Classifications

• • C—CHEMISTRY; METALLURGY
  • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
  • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
  • C11B3/00—Refining fats or fatty oils
  • C11B3/006—Refining fats or fatty oils by extraction
• • C—CHEMISTRY; METALLURGY
  • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
  • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
  • C11B3/00—Refining fats or fatty oils
  • C11B3/02—Refining fats or fatty oils by chemical reaction
• • C—CHEMISTRY; METALLURGY
  • C11—ANIMAL OR VEGETABLE OILS, FATS, FATTY SUBSTANCES OR WAXES; FATTY ACIDS THEREFROM; DETERGENTS; CANDLES
  • C11B—PRODUCING, e.g. BY PRESSING RAW MATERIALS OR BY EXTRACTION FROM WASTE MATERIALS, REFINING OR PRESERVING FATS, FATTY SUBSTANCES, e.g. LANOLIN, FATTY OILS OR WAXES; ESSENTIAL OILS; PERFUMES
  • C11B3/00—Refining fats or fatty oils
  • C11B3/12—Refining fats or fatty oils by distillation

Definitions

• the present invention relates to a new method of purification of fatty acid mono- and/or diglycerides, of any origin and in particular those characterized by an at least partial esterification with long chain polyunsaturated fatty acids (PUFAs), in particular C18 - C22, preferably C20 - C22, and comprising 2-6 double bonds, preferably 5-6 double bonds.
• PUFAs long chain polyunsaturated fatty acids
• the purification method according to the invention allows to remove from such glycerides products that are structurally unrelated to them.
• These foreign products especially in the case of natural glycerides, essentially consist, certainly not from the point of view of their weight but for their extreme toxicity, of so-called "Persistent" Organic Pollutants (POPs) now ubiquitously present in the environment (land, agricultural land, marine and river waters, etc.).
• POPs Persistent Organic Pollutants
• compositions of glycerides there may be mentioned, for example, the polychlorinated-dibenzo-dioxins and polychlorinated dibenzo-furans, polychlorinated biphenyls, polybrominated-diphenyl ethers, polycyclic aromatic hydrocarbons, and other pollutants that are also usually present and extremely toxic.
• the glycerides of fatty acids (esters with glycerol) object of the purification method of the invention can be mono- and/or di- glycerides and are so called in function of their degree of substitution. They are also reported in the literature as acyl-glycerols and distinguished according to their position and their steric structure into monoacyl-glycerols in position sn-1 or sn-2 and diacyl-glycerols in position sn -1,2 or sn -1, 3.
• glycerides depend essentially on the number, position and type of the acyl radical present as a glycerol ester.
• the fatty acids contained in the glycerides may be short-chain or more often medium- or long-chain; by "medium- or long-chain fatty acids", fatty acids with 16 or more carbon atoms are hereby meant. It is also known that said acids may be saturated (such as stearic acid C18:0), or monounsaturated (such as oleic acid C18:1 n-9), or polyunsaturated, often containing from 2 to 6 double bonds, and generally belonging to the class of omega-6 (or n-6) acids such as arachidonic acid (ARA, C20:4 n-6) or to that of omega-3 (or n-3) acids such as e.g.
• omega-6 (or n-6) acids such as arachidonic acid (ARA, C20:4 n-6) or to that of omega-3 (or n-3) acids such as e.g.
• EPA eicosapentaenoic acid
• DHA docosahexaenoic acid
• DPA docosapentaenoic acid
• omega-3 acids and their ethyl esters have acquired in recent decades an increasing importance for their biological properties and their use as nutraceuticals, food and dietary supplements, foods for special medical purposes, and as drugs. The same is happening at present for their glycerides, considered more "natural” and better absorbed in the human body, which have the added advantage of being their predominant source of production.
• omega-6 acids are especially abundant in plant oils and seeds, while the omega-3 acids and in particular EPA and DHA have predominantly marine origin and are derived essentially from fish oils, also from aquaculture, or from “krill oils” or even from algae and other oleaginous microorganisms, or from "single cell fermentation” from selected strains of algae or other microorganisms.
• the mono-glycerides are instead mainly industrial products, used for their excellent emulsifying properties, and represent 70-80% of the world emulsifiers used in food products, cosmetics and pharmaceuticals. They are obtained usually, although not with high yields and with low purity, by chemical glycerolysis of triglycerides (natural oils and fats) at high temperatures and by alkaline catalysis.
