WO2007123425A1 - PROCÉDÉ PERMETTANT DE SÉPARER des MATIÈRES lipidiques - Google Patents

PROCÉDÉ PERMETTANT DE SÉPARER des MATIÈRES lipidiques Download PDF

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Publication number
WO2007123425A1
WO2007123425A1 PCT/NZ2007/000088 NZ2007000088W WO2007123425A1 WO 2007123425 A1 WO2007123425 A1 WO 2007123425A1 NZ 2007000088 W NZ2007000088 W NZ 2007000088W WO 2007123425 A1 WO2007123425 A1 WO 2007123425A1
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Prior art keywords
feed material
product
solvent
comprises greater
ethanol
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David Stevenson
Dawn Scott
Owen John Catchpole
Andrew Mackenzie
Mikhail Vyssotski
Vasily Svetashev
Tina Fenton
Jason Murney
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    • 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
    • C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
    • C11B7/0008—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents
    • C11B7/005—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents in solvents used at superatmospheric pressures
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/23—Removal of unwanted matter, e.g. deodorisation or detoxification by extraction with solvents
    • 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
    • C11B7/00—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils
    • C11B7/0008—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents
    • C11B7/0025—Separation of mixtures of fats or fatty oils into their constituents, e.g. saturated oils from unsaturated oils by differences of solubilities, e.g. by extraction, by separation from a solution by means of anti-solvents in solvents containing oxygen in their molecule
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • This invention relates to fractionation processes. More particularly it relates to a process for fractionating a feed material containing phospholipids (including for example phosphatidyl serine, cardiolipin or sphingomyelin), gangliosides, or a combination thereof.
  • phospholipids including for example phosphatidyl serine, cardiolipin or sphingomyelin
  • gangliosides or a combination thereof.
  • Phospholipids are major components of all biological membranes, and include phosphoglycerides (for example phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI), cardiolipin (CL) and phosphatidyl serine (PS)); and sphingolipids such as sphingomyelin (SM).
  • PC phosphatidyl choline
  • PE phosphatidyl ethanolamine
  • PI phosphatidyl inositol
  • CL cardiolipin
  • PS phosphatidyl serine
  • SM sphingomyelin
  • Gangliosides are components in the cell plasma membrane which modulate cell signal transduction events. They are implicated as being important in immunology, brain function and neurodegenerative disorders.
  • Both phospholipids and sphingolipids are involved in cell signalling events leading to, for example, cell growth, cell differentiation, cell proliferation, and programmed cell death (apoptosis).
  • MFGM bovine milk fat globule membrane
  • Both phospholipids and sphingolipids have been implicated in having a number of health benefits, including brain health, sports nutrition, skin health, eczema treatment, anti- infection, wound healing, gut microbiota modifications, anti-cancer activity, alleviation of arthritis, improvement of cardiovascular health, and treatment of metabolic syndromes.
  • the PI fraction was treated with acetone to remove neutral lipids.
  • Aqueous ethanol was used in concentrations of 84%, 89%, 95% and 99% at 5:1 solvent to lecithin ratio.
  • the optimal moisture content of lecithin-ethanol mixtures was found to be 24% which was achieved by using a 95% ethanol-to-lecithin ratio of 5 : 1.
  • Yield, purity and recovery of PC fraction increased with increasing temperature, with maximum purity achieved at 40 °C.
  • the optimal solvent-to-sample ratio was 6:1 (PC yield- and recovery wise) or 10:1 (for PI). Maximum purity of PC was achieved when fractionated twice, but the purity of PI fractions decreased with increasing number of fractionations.
  • the traditional approach used to obtain high PS fractions is not by enrichment of PS- containing mixtures, but rather by the use of phospholipase D, serine and PC or a phospholipid mixture containing PC (US 5,700,668 A, KR2003086128 A, and JP 2079990 A).
  • the phospholipase D enzyme is used to convert PC to PS.
