fish oil contains lower levels of EPA and DHA than South American fish oil, but is richer in long-chain monounsaturated fatty acids (LCMUFA), especially C22:n1-11 (cetylene acid) that is unique to marine oil. Compared with EPA and DHA, there are few studies on the intake, incorporation into tissues and health benefits of supplementing marine LCMUFA. Early studies (Huang et al, Cosmetic and Therapeutic Applications of Fish Oil's Fatty Acids on the Skin. Mar. Drugs 2018, 16, 256) showed that fish oil-derived fatty acids taken as oral supplements can improve skin barrier function, reduce UV-induced inflammation, reduce undesirable hyperpigmentation and improve wound healing. Several studies have shown the positive effects of fish oil and ω-3 on skin diseases, however, there are also several contradictory reports from meta-analysis. These contradictory results may be explained by the complex components and composition of fish oils, in addition the quality of the oils may vary depending on the fish species used and the manufacturing process. Both the LC-MUFA and omega-3 content of fish oils are important factors in determining the health effects of fish oils. Studies conducted by the applicant and NOFIMA in rats and salmon showed that for both species the highest concentration of the fatty acid C22:1n-11 (cetylene acid) was observed in the skin.

[0182] Preliminary cellular studies using human fibroblasts have been conducted to test the potential health benefits of LCMUFAs in the skin.

[0183] The primary objective of this study was to show whether adding LCMUFA concentrate (cetylene acid concentrate) or DHA to cell culture media would:

[0184] 1) affecting the ability of cells to regenerate after injury (the so-called "wound healing model"),

[0185] 2) Affects the production of melanin (skin pigment),

[0186] 3) Affects the expression of genes involved in inflammation following induction of oxidative stress.

[0187] The LCMUFA concentrate used in this study was a North Atlantic fish oil concentrate prepared from mackerel oil and contained:

[0188] Cetyl esters, C22:1: 48.74%,

[0189] DHA, C22:6:n3: 3.96%,

[0190] C20:1:18.84%,

[0191] Given as GC area %.

[0192] The concentration of EPA(C20:5n3) in the test material used in this study was 0.80 GC area %.

[0193] Wound healing model: Treatment arms included the addition of DHA or LCMUFA concentrate to the cell culture medium, respectively. Controls were cells supplemented with culture medium only without fatty acids. Effects on wound closure were measured 14 hours after wound initiation.

[0194] The results are shown in the accompanying drawings. Figure 1 , the Y axis shows the scratch wound area %, and the x axis provides bars for the control, DHA, and LCMUFA concentrates. The initial wound area was defined as 100%, and then the percentage of wound opening to the original wound was measured after 14 hours. Respectively, the reduction for LC-MUFA was about 60% and the reduction for DHA was about 50%. Therefore, a trend of improved wound healing was observed in the LCMUFA study group.

[0195] Menadione was added to the cell culture medium to induce oxidative stress. Melanin production was reduced in cells treated with both LCMUFA and DHA, see Figure 2 , which shows melanin concentration after oxidative stress.

[0196] The expression of genes related to inflammation did not show statistically significant differences between the treatment groups, but a trend towards lower expression of genes MMP2, ATF6B, and TNF-α was observed in the LCMUFA group compared to the control, see Figure 3 DHA-treated cells did not show such a trend and the data are not shown.

[0197] Conclusion: Preliminary data from cell studies suggest positive effects of LCMUFA concentrates on skin health, particularly undesirable hyperpigmentation due to decreased melanin synthesis.

[0198] Example 5. A randomized, double-blind nutritional study to determine the effect of concentrated cetyl ester fish oil on atopic dermatitis Research.

[0199] A study is being conducted to initially determine whether daily intake of 2g of the investigational product "EPAX Cetoleic 30" changes the grade and severity of those with mild to moderate atopic dermatitis as assessed by the Eczema Area and Severity Index (EASI). Secondarily, the objective is to determine whether daily intake of 2g of "EPAX Cetoleic 30" is associated with changes in molecular markers (lipid mediators, inflammatory markers, and blood lipids) in those with mild to moderate atopic dermatitis compared to placebo.

[0200] Research products:

[0201] North Atlantic fish oil concentrate, (EPAX Cetoleic 30) comprising at least 300 mg / g cetoleic acid. In particular, the product comprises the following fatty acids given in wt% and as GC area%:

[0202] EPA (C20:5): 5.7 wt%, 6.12 area%

[0203] DHA (C22:6): 1.6 wt%, 1.66 area%

[0204] Cis-11-eicosenoic acid (C20:1n9): 21.6% by weight, 23.16% by area

[0205] Cetearyl acid (C22:1n11): 41.5 wt%, 45.46 area%.

