Prevention of MCPD formation by adjunctive degumming

Abstract

A method for preventing or reducing the formation of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) in triacylglyceride oils is provided.

Description

[Technical field]

[0001] The present invention relates to oil refining, and in particular to improved degumming and mechanical refining of triacylglyceride oils to reduce or completely remove monochloropropanediol esters (MCPDEs) from the refined oil.

[0002] [Background technology] 3-Halogen-1,2-propanediols, especially 3-monochloro-1,2-propanediol (3-MCPD), are known contaminants in food (Food Addit. Contam. (2006) 23:1290-1298). For example, studies have shown that 3-MCPD can be carcinogenic to rats when administered in high doses (Evaluation of Certain Food Additives and Contaminants, World Health Organisation, Geneva, Switzerland (1993) 267-285; lnt. J. Toxicol. (1998) 17:47).

[0003] 3-MCPD was originally discovered in acid hydrolyzed vegetable proteins (acid-HVP; Z. Lebensm.-Unters. Forsch. (1978) 167:241-244). More recently, refined edible oils have been found to contain 3-MCPD in the fatty acid ester form, but in very small amounts of free 3-MCPD (Food Addit. Contam. (2006) 23:1290-1298). The European Food Safety Authority (EFSA) recommends that 3-MCPD esters be treated equivalently to free 3-MCPD from a toxicity standpoint (European Food Safety Authority (2008)).

[0004] It has been reported that chlorination of acylglycerides can occur at very high temperatures, for example during the final step of the oil refining process, i.e., deodorization, where the oil can be heated up to 260-270 °C under vacuum (3-7 mbar). This can result in the formation of fatty acid esters of MCPD.

[0005] Limited options exist for efficient reduction of MCPD esters, posing a challenge to the vegetable oil refining industry. Currently, the presence of 3-MCPD in refined oils is carefully monitored, and oils with 3-MCPD content above a certain threshold are discarded to ensure full compliance with EFSA recommendations.

[0006] Because 3-MCPD can occur in many commercially important refined oils, such as vegetable oils, there is a particular need for improved methods of removing and / or avoiding such impurities that arise during oil refining.

[0007] [Summary of the Invention] The present inventors have developed a method that can substantially reduce or prevent the formation of MCPD and MCPD esters (MCPDE, including mono- and diesters) during the oil refining process.

[0008] The principle of this method is that phospholipid and / or wax-free oils (e.g. palm) can be refined using a combined refining process with the refining action of the phospholipid and wax components present in a particular crude oil. This method takes advantage of the high phospholipid and / or wax content of the crude oils or their gum extracts to improve the refining of an otherwise low phospholipid oil.

[0009] As a result, insoluble and soluble chlorine or chloride-containing substances that may serve as chlorine sources are concentrated in the aqueous and sediment fractions of the oil and can therefore be separated from the oil being refined. Separation can occur by mechanical treatments such as centrifugation, sedimentation, filtration, or traditional degumming or other refining processes. The method of the present invention can be applied to crude or partially refined (e.g., centrifuged, degummed, or bleached) triacylglycerol (also called triacylglyceride) oils, including but not limited to palm oil, palm stearin, palm olein, and their various fractions, palm kernel oil, coconut oil, sunflower oil, high oleic sunflower oil, and variants thereof, canola / rapeseed oil, corn oil, soybean oil, fish oil, algae oil, oils obtained from yeast, oils obtained from fungi, cocoa butter, and any mixtures or blends thereof.

[0010] Mechanical treatment may include centrifugation and / or sedimentation, either before, during, or after any other refining, refining, or deodorizing steps.

[0011] The degumming process may include water degumming, acid degumming, a combination thereof, dry degumming, base degumming, chemical refining, or a combination thereof.