  • JP 2002241385 A a mixture of phospholipids is dissolved in alcohol, followed by the addition of metallic salts to precipitate PS, while JP 3047192 A and JP 2805522 B2 disclose the use of centrifugal partition chromatography with an aqueous mixed solvent containing a saturated hydrocarbon, alcohol and ether to separate PE, PC and SM.
  • JP 8322472 A discloses a process for concentrating acidic lipids from plant (soybean) lecithin.
  • Plant-derived lecithin suitably soybean lecithin containing at least 40% phospholipids
  • a lower alcohol containing less than or equal to 15% water, and preferably less than or equal to 10% water, at over 30 0 C, and preferably at 35-80 0 C.
  • the lower alcohol fraction is then removed.
  • the phospholipids to lower alcohol ratio is preferably greater than 15 on a weight basis.
  • the treatment is preferably repeated 3-5 times where each treatment uses a phospholipids-to-low alcohol ratio of greater than one on a weight basis.
  • the resulting acidic phospholipid concentrate preferably contains more than 60% PI and phosphatide acid (PA).
  • JP 95151260 A and US 5,833,858 A describe a more elaborate approach wherein the first step comprises treating plant-derived lecithin with a solvent containing 85% to 100% of a lower alcohol and then eliminating the solvent fraction containing said lower alcohol to give a phospholipid mixture, and the second step comprises treating the said phospholipid mixture with an aqueous solution containing 75% or less of ethanol.
  • the water-soluble components contained in the phospholipid mixture obtained in the first step are efficiently removed in the second step.
  • a lipid mixture containing a high concentration of the acidic phospholipids PI and PA can be relatively easily obtained from plant-derived lecithin.
  • a process for concentrating PS, SM, CL or GS was not disclosed.
  • JP 10265485 A describes the use of ammonia-containing ethanol to prepare highly concentrated PA and PI mixtures, as well as PE and PC mixtures, from lecithin.
  • a further, more elaborate process is disclosed in US 5,214,171 A for fractionating phosphatide mixtures into two or more fractions which are enriched in one or more of PC, PE, PI and PA by carrying out extraction steps using an alcoholic solvent in which the solubilities of PC, PE and PA are controlled by suitably adjusting the acidity of the solvent, the pH being adjusted to above 8 for solubilizing PC and PE and to below 5 for solubilizing PA.
  • PI is substantially insoluble in the solvent used in the process and thus is mainly recovered in the extraction residue.
  • butanol- water- diisopropylether partitioning is an effective way to prepare GS-enriched fractions (Ladish & Gillard 1985). This method is applicable to the total lipid extracts of plasma, cells, or animal tissues. Partitioning of the dried total lipid extract is performed in a three-component solvent system consisting of diisopropyl ether, 1- butanol, and 50 M aqueous NaCl (6/4/5, v/v/v). Gangliosides partition nearly quantitatively into the lower aqueous phase, and other lipids into the upper organic phase resulting from the mixture of these three solvents. The ganglioside-containing aqueous phase is then freed of salts and other low-molecular-weight impurities by gel filtration.
  • gangliosides often involves heating ganglioside containing raw materials, like buttermilk or whey protein concentrate, under alkaline conditions (e.g. US 5,831,079, US 5,795,980, JP 93035155 B 5 JP 3615798 B2). It should be noted that under these conditions ganglioside GD3 is hydrolysed and forms ganglioside GM3.
  • Other commonly used approaches include ultrafiltration (e.g. JP 3176698 B2, WO 91/07417), column chromatography (e.g. DE4221190 Al, CA 2002155 A, IT 1252310 B) or both (e.g. JP 2207090 A, CN 1379034 A).
  • a composition containing at least 4% gangliosides was produced by adding ethanol to milk or milk products to achieve a final ethanol concentration of 30-70 vol% and recovering the precipitate; then by adding ethanol to the precipitate to give an ethanol concentration of 70- 90 vol%, removing the protein precipitate, and membrane filtering the ethanol fraction (US 5,844,104).