• the glycerides substituted with polyunsaturated acyl groups are preferably subjected to hydrolysis or alcoholysis by enzymatic route, by means of lipases - also immobilized - selective for the structure of the acyl and for its position on the glycerol: it is thus possible to operate in very mild reaction conditions and - by virtue of the higher resistance to enzymatic cleavage of polyunsaturated acyl groups, compared to saturated and monounsaturated acyls - it is also possible to isolate compositions of glycerols that are still partially acylated and particularly enriched in polyunsaturated acids.
• the purification method of the present invention preferably relates, in addition to the saturated and monounsaturated glycerides, to the purification of mono- and di-glycerides of any origin and obtained by any route, or their mixtures, including natural oils, partially substituted with radicals of long-chain fatty acids, comprising 18-22 carbon atoms, preferably 20-22 carbon atoms, and of polyunsaturated type, i.e. comprising 2-6 double bonds, preferably 5-6 double bonds .
• acids belonging to the class of omega-6 and/or omega-3 acids more preferably to the class of omega-3 acids, in particular to EPA and / or DHA.
• the content of a polyunsaturated fatty acid as defined above, e.g. EPA or DHA, or of their sum, is between 15-45%, more often 15-30%, and 90%, or between 30-60%, more often 45-60%, and 90%, of the weight of the composition , being 15-30% the average content in EPA or DHA or of their sum in a natural oil (fish oil or the like) used for the production of PUFAs, being 45% and 60% the minimum content of the sum of EPA and DHA and respectively of the sum of omega-3 acids - - in accordance with the monograph 01/2009: 1352 relative to the EP7.0 glycerides of omega-3 acids, reconstituted by chemical or enzymatic route and enriched in PUFAs, and being 90% around the maximum possible content for a glyceride having as substituents only polyunsaturated acids as described above, preferably EPA and/or DHA.
• glycerides derive, directly or indirectly, from "natural” and thus apparently reassuring sources, however poses serious health hazards in all uses mentioned above, due to the known presence of many environmental pollutants ("dioxins", polychlorinated biphenyls, etc..). Many of these are particularly fat-soluble and tend to concentrate together with the various fat components (from which the definition of Persistent Organic Pollutants, POPs).
• the main features of these substances are persistence over time, bio-accumulation through the food chain, the potential for dissemination and long-range environmental transport, and their toxicity. Many of these substances are clearly teratogenic, mutagenic and carcinogenic.
• the "dioxins” are in fact constituted by two chemical families of polychloro-dibenzo-para-dioxins (PCDDs) and of polychloro-dibenzo-furans (PCDFs): of these only 7 PCDDs and 10 PCDFs are of particular concern from a toxicological point of view.
• PCDDs polychloro-dibenzo-para-dioxins
• PCDFs polychloro-dibenzo-furans
• the PCDD/F are detected as mixtures of these toxic congeners, having attributed to each of them an appropriate "toxicity equivalent factor” (TEF), more precisely the WHO-TEF (European Commission Regulation (EC) 1881/2006, Off.J.EU, L364/5; 20 December 2006, p.20).
• TEF toxicity equivalent factor
• the maximum level of WHO PCDD/PCDF TEQ (Toxicity equivalent) for oils of marine origin intended for consumption in man according to the above Regulation (EC) 1881/2006, p. 18, is equal to 2 pg/g oil.
• acceptability criterion corresponds to "not more than” (NMT) 1 pg/g of WHO toxic equivalents.
• PCBs polychloro-biphenyls
• DL-PCBs dioxin-like PCBs
• TEF toxicity equivalent factors
• the PBDEs are polybromo- diphenyl ethers, constituted by molecules of diphenyl ether that are variously brominated, and some of them are highly neurotoxic and even carcinogenic. They are used as flame retardants, and are considered as persistent chemical pollutants of "emerging" type.
• the industrial production of some terms has already been banned, but their presence has been demonstrated in polluted waters and in landfill sites and in many samples of fish oils ( Zennegg M et al, Organohalogen Compounds, 68,1967,2006 ; US 7,732,488 ).
• IPAs or PAHs Polycyclic aromatic hydrocarbons