[0012] Once the potential source of chlorine is removed, it is no longer available for the formation of chlorinated compounds such as MCPD, MCPD monoesters, and MCPD diesters during the heating step of oil refining, thereby resulting in a product oil that is low in chlorine-bearing materials and that can be subjected to various refining procedures, such as thermal treatment and deodorization, with the goal of producing a refined oil that is reduced or free of MCPD and MCPDE. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 shows MCPDE concentrations. [Diagram 2] FIG. 2 shows MCPD concentrations. [Diagram 3]FIG. 3 is a diagram showing the ratio of MCPD as an accumulation factor. [Figure 4] FIG. 4 shows MCPDE concentrations. [Diagram 5] FIG. 5 shows accumulation of MCPD. [Figure 6] FIG. 6 shows MCPDE concentrations. [Figure 7] FIG. 7 shows the concentration of 3-MCPD. [Figure 8] FIG. 8 shows the relative abundance of 3-MCPD. [Figure 9] FIG. 9 shows the concentration of 3-MCPD. [Figure 10] FIG. 10 shows the concentration of 3-MCPD. [Figure 11] FIG. 11 shows the concentration of 3-MCPD. [Figure 12] FIG. 12 shows the concentration of 3-MCPD. [Figure 13] FIG. 13 shows the concentration of 3-MCPD. [Figure 14] FIG. 14 shows the concentration of 3-MCPD.

[0014] A further advantage of the method of the present invention is that it allows for the use of lower temperatures in the deodorization of oil, which is 1) Reducing the formation of trans fatty acids (the formation of trans fatty acids at high temperatures is reported in Baley's industrial oil and fat products; Sixth Edition; Volume 5 Edible Oil and Fat Products: Processing Technologies; Chapter 8 Deodorization; section 3. Refined oil quality, subsection 3.2 Fat isomerization and degradation products). 2) reduce the formation of glycidyl esters (see overview of GE removal methods in “Glycidyl fatty acid esters in refined edible oils: a review on formation, occurrence, analysis, and elimination methods” in Comprehensive Reviews in Food Science and Food Safety; vol. 16, 263-281; ​​2017).

[0015] Thus, in one aspect, the present invention provides a method for preventing or reducing the formation of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) in a triacylglyceride oil, comprising: (a) a triacylglyceride oil, 1. Auxiliary oils having a higher phospholipid content and / or wax content than triacylglyceride oils, and / or 2. Mixing with a gum extract from oil; (b) degumming the triacylglyceride oil blend and / or optionally crystallizing insoluble components; (c) optionally, mixing the insoluble and crystallizable components with 1. by subjecting the triacylglyceride oil mixture to centrifugal force; and / or 2. By allowing the insoluble and crystallized components to settle by gravity, concentrating the triacylglyceride oil blend; (d) separating the insoluble and crystallizable components from the triacylglyceride oil blend and / or optionally subjecting the blend to one or more processes selected from degumming, physical refining, chemical refining, neutralization, interesterification, bleaching, winterization, and fractionation; (e) applying a heat treatment to the triacylglyceride oil blend.

[0016] In some embodiments, the starting triacylglyceride oil is a vegetable oil, an animal oil, a fish oil, a yeast oil, a fungal oil or an algae oil, preferably a vegetable oil. Starting triacylglyceride oil refers to the triacylglyceride oil before it is mixed in step (a) of the present method.

[0017] In some embodiments, the source triacylglyceride oil is palm oil or a fraction derived from palm oil.

[0018] In some embodiments, the source triacylglyceride oil is a fish oil or a fraction obtained from a fish oil.

[0019] In some embodiments, the feed triacylglyceride oil is a crude oil.

[0020] In some embodiments, the starting triacylglyceride oil is a partially refined oil or oil mixture that has been refined by any of centrifugation, sedimentation, filtration, washing, winterization, fractionation, degumming, bleaching, or deodorization, or any combination thereof.

[0021] In one embodiment, the raw triacylglyceride oil is water degummed without the use of acid, which can be phosphoric acid, citric acid, sulfuric acid, formic acid, or acetic acid.

[0022] In some embodiments, the source triacylglyceride oil is a mixture of crude and partially refined oil.

[0023] In one embodiment, the source triacylglyceride oil is bleached.

[0024] In one embodiment, the feed triacylglyceride oil is contacted with a bleaching earth.

[0025] In one embodiment, the feed triacylglyceride oil is mixed with bleaching earth.

[0026] In one embodiment, the starting triacylglyceride oil has been bleached with at least 0.01 wt. %, preferably at least 0.1 wt. %, more preferably at least 0.5 wt. % bleaching earth.