  • ganglioside-enriched compositions Preparation of ganglioside-enriched compositions is also described in US 6,265,555.
  • This patent claims a method of manufacturing a composition containing ganglioside from milk or a milk-derived material (butterserum, buttermilk, whey, and whey protein concentrate), comprising the steps of: dispersing a material containing ganglioside in an ethanol solution to make the concentration of ethanol 60-95%; heating said dispersion to a temperature of 50- 9O 0 C to dissolve ganglioside and simultaneously generate a first precipitate containing proteins; removing the first precipitate; cooling a supernatant excluding said first precipitate to a temperature lower than O 0 C to generate a second precipitate containing ganglioside; and recovering the second precipitate.
  • the present invention provides a process for fractionating a feed material into soluble and insoluble components, comprising
  • the feed material comprises less than 10% protein, and less than 10% lactose. More preferably less than 10% of the feed material comprises protein and lactose.
  • the feed material comprises greater than 1% phosphatidyl serine.
  • the feed material comprises greater than 2% phosphatidyl serine. More preferably the feed material greater than 5% phosphatidyl serine.
  • the feed material comprises greater than 1% sphingomyelin.
  • the feed material comprises greater than 5% sphingomyelin. More preferably the feed material comprises greater than 15% sphingomyelin.
  • the feed material comprises greater than 1% cardiolipin.
  • the feed material comprises greater than 2% cardiolipin. Most preferably the feed material comprises greater than 5% cardiolipin.
  • the feed material comprises greater than 0.3% gangliosides.
  • the feed material comprises greater than 1% gangliosides.
  • the feed material comprises greater than 2% gangliosides.
  • the feed material of the present invention may be derived from terrestrial animals, marine animals, terrestrial plants, marine plants, or micro-organisms such as microalgae, yeast and bacteria.
  • the feed material is derived from sheep, goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human.
  • the feed material is selected from: tissue, a tissue fraction, organ, an organ fraction, milk, a milk fraction, colostrum, a colostrum fraction, blood and a blood fraction.
  • the feed material is derived from dairy material, soy material, eggs, animal tissue, animal organ or animal blood. More preferably the feed material is selected from: a composition comprising dairy lipids, a composition comprising egg lipids, and a composition comprising marine lipids.
  • the feed material used in the process of the present invention is a bovine milk fraction.
  • the feed material is selected from: buttermilk, a buttermilk fraction, beta serum, a beta serum fraction, butter serum, a butter serum fraction, whey, a whey fraction, colostrum, and a colostrum fraction.
  • the feed material may comprise milk fat globule membrane.
  • the feed material may have been genetically modified.
  • the feed material is in solid form.
  • the feed material may be cryomilled before contact with the solvent.
  • the feed material used in the process of the present invention is cryomilled to achieve a mean particle size of 0.1 -5mm before contact with the solvent.
  • the solvent of the present invention preferably comprises:
  • the solvent comprises between 0 and 20% v/v water. Most preferably the solvent comprises between 1 and 10% v/v water.
  • the alcohol is ethanol.
  • the solvent used in the process of the present invention comprises 95% aqueous ethanol.
  • the ratio of solvent to feed material in the process of the present invention is between 10: 1 to 100: 1. More preferably the ratio of solvent to feed material is between 20: 1 to 60:1. Most preferably the ratio of solvent to feed material is between 25:1 to 40:1.
  • the process of the present invention is carried out at greater than 3O 0 C. More preferably the process of the present invention is carried out at greater than 5O 0 C. Most preferably the process of the present invention is carried out at greater than 55 0 C.
  • the process of the present invention may be carried out in a number of steps of decreasing or increasing temperatures.
  • the process of the present invention is carried out at less than or equal to 4 0 C. More preferably the process is carried out at less than or equal to O 0 C.
  • step (c) of the process of the present invention is carried out for between 1-3 hours. Most preferably step (c) is carried out for 2 hours.