• PAHs are compounds formed by two or more fused aromatic rings and are of toxicological interest as they are considered possible carcinogenic. PAHs are usually expressed as marker substance benzo[a]pyrene, with a maximum permissible amount of 2 ng/g in oils and fats (Regulation (EC) 1881/2006, Section 6, p.18).
• patent application WO04/007654 ( EP1523541 and US 7,732,488 ) described a method for decreasing the amount of environmental pollutants in a mixture of oils or fats, by which a "working" volatile fluid is added to the mixture, then the mixture is subjected to at least one step of "stripping", during which a part of environmental pollutants together with the "working " volatile fluid is distilled off.
• the present invention relates to a method for the purification from persistent organic pollutants (POPs) of a composition consisting of mono-glycerides and/or di-glycerides of any origin, wherein the glycerol is at least partially esterified with long chain saturated or unsaturated fatty acids having 16-22 carbon atoms, this composition containing said POPs, wherein the method comprises the steps of:
• POPs persistent organic pollutants
• the mother waters and the washing waters of the above isolated urea complex can be pooled and treated, as a second purification step, with further 2-3 parts by weight of urea (with reference to the starting composition), repeating the steps from a) to c) of formation of the urea inclusion complex, cooling, isolating and obtaining the purified composition.
• the aforesaid long chain fatty acids have 18-22 carbon atoms, preferably 20-22 carbon atoms, and are of the polyunsaturated type (PUFA), i.e. containing 2-6 double bonds, preferably 5-6 double bonds, belonging to the class of omega-6 and/or omega-3 acids, preferably to the class of the omega-3 acids, more preferably represented by eicosapentaenoic acid (EPA, C20:5 n-3, all cis) and/or docosahexaenoic acid (DHA, C22:6 n-3, all cis).
• PUFA polyunsaturated type
• the composition comprising the above-mentioned mono-, and/or diglycerides is obtained by chemical and/or enzymatic route, by means of lipases selective for the fatty acid and its position on the glycerol, starting from natural triglycerides, or from glycerides or glycerol and polyunsaturated fatty acids, through known procedures.
• step a) of the method according to the present invention i.e. the treatment of the above-mentioned composition with urea, is performed by using as polar solvent a C 1 -C 4 alcohol or at least one C 1 -C 6 alkyl ketone or a mixture thereof.
• said polar solvent is selected from the group consisting of methanol, ethanol, n-butanol, i-butanol, acetone, methyl ethyl ketone and methyl isobutyl ketone.
• step a) of the method according to the present invention i.e. the treatment of the above-mentioned composition with urea, is performed by using as a polar solvent methanol in an amount equal to 4.5-7 parts by weight, or ethanol in an amount equal to 45-65 parts by weight.
• the environmental persistent organic pollutants (POPs) mentioned above comprise mainly polychloro-dibenzo-para-dioxins (PCDDs) and polychloro-dibenzo-furans (PCDFs), and/or polychloro-biphenyls (PCBs) dioxin-like (DL-PCBs) and markers, and/or polybromo- diphenyl ethers (PBDEs), and/or polycyclic aromatic hydrocarbons (PAHs), and their presence in the purified composition obtained in step (c) is at least 2 times reduced, preferably at least 5 times reduced, conveniently at least 10 times reduced, and in any case lower - for each component - than the respective limit of quantification (LOQ) with standard GC-MS method.
• PCDDs polychloro-dibenzo-para-dioxins
• PCDFs polychloro-dibenzo-furans
• PCBs polychloro-biphenyls
• DL-PCBs
• the present invention relates to a composition consisting of mono-glycerides and/or di-glycerides , as defined in claims 8, 9 and 17, as well as the use of such a composition for the preparation, from mono-glycerides and/or di-glycerides , of the corresponding polyunsaturated fatty acids or their salts, by chemical or enzymatic hydrolysis, or for the preparation of the related C1-C3 alkyl esters, as specified in claims 11 and 12.
• the present invention further relates to the use of a composition according to any of claims 8 and 9 for the preparation from said monoglycerides and/or diglycerides of the corresponding triglycerides, by means of chemical or enzymatic esterification or transesterification, in presence of said PUFAs as defined in claim 3 or of C 1 -C 3 alkyl esters thereof, preferably ethyl esters thereof.