[0027] In one embodiment, the feed triacylglyceride oil is contacted with at least 0.01 wt. %, preferably at least 0.1 wt. %, more preferably at least 0.5 wt. % bleaching earth.

[0028] In one embodiment, the starting triacylglyceride oil is mixed with at least 0.01% by weight, preferably at least 0.1% by weight, more preferably at least 0.5% by weight of bleaching earth.

[0029] In one embodiment, the raw triacylglyceride oil is mixed with bleaching earth and the bleaching earth is removed from the oil.

[0030] In one embodiment, the raw triacylglyceride oil is mixed with bleaching earth and the bleaching earth is not removed from the oil prior to step a).

[0031] In one embodiment, the combining in step (a) comprises heating to a temperature above the melting temperature of the starting triacylglyceride oil and the auxiliary oil, and / or homogenizing the mixture.

[0032] In one embodiment, the combining in step (a) comprises incubating the starting triacylglyceride oil and the auxiliary oil at a temperature above the melting temperature of the starting triacylglyceride oil and the auxiliary oil, and / or homogenizing the mixture.

[0033] In one embodiment, the combining in step (a) comprises heating the starting triacylglyceride oil and the gum extract from the oil to a temperature above the melting temperature and / or homogenizing the mixture.

[0034] In one embodiment the gum extract of oil is heated to a temperature in excess of 100°C or 120°C or 140°C or 160°C or 180°C or 200°C prior to the combining step (a).

[0035] In one embodiment, the auxiliary oil is heated to a temperature greater than 100°C, or 120°C, or 140°C, or 160°C, or 180°C, or 200°C prior to blending step (a).

[0036] In one embodiment, the admixture of step (a) is heated to a temperature greater than 100° C., or 120° C., or 140° C., or 160° C., or 180° C., or 200° C. prior to step (b).

[0037] In one embodiment, the gum extract of oil, or auxiliary oil, or admixture of step a) is incubated for at least 1 minute.

[0038] In one embodiment, the admixture of step (a) is heated in a closed vessel under a pressure higher than 1 bar or 3 bar or 5 bar prior to step (b).

[0039] In one embodiment, in step (b), the admixture is cooled to a temperature of 20° C., or 10° C., or 5° C., or 4° C., or 0° C., or sub-zero to facilitate the crystallization process and precipitation of the insoluble components.

[0040] In one embodiment, the gum extract of oil does not contain any added acid.

[0041] In another embodiment, the degumming process of step (b) is carried out by adding no acid to the oil and by adding water that does not contain any added acid.

[0042] In one embodiment, the oil gum extract is fractionated prior to use.

[0043] In another embodiment, the gum extract of oil is purified prior to use.

[0044] In one embodiment, the combining in step (a) comprises incubating the starting triacylglyceride oil and the gum extract obtained from the oil at a temperature above the melting temperature of the starting triacylglyceride oil and the gums, and / or homogenizing the mixture.

[0045] In one embodiment, the temperature of the blend in step (a) is adjusted to at least 10° C. higher than the melting point of the starting triacylglyceride oil or at least 10° C. higher than the melting point of the auxiliary oil, whichever is higher, and the temperature is preferably adjusted from step (a) to step (d).

[0046] In one embodiment, the temperature of the blend in step (a) is adjusted to at least 20° C. higher than the melting point of the starting triacylglyceride oil or at least 20° C. higher than the melting point of the auxiliary oil, whichever is higher, and the temperature is preferably adjusted from step (a) to step (d).

[0047] In one embodiment, the temperature of the blend in step (a) is adjusted to at least 30° C. above the melting point of the starting triacylglyceride oil or at least 30° C. above the melting point of the auxiliary oil, whichever is higher, and the temperature is preferably adjusted from step (a) to step (d).

[0048] In one embodiment, the temperature of the blend in step (d) is adjusted to at least 10° C. above the melting point of the starting triacylglyceride oil or at least 10° C. above the melting point of the auxiliary oil, whichever is greater.

[0049] In one embodiment, the temperature of the blend in step (d) is adjusted to at least 20° C. above the melting point of the starting triacylglyceride oil or at least 20° C. above the melting point of the auxiliary oil, whichever is greater.