  • the solvent and feed material are contacted once, the solvent and feed material may be contacted multiple times with the same batch of solvent, or the feed material may be contacted with more than one batch of solvent.
  • the solvent and feed material are contacted by using a packed bed of feed material with the solvent flowing through it.
  • the solvent and feed material are mixed by agitation.
  • the agitation is achieved by high shear mixing.
  • the invention also provides products produced by the process of the invention, both the insoluble components remaining after contact with the solvent (also referred to herein as the "residue”); and the soluble components that are dissolved in the solvent after contact with the feed material (also referred to herein as the "extract”). Where the feed material is contacted with more than one batch of solvent, or the solvent is cooled in a number of steps, there will be multiple "extract" products.
  • the product contains more sphingomyelin than the feed material.
  • the product comprises greater than 3% sphingomyelin. More preferably the product comprises greater than 10% sphingomyelin. Most preferably the product comprises greater than 15% sphingomyelin.
  • the product contains more phosphatidyl serine than the feed material
  • the product comprises greater than 5% phosphatidyl serine. More preferably the product comprises greater than 30% phosphatidyl serine. Most preferably the product comprises greater than 70% phosphatidyl serine.
  • the product contains more gangliosides than the feed material
  • the product comprises greater than 2% gangliosides. More preferably the product comprises greater than 4% gangliosides. Most preferably the product comprises greater than 6% gangliosides.
  • the product contains more cardiolipin than the feed material
  • the product comprises greater than 5% cardiolipin. More preferably the product comprises greater than 10% cardiolipin. Most preferably the product comprises greater than 25% cardiolipin.
  • PL means phospholipids
  • C cardiolipin (diphosphatidyl glycerol)
  • PC phosphatidyl choline
  • PI means phosphatidyl inositol
  • PA means phosphatidic acid
  • PS means phosphatidyl serine
  • PE means phosphatidyl ethanolamine
  • SM means sphingomyelin
  • DHSM dihydrosphingomyelin
  • GS means gangliosides
  • MFGM milk fat globule membrane
  • the present invention is based on recognising that there is a difference in solubility of phospholipids in short-chain monohydric alcohol solvents. Using this solubility differential, the inventors have discovered that the insoluble phospholipids (including PS and CL) and GS may be fractionated from the soluble phospholipids (including SM). A number of variables in this process can be manipulated to optimise the fractionation, as follows;
  • feed materials may be used in the present invention.
  • the feed material may be derived from sheep, goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama, bovine or human tissue, milk, colostrum, or blood.
  • the feed material may be derived from soy or other plant material, eggs, animal tissue or organs, marine animals, marine plants or micro-organisms.
  • the feed material may have been genetically modified.
  • a small particle size is preferred so that the total surface area is increased to allow the soluble phospholipids to dissolve.
  • Methanol, ethanol, n-propanol and iso-propanol and mixtures thereof are all suitable solvents, as are these alcohols with up to 40% v/v water.
  • 95% v/v aqueous ethanol is a preferred solvent as it results in the lowest solubility of PS, CL and GS and the highest solubility of SM.
  • the inventors have found that using a single solvent system to fractionate the phospholipids and/or gangliosides of interest simplifies fractionation processing.
  • the degree of agitation must be sufficient to keep the particles suspended and aid the extraction by breaking up any aggregates that form. Those skilled in the art will recognise that high shear mixing would be the most advantageous at a large scale.
  • the lipid-rich feed material is a solid
  • the solvent can be passed through the packed bed of ground feed material at a low temperature to extract compounds with the highest solubility (and lowest melting point), and then increasing the temperature (either stagewise or continuously) until the desired final temperature is achieved that gives the highest purity of PS, CL and/or GS in the residue.
  • This process uses temperature rather than the solvent to feed stream ratio to achieve fractionation.
  • PC is selectively dissolved.
  • Subsequent temperature steps can then be completed to remove the more soluble phospholipids (PE, SM and PI) up to a last step of between 50 and 60°C.