• the present invention relates to a composition comprising these polyunsaturated fatty acids or their salts or C1-C3 alkyl esters obtained by chemical or enzymatic hydrolysis, as specified in claims - 13-15.
• the present invention relates to the use of a composition consisting of the above mentioned mono-glycerides and/or di-glycerides or the corresponding polyunsaturated fatty acids or their salts or C1-C3 alkyl esters for the preparation of formulations useful as food ingredients, food and dietary supplements, foods for special medical purposes (functional foods), food for animal consumption and for aquaculture, infant food formulas, cosmetic and pharmaceutical preparations, as specified in claim 16.
• a standard procedure provides that an oil or fat is hydrolyzed to give the acid components or transesterified in alcoholic medium to give the corresponding esters, by chemical or enzymatic route, and then one proceeds to the concentration of the polyunsaturated components: this generally occurs by molecular distillation, extraction with fluids under supercritical condition, or other methods, but more often by complexation precisely with urea ("inclusion" in urea).
• urea inclusion
• the use of urea is only presented as a means suitable for isolating and removing saturated and monounsaturated fatty acids, and therefore for the concentration of polyunsaturated acids in the solvent of the reaction: this is thus coupled - inter alia - to an increased concentration of extraneous impurities together with polyunsaturated components.
• acylglycerols appear to us absolutely poor, often conflicting, limited (since they only occasionally, if ever, comprise the PUFAs of highest interest, with longer chain and more unsaturated), and almost always bound to specific cases and therefore not generalizable.
• the ability of complexation is often attributed to a single or a few factors, while in our opinion it depends on all the reaction parameters in their entirety, as e.g. the degree of substitution of the glyceride, the length and the degree of unsaturation of the acyl, the ratio with the urea, the type and volume of solvent, the precipitation temperature of the solvent, and so forth, each factor interfering with all the others.
• a first aspect of the invention provides a method of purification which consists in treating the glyceride of a fatty acid - in a suitable solvent - with an excess amount of urea, sufficient for an essentially total complexation of the glyceride, generally greater than or equal to 3 parts by weight and up to 6 parts by weight or more, optionally subdividing the treatment in 2-3 consecutive steps, if necessary, preferably in 2 consecutive steps, and in any case until reaching an essentially total complexation.
• the division into multiple steps is not particularly harmful to the economy of the process since it involves only one additional filtration, in so far as it is possible to add the additional aliquot of urea directly to the filtered solution and the washing waters of the urea inclusion complex without further manipulation.
• the treatment is followed by a precipitation of the complex, optionally divided into the respective 1-3 fractions, then the complex is collected by filtration or centrifugation and washed thoroughly with the same solvent saturated with urea and precooled around 4-5°C or less, while the solvent containing the totality of the environmental pollutants (POPs) as described above and other POPs reported in the specialist literature, and any other impurities not covalently linked to the glyceride and not able to complex, are deleted. From the urea complex, or from the pooled fractions, the glyceride, which is now purified and essentially devoid of the abovementioned POPs, is then easily recovered.
• POPs environmental pollutants
• the glyceride may be a mono-and/or di-glyceride - (all also named as monoacyl- and diacyl-glycerols), and they will be at least partially esterified - in the case by far more preferred - with long-chain fatty acids, comprising 18-22 carbon atoms, preferably 20-22 carbon atoms, and of polyunsaturated type, i.e. comprising 2-6 double bonds, preferably 5-6 double bonds.
• glycerides may have any origin.
• omega-6 acids are particularly abundant in vegetable oils and seeds, while the omega-3 acids and in particular EPA and DHA have predominantly marine origin and derive essentially from fish oils, even from aquaculture, or from “krill oils” or still from algae and other oleaginous microorganisms, or from “single cell fermentation” starting from selected strains of algae or other microorganisms.