[0050] In one embodiment, the temperature of the blend in step (d) is adjusted to at least 30° C. above the melting point of the starting triacylglyceride oil or at least 30° C. above the melting point of the auxiliary oil, whichever is greater.

[0051] In some embodiments, the insoluble and crystallized components are separated from the triacylglyceride oil blend by one or more of filtration, decantation, centrifugation, pumping, and draining.

[0052] In some embodiments, the gum extract of oil is obtained by one or more of precipitation, crystallization and sedimentation, and centrifugation.

[0053] In some embodiments, the gum extract of oil is obtained by one or more of water degumming and acid degumming.

[0054] In some embodiments, the gum extract of the oil is obtained by water-acid degumming.

[0055] In some embodiments, the gum extract of the oil is obtained by water degumming using water that does not contain any added acid.

[0056] In some embodiments, the gum extract of the oil is obtained by one or more of water degumming, acid degumming, acid degumming, water-acid degumming, super degumming, TOP degumming, UF degumming, organic refining, dry degumming, caustic refining, precipitation, crystallization and sedimentation, and centrifugation.

[0057] In some embodiments, insoluble components include, for example, particulates, separated droplets, emulsions, suspensions, and precipitates.

[0058] In another embodiment, the thermal treatment is deodorization (eg, by steam distillation or short path distillation).

[0059] In another embodiment, the heat treatment is carried out in a closed vessel.

[0060] In one embodiment, the heat treatment step removes unwanted components, which may be color pigments, free fatty acids, monoglycerides, trace contaminants and / or odors.

[0061] In one embodiment, the present invention provides a method for preventing or reducing the formation of monochloropropanediol (MCPD).

[0062] In one embodiment, the present invention provides a method for preventing or reducing the formation of monochloropropanediol esters (MCPDEs).

[0063] In one embodiment, step (a2) is performed and then step (a1) is performed.

[0064] In one embodiment, step (a1) is performed and then step (a2) is performed.

[0065] In one embodiment, applying a heat treatment comprises exposing the oil to a temperature in the range of 150-300° C., more preferably in the range of 160-290° C. or 160-240° C., preferably for at least 30 minutes.

[0066] In one embodiment, applying the heat treatment comprises exposing the oil to a temperature in the range of 140-200°C.

[0067] In one embodiment, the starting triacylglyceride oil is palm oil. In one embodiment, the starting triacylglyceride oil is palm oil and the heat treatment step comprises exposing the oil to a temperature in the range of 160-290°C.

[0068] In one embodiment, the starting triacylglyceride oil is a fish oil. In one embodiment, the starting triacylglyceride oil is a fish oil and the heat treatment step comprises exposing the oil to a temperature in the range of 140-220°C.

[0069] In one embodiment, the starting triacylglyceride oil is sunflower oil and the heat treatment step comprises exposing the oil to a temperature in the range of 160-240°C.

[0070] In one embodiment, the amount of monochloropropanediol (MCPD) or monochloropropanediol ester (MCPDE) is measured after the heat treatment step (e).

[0071] In one embodiment, the amount of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) is measured after the heat treatment step (e) and the amount of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) is reduced by at least 30% compared to a heat treated oil refined using the same protocol but without the use of an adjunct oil or gum extract.

[0072] In one embodiment, the amount of MCPDE in the thermally treated oil of step (e) is reduced by at least a factor of two, as measured by direct LC-MS.

[0073] In one embodiment, the feed triacylglyceride oil in step (a) is a crude triacylglyceride oil.

[0074] In one embodiment, the starting triacylglyceride oil is not degummed prior to step (a).

[0075] In one embodiment, the starting triacylglyceride oil is degummed prior to step (a).

[0076] In one embodiment, the starting triacylglyceride oil is not decolorized prior to step (a).In one embodiment, the starting triacylglyceride oil is not fractionated prior to step (a).

[0077] In one embodiment, the starting triacylglyceride oil is degummed prior to step (a). In one embodiment, the starting triacylglyceride oil is decolorized prior to step (a). In one embodiment, the starting triacylglyceride oil is fractionated prior to step (a).

[0078] In one embodiment, the starting triacylglyceride oil is neutralized prior to step (a).