[0206] The total LCMUFA wt% was 67.6% (TG).

[0207] The investigational product is prepared by the claimed method.

[0208] The product is provided as 1 gram capsules. Subjects will take 2 x 1g capsules daily for 6 months. The placebo is corn oil.

[0209] The study population is based on volunteers (aged 18 to 80 years) with mild to moderate atopic dermatitis, with an Eczema Area and Severity Index (EASI) of 1.1 to 21.0. The project will use medical records from Norwegian hospitals for prescreening of subjects with mild to moderate atopic dermatitis. The study population will be randomized to ensure equal representation of medicated and unmedicated volunteers in each group.

[0210] The study is double-blind. According to the randomization scheme, subjects will be assigned to one of 2 nutrition groups, with 30 subjects in each group.

[0211] Group 1: Placebo: corn oil capsules.

[0212] Group 2: Study Supplement: EPAX Cetoleic 30-Capsules

[0213] Inclusion Criteria: Subjects diagnosed with active atopic dermatitis and EASI of 1.1 to 21.0, who were willing to avoid oil supplements (omega-3, borage, evening primrose, etc.) 1 month prior to study initiation (washout) and during the study, and were willing to take study supplements for 6 months.

[0214] Study visits will include clinical evaluations, blood sampling, skin sampling, and subject evaluations (questionnaires) at Day 1, after 6 weeks, after 3 months, and after 6 months.

[0215] Samples and analysis:

[0216] Clinical Assessment: Eczema Area and Severity Index (EASI)

[0217] Patient-reported assessments: Numerical Rating Scale (NRS), Dermatology Life Quality Index (DLQI), Patient Oriented Eczema Measure (POEM), use of surface treatments.

[0218] Skin strip samples (10 consecutive strips): ceramide, cytokines

[0219] Skin Barrier: Transepidermal Water Loss (TEWL)

[0220] Blood samples (Red blood cell (RBC) and plasma): ω-3 index, hydroxylated ω-3s

[0221] Blood samples for lipid profile (cholesterol profile) and liver enzymes (ALAT, ASAT, GTT) The primary efficacy variables will be changes in clinical and subject-assessed parameters for evaluation of eczema.

[0222] Secondary efficacy variables were:

[0223] Changes in ceramide / lipid composition in the skin

[0224] Changes in inflammatory signaling molecules in the skin

[0225] Changes in the ω-3 index in erythrocytes

[0226] Changes of cetyl ester in erythrocytes

[0227] Changes in cholesterol / lipid profile

[0228] Changes in transepidermal water loss (TEWL)

[0229] Changes in plasma hydroxylated ω-3 fatty acids

[0230] Correlation between ω-3 index and EASI parameters

[0231] Correlation between cetyl ester content in RBC and EASI parameters

[0232] When the study is finally completed, it is expected that at least one of the parameters used for eczema assessment will show a favorable change for subjects in Group 2 receiving capsules enriched with cetylene acid compared to Group 1 receiving placebo.

[0233] Example 6. Animal studies to assess effects on metabolic syndrome.

[0234] The study was conducted using ZDSD rats, an obese-diabetic rat model that spontaneously develops type 2 diabetes. The rats were fed a diet supplemented with herring oil (containing cetyl esters) for 6 weeks to assess how it affects the metabolic syndrome.

[0235] Supplementation of the diet with herring oil was found to improve insulin sensitivity and glucose levels compared to the soy control. These parameters are key targets for diabetes regulation and metabolic syndrome.

[0236] Future studies are also planned with Zucker fa / fa rats, which are naturally prediabetic, meaning they have mild insulin insensitivity and elevated fasting glucose levels, but lower than seen in rats with full-blown diabetes. The rats will be divided into a low-cetolyl group (herring oil), a high-cetolyl group (EPAX Cetoleic 30, the same investigational product used in Example 5), and a soy control group and dosed for 6 weeks. The study will measure parameters related to diabetes (glucose tolerance, insulin sensitivity, glucose uptake in muscle), diabetic neuropathy - especially allodynia (temperature sensitivity), and metabolic syndrome (liver fat content), as well as cardiovascular health (blood pressure).