• the content of polyunsaturated fatty acids as defined above, especially EPA and/or DHA is higher than or equal to 15%, or is comprised between 15 and 90%, or in particular is comprised between 15 and 65%.
• the mono-glycerides and di-glycerides are instead mainly industrial products, and all glycerides enriched in polyunsaturated acids are obtained with chemical and/or enzymatic processes too.
• lipases that are selective for the fatty acid and its position on the glycerol are often used, starting from natural triglycerides to hydrolyze the saturated and monounsaturated acids, or from various glycerides or glycerol and reacting with concentrated polyunsaturated acids, and using known procedures.
• the content of polyunsaturated acids will usually be greater than or equal to 30%, or between 30 and 90%.
• this purification process shows some similarities with the process of enrichment of PUFA and their esters by complexation with urea, since it is possible to distinguish:
• step a) the use of urea in a strong excess and possibly in more steps
• step b) the careful washing of the complex to eliminate the pollutants contained in the mother liquors incorporated
• step c) the recovery of the purified product from the solid complex, instead of the recovery of the composition enriched in PUFA performed on mother waters as taught by the "prior art".
• the purified composition according to the invention will have essentially the same composition in glycerides and then in fatty acids as in the raw material, but will be substantially free from the totality of toxic environmental pollutants described above, such as dioxins and furans, polychlorinated biphenyls, polybrominated diphenyl ethers, cyclic and polycyclic hydrocarbons, simple and condensed, and other POPs known in the specialized literature.
• toxic environmental pollutants such as dioxins and furans, polychlorinated biphenyls, polybrominated diphenyl ethers, cyclic and polycyclic hydrocarbons, simple and condensed, and other POPs known in the specialized literature.
• each of the above-mentioned substances and their sum will be at least 2 times, or at least 5 times reduced, or preferably at least ten times reduced (reduction of at least 90%) with respect to the compositions from which they derive, if initially present in concentrations correspondingly higher than their limit of quantification (LOQ), or even each substance will be less than the limit of quantification (LOQ) with standard methodology GC-MS, whereby, by assimilating its value to "zero" (lower bound), we shall define their sum "essentially zero". A value of "zero” instead corresponds to a sum of data less than the limit of detection (LOD).
• LOD limit of detection
• the purified compositions of glycerides may be characterized by the contents of POPs that follow, although in most cases each congener will be generally lower than its limit of quantification LOQ (essentially zero).
• a second aspect of the invention relates to the compositions of glycerides themselves, as obtained according to the method of purification described above.
• compositions we explicitly mention a purified composition of mono- and/or di- esters of omega-3 acids with glycerol.
• a third aspect of the invention is that easily deducible from the availability itself of glycerides, free from environmental pollutants and highly purified: this aspect relates therefore to the use of said glycerides for the production by chemical or enzymatic route of other compositions based on PUFAs and omega-3 acids and derivatives, including PUFA-enriched triglycerides, in more or less concentrated form, according to standard procedures of the literature described in the field of the chemistry of these substances.
• the natural oils (triglycerides) more suitable for the intended purpose are subjected to a mild hydrolytic process, e.g. by alcoholic potassium hydroxide, thus obtaining the corresponding potassium salts and then the free acids and - if desired - the alkyl esters; or more often to a transesterification process e.g. in the presence of excess aliphatic alcohols, preferably C1-C3, and alkaline or acid catalyst, thus obtaining directly the corresponding alkyl esters of fatty acids, and from these - if desired - the related acids or salts.
• a mild hydrolytic process e.g. by alcoholic potassium hydroxide