[0079] In one embodiment, the starting triacylglyceride oil is not neutralized prior to step (a).

[0080] In a preferred embodiment, the starting triacylglyceride oil is not deodorized prior to step (a).

[0081] In one embodiment, the starting triacylglyceride oil is subjected to pre-washing prior to step (a). In one embodiment, the starting triacylglyceride oil is subjected to pre-refining prior to step (a). In one embodiment, the starting triacylglyceride oil is subjected to hydrogenation prior to step (a). In one embodiment, the starting triacylglyceride oil is subjected to interesterification prior to step (a).

[0082] In one embodiment, the source triacylglyceride oil is a vegetable oil, an animal oil, a fish oil, or an algae oil.

[0083] In one embodiment, the starting triacylglyceride oil is crude palm oil and the process is applied starting from step (a).

[0084] In one embodiment, the starting triacylglyceride oil is fractionated crude palm oil and the process is applied starting from step (a).

[0085] In one embodiment, the starting triacylglyceride oil is crude palm kernel oil and the process is applied starting from step (a).

[0086] In one embodiment, the starting triacylglyceride oil is fractionated crude palm kernel oil and the process is applied starting from step (a).

[0087] In one embodiment, the starting triacylglyceride oil is crude coconut oil and the method is applied starting from step (a).

[0088] In one embodiment, the starting triacylglyceride oil is fractionated crude coconut oil and the method is applied starting from step (a).

[0089] In one embodiment, the starting triacylglyceride oil is an oil obtained from algae, or yeast, or fungi, and the method is applied starting from step (a).

[0090] In one embodiment, the source triacylglyceride oil is crude fish oil.

[0091] In one embodiment, the source triacylglyceride oil is a crude algal oil.

[0092] In one embodiment, the source triacylglyceride oil is a crude fungal oil.

[0093] In one embodiment, the source triacylglyceride oil is a crude yeast oil.

[0094] In one embodiment, the starting triacylglyceride oil is a crude seed oil and the method is applied starting from step (a). For example, the crude seed oil can be sunflower oil, canola / rapeseed oil, corn oil.

[0095] In a preferred embodiment, the starting triacylglyceride oil is a vegetable oil, preferably the vegetable oil is selected from the group consisting of palm oil, sunflower oil, corn oil, canola oil, soybean oil, corn oil, coconut oil, palm kernel oil, and cocoa butter, hi one embodiment, the starting triacylglyceride oil is palm oil.

[0096] In one embodiment, the adjunct oil is crude sunflower oil or a high oleic variant thereof.

[0097] In one embodiment, the adjunct oil is crude soybean oil or a high oleic variant thereof.

[0098] In one embodiment, the adjunct oil is crude rapeseed oil or a high oleic variant thereof.

[0099] In one embodiment the gum extract is obtained from crude rapeseed oil or a high oleic variant thereof.

[0100] In one embodiment, the gum extract is obtained from crude soybean oil or a high oleic variant thereof.

[0101] In one embodiment, the gum extract is obtained from crude sunflower oil or a high oleic variant thereof.

[0102] In one embodiment, the adjunct oil generates 10 times less, or 3 times less, or 2 times less MCPD upon heating than the starting triacylglyceride oil.

[0103] In one embodiment, the heat treated auxiliary oil generates less than 10 ppm, or less than 3 ppm, or less than 1 ppm, or less than 0.5 ppm, or less than 0.2 ppm MCPD. In one embodiment, the starting triacylglyceride oil has a free fatty acid content of 0.5-25% (w / w%), or a free fatty acid content of 1-12% (w / w%), or a free fatty acid content of 3-7% (w / w%).

[0104] In another embodiment, the starting triacylglyceride oil has a free fatty acid content of at least 0.5 wt%, preferably 1 wt%, more preferably 3 wt%. In another embodiment, the starting triacylglyceride oil has a free fatty acid content of less than 25% (w / w%), preferably less than 15% (w / w%), more preferably less than 10% (w / w%).

[0105] In one embodiment, the starting triacylglyceride oil is not mixed with any alkali, such as sodium hydroxide or potassium hydroxide, or with any products that contain sodium hydroxide or potassium hydroxide, such as caustic soda, caustic potash, etc. In another embodiment, the starting triacylglyceride oil is not mixed with any ammonium hydroxide or any ammonium salts.