  • the resulting packed bed is enriched in PS and/or CL and gangliosides and is a powdery precipitate.
  • Example 1 One Pot Stirred Tank Ethanol Extraction of Dairy Feed Streams
  • Feed Stream A High in lipid content (up to 94% lipid by Modified Roese-Gottling), with approximately 32% phospholipids and 2.3% ganglioside. This Feed Stream was produced by near critical dimethyl ether extraction of a MFGM rich fraction of bovine buttermilk with reduced lactose content.
  • Feed Stream B Contains high levels of phospholipids, 65% by weight. This feed stream was produced by supercritical CO 2 extraction of the MFGM rich fraction of bovine buttermilk with reduced lactose content (to remove neutral lipids) prior to near critical dimethyl ether extraction.
  • Feed Stream C has a very high lipid content and a proportionally higher amount of phospholipids than Feed Stream A, but almost no gangliosides. This Feed Stream was produced by near-critical dimethyl ether extraction of a MFGM rich fraction of bovine buttermilk with reduced lactose content.
  • the phospholipids and ganglioside compositions of the three feed streams are outlined in Table 1 below.
  • Each feed stream was introduced to an excess of solvent (95% ethanol) at a temperature above 5O 0 C and agitated for a determined length of time. During this process the ethanol soluble phospholipids were dissolved into the ethanol producing an extract stream, and leaving behind the insoluble material ("residue") which as a result has enriched levels of PS and GS.
  • solvent 95% ethanol
  • Feed Streams A and C are the easiest to process. They are liquid at 4O 0 C and so can be pumped. They are also both solid at O 0 C so can be fed through a Shiver. This provides two options for introducing these feed streams to the solvent, because in either form (liquid or frozen), with ethanol at elevated temperatures, both dissolve readily.
  • Feed Stream B is a solid gum at all practical processing temperatures so it requires cryogenic milling before ethanol extraction. Thus for this process Feed Stream B needed to be milled to a fine powder (this required cryogenic milling) before introduction into the solvent. This additional processing step adds costs, making it significantly more expensive to process than Feed Streams A or C. There is also a risk of not milling Feed Stream B sufficiently, which would compromise the process and cause a lower achieved purity. However, this problem may be overcome by dispersing the feed stream in a small amount of solvent first. Feed Streams A and C are liquid at elevated temperatures because of the high neutral lipid content while Feed Stream B is solid at all practical temperatures because of its low neutral lipid content.
  • the syringe was then heated to 60 0 C to liquefy the feed stream.
  • the liquefied feed stream A was then added to the ethanol over a 5 minute period by depressing the syringe.
  • the solution was then passed through a Whatman 4 filter paper and 0.235 g of insoluble material (the residue) was collected.
  • the ethanol mixture that passed through the filter contained the remaining 2.265 grams of phospholipid material (the extract).
  • the ethanol was agitated by a magnetic stirrer (Jarike&Kunkel IKA Labortechnik RCT basic magnetic stirrer) until its temperature reached 60 0 C.
  • the syringe was then heated to 60 0 C to liquefy the feed stream. 5.
  • the liquefied feed stream A was then added to the ethanol over a 5 minute period by depressing the syringe.
  • This example shows a method for processing solid lipid-rich substrates.
  • the milled material was placed in a 1 L Schott bottle and 200 mL of 95% ethanol at a temperature of ⁇ 4 0 C was poured into the bottle
  • This example shows that PS is substantially enriched when the feed stream is low in GS and lactose.
  • This example shows that low processing temperatures result in low levels of enrichment of PS.
  • Feed Stream B was cryogenically milled, and placed in a 50 mL centrifuge tube.
  • Feed Stream B of Example 1 was also used in this example. As for Example 1, Feed stream B was cryogenically milled to a fine powder before introduction into the solvent.
  • the vessel was shaken periodically and the temperature was maintained at 65 0 C by placing the vessel in a hot water bath.