• a transesterification process e.g. in the presence of excess aliphatic alcohols, preferably C1-C3, and alkaline or acid catalyst, thus obtaining directly the corresponding alkyl esters of fatty acids, and from these - if desired - the related acids or salts.
• the salts of the polyunsaturated acids are obtainable from these e.g. by direct salification and are represented by the salts with alkali metals, e.g. sodium and potassium, alkaline-earth metals, e.g. calcium, with basic amino acids such as lysine and arginine, meglumine, with choline and mono-, di-, and triethanolamine, and the like, if pharmacologically acceptable.
• alkali metals e.g. sodium and potassium
• alkaline-earth metals e.g. calcium
• basic amino acids such as lysine and arginine, meglumine, with choline and mono-, di-, and triethanolamine, and the like, if pharmacologically acceptable.
• alkyl esters are obtainable e.g. by chemical or enzymatic alcoholysis and are represented by esters with aliphatic alcohols, even very long chain alcohols as found in "natural waxes", but are preferably represented by esters with lower alcohols C1-C3, in particular the ethyl ester.
• the glycerides reconstituted and concentrated in PUFAs can be prepared by esterification e.g. of concentrated and purified omega-3 acids with glycerol, or by transesterification of esters with glycerol.
• a fourth aspect of the invention comprises the addition to the purified glycerides and derivatives thereof of suitable diluents, excipients, suspending agents, etc. and/or preservatives, antioxidants, etc., according to all the technologies known in the art, to give all the formulations known in the art to enable their use in all the proposed uses.
• Said formulations comprise, in addition to their direct inclusion e.g.
• compositions and their formulations will be addressed to the preparation and use as a food or food ingredient, of every kind and purpose, as food and diet supplement, food for special medical purposes (functional food), both new and deductible from the pharmaceutical use, feed for animal consumption and for aquaculture, infant food formulas, cosmetic and pharmaceutical preparation, all containing or enriched with long chain fatty acids or derivatives thereof, having polyunsaturated character or specifically of the omega-3 series.
• the preparation for pharmaceutical use will preferably be enriched and concentrated in polyunsaturated components, in particular in EPA and/or DHA, or salt thereof, or their ethyl ester, or their glyceride.
• compositions will be directed to the prevention and treatment of risk factors for heart diseases, cardiovascular and cardio-circulatory illnesses, such as hypertension, severe and moderate hypertriglyceridemia (resp. >500mg/dl and >200mg/dl) and hypercholesterolemia, in particular the familial and genetic forms, also in combination with other drugs and in particular with statins, and as the defects of coagulation and platelet aggregation.
• risk factors for heart diseases, cardiovascular and cardio-circulatory illnesses such as hypertension, severe and moderate hypertriglyceridemia (resp. >500mg/dl and >200mg/dl) and hypercholesterolemia, in particular the familial and genetic forms, also in combination with other drugs and in particular with statins, and as the defects of coagulation and platelet aggregation.
• a use of great importance is the one for the prevention and treatment of heart diseases, and cardiovascular and cardio-circulatory illnesses, such as coronary- atherosclerotic illnesses and cardiac and cerebral ischemic states, including myocardial and cerebral infarction, and for reducing the risk of sudden cardiac death consequent to myocardial infarction; those of electrical origin and involving the onset and the propagation of the heart rhythm, including arrhythmia and atrial and/or ventricular fibrillation; and those due to mechanical defects of the heart pump, such as heart insufficiency and decompensation and/or the "congestive heart failure".
• cardiovascular and cardio-circulatory illnesses such as coronary- atherosclerotic illnesses and cardiac and cerebral ischemic states, including myocardial and cerebral infarction, and for reducing the risk of sudden cardiac death consequent to myocardial infarction
• those of electrical origin and involving the onset and the propagation of the heart rhythm including arrhythmia and atrial and/or ventricular fibrillation
• compositions are used for the prevention and treatment of disorders of the central nervous system (CNS), including epilepsy, the various depressive forms, the bipolar disorders, the pediatric pathologies by defects of the attention and hyperactivity disorders (ADHD), defects of learning and memory, the various forms of schizophrenia, Alzheimer's disease and various forms of dementia.