Claims

1. LCMUFA composition comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), of which: The concentration of C22:1n11 is at least 16.0 wt %; EPA concentration of at least 3% by weight; The DHA concentration is a maximum of 4% by weight.

2. The LCMUFA composition of claim 1, which is derived from fish oil, in particular from oil of herring, mackerel, capelin, cod, shad or tobis.

3. The LCMUFA composition of claim 1 or 2 comprising 16.0 wt% to 60.0 wt% cetylene acid.

4. The LCMUFA composition of any one of claims 1 to 3, further comprising at least one C20:1 fatty acid, and wherein the weight ratio between the C22:1 and C20:1 fatty acids is from 1.5:1 to 2.6:

1.

5. The LCMUFA composition of any one of claims 1 to 4, comprising EPA at a concentration of 5.0% to 40.0% by weight of the composition.

6. The LCMUFA composition of any one of claims 1 to 5 comprising cis-11-eicosenoic acid C20:1n9 in the range of 9% to 28% by weight of the composition.

7. The LCMUFA composition of any one of claims 1 to 6, wherein the EPA:DHA ratio is at least 4:

1.

8. The LC-MUFA composition of any one of claims 1 to 7, wherein: The concentration of C22:1n11 is at least 39%, and The concentration of EPA is at least 5.0%.

9. The LCMUFA composition of any of the preceding claims, wherein the combined concentration of EPA and DHA ranges from 6.0% to 12.0% by weight of the composition.

10. The LCMUFA composition of any preceding claim, wherein the cetyl ester:DHA ratio is at least 10:

1.

11. A method for preparing a composition of LCMUFAs, the method comprising the steps of: i) mixing crude oil with lipase to obtain a mixture comprising LCPUFAs in the form of glycerides and LCMUFAs in the form of ethyl esters; ii) subjecting the mixture of step i) to distillation to obtain a LCMUFA composition as one fraction and a LCPUFA composition as another fraction.

12. The method of claim 11 for preparing a LCMUFA composition comprising C22:1n11 (cetylene acid) and C20:5n3 (EPA), wherein: The concentration of C22:1n11 is at least 16.0 wt %; EPA concentration of at least 3% by weight; The DHA concentration is a maximum of 4% by weight.

13. The method of claim 11 or 12, which produces a composition enriched in LCMUFAs wherein the concentration of cetyl esters is at least 39% and the concentration of EPA is at least 5.0%.

14. The method of any one of claims 11 to 13, wherein the crude oil is derived from herring, mackerel, capelin, cod, shad or tobis.

15. The method of any one of claims 11 to 14, wherein the fatty acids of the crude oil are primarily in the form of triglycerides.

16. The method of any one of claims 11 to 15, wherein the lipase is a 1,3-specific lipase.

17. The method of any one of 11 to 16, wherein the lipase is selected from the group consisting of Rhizomucor miehei lipase, Aspergillus niger lipase, Thermomyces lanuginosus lipase, Candida antarctica lipase, Candida rugosa lipase, Geotrichum candidum lipase, Penicillium roguefortii lipase, Rhizopus delemar lipase and Rhizopus oryzae lipase.

18. Use of the LCMUFA composition according to any one of claims 1 to 10 in food or feed.

19. The LCMUFA composition according to any one of claims 1 to 10 for use in therapeutic or health indications.

20. The LMCUMFA composition for use according to claim 19, for treating one or more of the following: metabolic syndrome, insulin sensitivity, fatty liver disease (NAFLD, NASH), diabetes, prediabetes, diabetic neuropathy, elevated LDL-cholesterol, cardiovascular diseases and risk factors, and skin diseases or conditions.

21. LCMUFA composition for use according to claim 19 or 20, wherein the use is in skin health or skin treatment.

22. A LCMUFA composition for use according to any one of claims 19 to 21, wherein the skin treatment aids the barrier function of the skin, maintains the skin's healthy appearance, avoids skin wrinkles or erythema, or protects the skin from the negative effects of sunlight UV radiation.

23. LCMUFA composition for use according to any one of claims 19 to 22, wherein the skin diseases and conditions are selected from the group consisting of eczema, psoriasis, dermatitis, acne and rosacea (papulopustular rosacea).

24. The LCMUFA composition for use according to claim 19 or 20, which is contained in the following: a pharmaceutical composition; a supplement, i.e. a food supplement, a nutritional supplement or a dietary supplement; a medical food; or a special medical purpose formula food.

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