  • the remilled material was placed in a 5L vessel, and 5L of 95% ethanol at 65 0 C was added. The vessel was then shaken periodically and the temperature was maintained at 65 0 C by placing the vessel in a hot water bath.
  • the powder residue was placed in a 5L vessel and 5L of 95% ethanol at 65 0 C was added.
  • the vessel was shaken periodically and the temperature was maintained at 65 0 C by placing the vessel in a hot water bath.
  • Table 3 shows the results achieved by this process.
  • Table 3 Residue and extract fraction compositions from multistep extraction
  • Example 3 Packed Bed Ethanol Extraction of Solid Feed Streams
  • Dairy feed stream B was used, but the general method is applicable to soy lecithin, and phospholipids concentrates from animal organs and tissues.
  • the feed stream is cryogenically milled to a fine powder.
  • This process uses temperature rather than the solvent to feed stream ratio to achieve fractionation.
  • the resulting packed bed is enriched in PS and gangliosides and is a powdery precipitate
  • This process is also advantageous as at temperatures above 60°C PS becomes soluble and significant losses can be experienced.
  • Another advantage of this technique is the elimination of the need for high shear agitation. While agitation generally breaks aggregates up, high shear agitation may also bring the particles together in collisions that could form aggregates. This is a concern when less than optimal agitation is employed. For example it is a real risk in scale up where factors like tank diameter and height, mixing speed, impeller type and size come into play.
  • this process uses non mixing (laminar) gentle fluid dynamics to contact the feed and the ethanol, so there is minimal collision and hence less chance for aggregates to form.
  • a known amount of milled Feed Stream B (5.1062 grams) was spread evenly inside a jacketed sinter filter.
  • the sinter jacket was connected to a temperature controlled water bath with a pump.
  • the water bath was set to 10 0 C.
  • the water bath temperature was raised to 3O 0 C.
  • the water bath temperature was raised to 50 0 C.
  • the water bath temperature was raised 6O 0 C.
  • the extracts from each step can be combined or left as separate phospholipid fractions.
  • the 2O 0 C and 30 0 C extracts are at least 25% SM product, while others could be used as a PC/PE phospholipid mixture. Discussion of Examples 1-3
  • Feed Stream feed material
  • Example 1 Three different feed streams were used in Example 1.
  • Feed streams A and C proved easiest to use, as they are liquid at temperatures above 40°C and hence are dispersed into the solvent with ease.
  • Feed Stream B on the other hand is a gummy solid at temperatures above 0 0 C. Below 0°C the solid becomes rigid and cryogenic milling can be used to reduce the feed stream to fine powder. This allows sufficient dispersion in the extraction solvent.
  • cryogenic milling to disperse Feed Stream B makes it the least preferred feed stream of the three streams that were trialled, although as noted this problem could be overcome by dispersing the feed stream in a small amount of solvent first.
  • the current belief is that the "gumming up” is caused by the phosphatidyl choline (PC) not being solublised sufficiently and forming a gel.
  • PC phosphatidyl choline
  • Feed Stream A is preferred because it contains a much higher proportion of gangliosides which means that the residue (produced using 30:1 solvent to feed stream ratio) contains both a reasonable level of PS (32.7% PS) and a ganglioside content of around 6%.
  • PS 32.7% PS
  • ganglioside content of around 6%.
  • the residue produced using Feed Stream C contained a higher level of phospholipids, it is desirable for many health applications to produce a product enriched in both PS and gangliosides making Feed Stream A the preferred stream of the two.
  • the co-product (“extract”) from Feed Stream A (produced using 30:1 solvent to feed stream ratio) contained 10.2% SM. However this product was also high in neutral lipids and contained only 37.5% phospholipids. The product could be processed further to remove the neutral lipids using supercritical CO 2 , degumming or acetone.
  • Feed stream C contains fewer impurities than Feed Stream A. This highlights the fact that reducing the lactose and ash content of the feed stream upfront will have a very positive effect on the purity of the PS extract that can be attained.