• CNS central nervous system
• ADHD attention and hyperactivity disorders
• the memory enhancements particularly in pediatric age, are of extreme interest.
• Still other pharmaceutical uses include the prevention and the treatment of retinopathy and symptoms of dry eye, of the metabolic syndrome, defects of metabolism and related to obesity, type 2 diabetes, liver disorders, diseases of connective tissue and joints, of inflammatory states, autoimmune diseases, ulcerative colitis, psoriasis and tumor disease.
• the final aspect of the present invention finally concerns the purification of a composition consisting of a mono-glyceride and/or diglyceride of any origin, esterified with long-chain fatty acids consisting of 16-20, preferably 18-20 carbon atoms, of type saturated and/or monounsaturated, and the purified composition which is obtained.
• the purification from POPs was carried out on a composition containing a mixture of monoglycerides (52.4%) and diglycerides (46.0%) of polyunsaturated fatty acids (PUFAs).
• PUFAs polyunsaturated fatty acids
• composition has been obtained by direct esterification with glycerol of a concentrated composition at more than 80% of n-3 PUFAs derived at their turn from their corresponding ethyl esters (batch 201308, origin from fish oil; GC/MS analysis: EPA 51% b.w., DHA 39% b.w.) by standard alkaline hydrolysis (KOH in aqueous ethanol and reflux for 2 hours).
• glycerides The synthesis of glycerides was carried out in the presence of a preparation of immobilized lipase from Candida antarctica (NV-435) and of molecular sieves, and the ratios of components, the temperatures and the reaction times adopted were those reported for the experiment 2 of Noriega-Rodriguez J.A. et al, J. Food Res. 2(6),97, 2013 .
• the mixture is left to cool with stirring and at about 45°C quickly starts the precipitation of a precipitate, which is then cooled for a few minutes with running water and then left to rest for 1 hour in a refrigerator at about 5°C to complete the precipitation.
• the collected precipitate is washed thoroughly on the buchner with two 50 mL portions of a urea solution in methanol, obtained by dissolving 16 g of urea in 100 mL of methanol under heating and then cooling to 5°C, thus obtaining at the end colorless washing waters.
• the two organic phases are separated from the aqueous phase and pooled, then the n-hexane is evaporated to dryness at about 35°C and under reduced pressure up to constant weight, obtaining 20.8 g of residue consisting of refined mixture of mono- and diglycerides that is kept around 5°C in a nitrogen atmosphere.
• methanol mother liquors isolated by filtration of the urea inclusion complex of step 1, containing eventual butanol and combined with the methanol washing as described above, are added with a further 50 g of urea (total ratio by weight glycerides : urea of 1 : 6) and brought to the boiling for 15 minutes under stirring, obtaining almost complete solution.
• the precipitate is filtered on a buchner, the mother liquors are thoroughly removed and the precipitate is washed carefully with 2 portions of 50 mL of a urea solution in methanol obtained as described above and pre-cooled to 5°C.
• the purification from POPs was performed on a composition containing substantially only monoglycerides (98.5%) of PUFAs, obtained as above by direct esterification with glycerol of the same above concentrated composition of n-3 PUFAs.
• the content of PUFAs in the monoglyceride was EPA 53.5% b.w. and DHA 38%.

Landscapes

• Chemical & Material Sciences (AREA)
• Life Sciences & Earth Sciences (AREA)
• Chemical Kinetics & Catalysis (AREA)
• Microbiology (AREA)
• Engineering & Computer Science (AREA)
• Oil, Petroleum & Natural Gas (AREA)
• Wood Science & Technology (AREA)
• Organic Chemistry (AREA)
• General Chemical & Material Sciences (AREA)
• Coloring Foods And Improving Nutritive Qualities (AREA)
• Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)

Applications Claiming Priority (1)

Publications (1)

Family

ID=59521560

Family Applications (1)

Country Status (2)

Citations (5)

• 2017
  • 2017-03-24 IT IT102017000032907A patent/IT201700032907A1/it unknown
• 2018
  • 2018-03-19 EP EP18162570.8A patent/EP3378926A1/fr not_active Withdrawn

Patent Citations (7)

Non-Patent Citations (14)

Also Published As

Similar Documents

Legal Events