  • Feed Stream B was the least desirable stream, because of its gummy nature, the residue was similar to the residue from extraction using Feed Stream A, having similar levels of PS (23.0%) and gangliosides (6.0%). And the co-product ("extract") is more attractive, because it contains very low levels of neutral lipids, high levels of phospholipids (63.7%) and has 14.1% SM.
  • Feed Stream A is the preferred stream for the process described in Example 1, the yield is of concern, as only 9% of the total feed stream solids is converted to a PS enriched product. This meant the throughput of the process is low, and 91% of the feed stream solids ended up as the co-product (the "extract").
  • Example 2 differs from Example 1 in that many co-products ("extracts") were produced which could be kept separately.
  • the process achieved complete fractionation of Feed Stream B into fractions containing high sphingomyelin and phosphatidyl serine.
  • Example 1 it was found in Example 1 that the solvent ratio of 30: 1 with Feed Stream A yields a 32.7% PS fraction. With a 50:1 solvent to feed ratio the fraction is 38.9% PS.
  • the 30:1 solvent ratio also has advantages however. This lower solvent ratio allows for smaller extraction vessels at full scale so will minimise cost and improve throughput.
  • This example shows the effect of water content on PS enrichment at 65°C.
  • Example 5 Effect of sample to solvent ratio on ethanol fractionation of Dairy Lipid extract (Feed Stream B)
  • This example shows the effect of sample to solvent ratio on PS enrichment at 65°C.
  • This example shows the effect of pH on PS enrichment at 65°C.
  • Dairy lipid extract (0.8g) (Feed Stream B) was added to 40ml of 95% ethanol at 65°C.
  • the ethanol contained either 5% water, 5% glacial acetic acid, or 5% aqueous ammonia (2.5%), and the pH of the three mixtures were 7.7, 3.2 and 10.0 respectively.
  • the mixtures were agitated by periodic shaking over 2 hours, then filtered.
  • the fractions were dried under vacuum and the analysed by 31 P NMR. The results are shown in Table 7 below. Acidic conditions resulted in the best PS enrichment, but the results were similar across the wide pH range.
  • This example shows the effect of the alcohol chain length on PS enrichment.
  • Example 8 Ethanol Fractionation of Phospholipids from various sources (sample to solvent 1:10)
  • This example shows that the process of the invention is broadly applicable to a wide range of lipid substrates using a sample to solvent ratio of 1 : 10 and a temperature of 50°C. This example also shows that cardiolipin is substantially concentrated by a process of the invention when present in the feed material.
  • Phospholipid rich extract (0.5g) from the following sources:
  • test tubes were weighed in test tubes and 5g of 95% ethanol added.
  • the test tube was stoppered and placed in a water bath at 50 0 C, with periodic shaking and ultrasonication over a 2 hour period.
  • the test tubes were removed and centrifuged (3000 rpm, 2min) and the supernatant decanted from the solid.
  • Solvent was removed from the fractions under a stream of argon and then under vacuum.
  • Phospholipid profiles were determined by 31 P NMR. The results are shown in Table 9.
  • This example shows that PS can be enriched from feed materials in which the level of PS is low.
  • the present invention has utility in providing products with high levels of particular phospholipids/sphingolipids including phosphatidyl serine, gangliosides and sphingomyelin. These products may be employed in a number of applications, including infant formulas, brain health, sports nutrition, drug delivery, dermatological applications, and arthritis.
  • phospholipids/sphingolipids including phosphatidyl serine, gangliosides and sphingomyelin.

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  • Life Sciences & Earth Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Nutrition Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Health & Medical Sciences (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Fats And Perfumes (AREA)
  • Fodder In General (AREA)

Abstract

La présente invention concerne des procédés permettant de fractionner une matière d'alimentation en composants solubles et insolubles par la mise en contact d'une matière d'alimentation et d'un solvant et, par la suite, par la séparation du solvant contenant les composants solubles des composants insolubles, la matière d'alimentation comprenant un ou plusieurs des éléments suivants : au moins 1 % p/p de phosphatidyl sérine, au moins 1 % p/p de sphingomyéline, au moins 1 % p/P des cardiolipine, ou au moins 0,3 % p/p de gangliosides, et le solvant comprenant un ou plusieurs alcools monovalents en C1-C3 et de l'eau (la teneur en eau du ou des alcools étant de 0 à 40 % v/v).
PCT/NZ2007/000088 2006-04-20 2007-04-20 PROCÉDÉ PERMETTANT DE SÉPARER des MATIÈRES lipidiques Ceased WO2007123425A1 (fr)

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009051502A1 (fr) * 2007-10-19 2009-04-23 Steven Charles Hodgkinson Procédés permettant de maintenir ou de stimuler la croissance ou le développement cognitif
EP3555254A2 (fr) * 2016-12-15 2019-10-23 FrieslandCampina Nederland B.V. Fractionnement bidimensionnel de la matière grasse du lait
CN111929133A (zh) * 2011-11-30 2020-11-13 新加坡科技研究局 用于生物学检测的gm1神经节苷脂与膜联蛋白v的微粒多肽比例
US12465619B2 (en) 2018-06-22 2025-11-11 Fonterra Co-Operative Group Limited Use of polar lipids to treat or prevent gestational diabetes mellitus

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5532141A (en) * 1995-06-13 1996-07-02 Holler; Larry D. Process for obtaining ganglioside lipids
JP2002241385A (ja) * 2001-02-15 2002-08-28 Yakult Honsha Co Ltd ホスファチジルセリンの分画法
WO2004047554A1 (fr) * 2002-11-26 2004-06-10 Phares Pharmaceutical Research N.V. Compositions de lipides marins
US20050170063A1 (en) * 2004-01-29 2005-08-04 Lalit Chordia Production of powder and viscous material

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5532141A (en) * 1995-06-13 1996-07-02 Holler; Larry D. Process for obtaining ganglioside lipids
JP2002241385A (ja) * 2001-02-15 2002-08-28 Yakult Honsha Co Ltd ホスファチジルセリンの分画法
WO2004047554A1 (fr) * 2002-11-26 2004-06-10 Phares Pharmaceutical Research N.V. Compositions de lipides marins
US20050170063A1 (en) * 2004-01-29 2005-08-04 Lalit Chordia Production of powder and viscous material

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PATENT ABSTRACTS OF JAPAN *

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009051502A1 (fr) * 2007-10-19 2009-04-23 Steven Charles Hodgkinson Procédés permettant de maintenir ou de stimuler la croissance ou le développement cognitif
AU2008312069B2 (en) * 2007-10-19 2014-03-06 Fonterra Co-Operative Group Limited Methods of maintaining or increasing growth or cognitive development
EP2211629A4 (fr) * 2007-10-19 2014-08-20 Fonterra Co Operative Group Procédés permettant de maintenir ou de stimuler la croissance ou le développement cognitif
EP2211629B1 (fr) 2007-10-19 2020-07-08 Fonterra Co-Operative Group Limited Procédés permettant de maintenir ou de stimuler la croissance ou le développement cognitif
EP3763219A1 (fr) 2007-10-19 2021-01-13 Fonterra Co-Operative Group Limited Procédés permettant de maintenir ou de stimuler la croissance ou le développement cognitif
CN111929133A (zh) * 2011-11-30 2020-11-13 新加坡科技研究局 用于生物学检测的gm1神经节苷脂与膜联蛋白v的微粒多肽比例
EP3555254A2 (fr) * 2016-12-15 2019-10-23 FrieslandCampina Nederland B.V. Fractionnement bidimensionnel de la matière grasse du lait
US12465619B2 (en) 2018-06-22 2025-11-11 Fonterra Co-Operative Group Limited Use of polar lipids to treat or prevent gestational diabetes mellitus

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