[0106] In one embodiment, the starting triacylglyceride oil is not mixed with salts, such as sodium, potassium, or ammonium salts. Examples of sodium salts include sodium chloride, sodium hypochlorite, sodium carbonate, sodium formate, sodium citrate, and sodium phosphate.

[0107] In another embodiment, the starting triacylglyceride oil has a soap content of less than 1000 ppm. In another embodiment, the starting triacylglyceride oil has a soap content of less than 20 ppm. In another embodiment, the starting triacylglyceride oil is soap-free.

[0108] In one embodiment, the raw triacylglyceride oil has not been acidified or subjected to acid degumming.

[0109] In another embodiment, the starting triacylglyceride oil is not mixed with acids less than 195 Da. In a preferred embodiment, the starting triacylglyceride oil is not mixed with acids less than 195 Da that have an anhydrous form.

[0110] In another embodiment, the starting triacylglyceride oil does not contain more than 0.01% of acids less than 195 Da. In another embodiment, the starting triacylglyceride oil does not contain more than 0.01% of acids having an anhydrous form less than 195 Da.

[0111] In another embodiment, the starting triacylglyceride oil does not contain more than 0.01% of acids having a logP<1. In another embodiment, the starting triacylglyceride oil does not contain more than 0.01% of acids having an acidity pKa1<5.

[0112] In another embodiment, the raw triacylglyceride oil is substantially free of any one of phosphoric acid, citric acid, sodium hydroxide, potassium hydroxide, boric acid, hypochloric acid, and hydrochloric acid. As used herein, sodium hydroxide may refer to caustic soda or alkali, and potassium hydroxide may refer to alkali potash.

[0113] In another embodiment, the starting triacylglyceride oil is substantially free of any one of phosphoric acid, citric acid, sodium chloride, sodium carbonate, sodium hydroxide, potassium hydroxide, phosphates, polyphosphates, acetic acid, acetic anhydride, calcium sulfate, calcium carbonate, sodium sulfate, boric acid, hypochlorous acid, hydrochloric acid, and tannic acid.

[0114] In another embodiment, the starting triacylglyceride oil is substantially free of any added ionic, cationic, and anionic surfactants, hi another embodiment, the starting triacylglyceride oil is substantially free of any emulsifiers, such as sorbitan esters or polyglycerol esters.

[0115] In another embodiment, the raw triacylglyceride oil is substantially free of any additives listed in Bailey's Industrial Oil and Fat Products-6th edition, page 2236 in Chapter Emulsifiers for the food industry-Table 4, page 262, such as sucrose, glycol, propylene glycol, and / or lactylates.

[0116] In one embodiment, the raw triacylglyceride oil has not been subjected to water degumming or wet degumming.

[0117] In another embodiment, the starting triacylglyceride oil has a moisture content of less than 1%, or less than 0.5%, or less than 0.3%, and in one embodiment, the starting triacylglyceride oil has a moisture content of less than 1%, or less than 0.3%, or less than 0.1%.

[0118] In one embodiment, the source triacylglyceride oil is a neutralized fish oil.

[0119] In one embodiment, the source triacylglyceride oil is a neutralized algal oil.

[0120] In one embodiment, the source triacylglyceride oil is a neutralized fungal oil.

[0121] In one embodiment, the source triacylglyceride oil is a neutralized yeast oil.

[0122] In a preferred embodiment, the starting triacylglyceride oil is completely immiscible with water and has a water content of less than 0.5%.

[0123] In another embodiment, the starting triacylglyceride oil does not contain added water.

[0124] In one embodiment, the starting triacylglyceride oil has a bleaching clay content of less than 0.01%. In another embodiment, the starting triacylglyceride oil is not mixed with bleaching clay. In another embodiment, the starting triacylglyceride oil is free of bleaching clay.

[0125] In one embodiment, the starting triacylglyceride oil is not bleached. In another embodiment, the starting triacylglyceride oil is not degummed. In another embodiment, the starting triacylglyceride oil is not neutralized.

[0126] In another embodiment, the starting triacylglyceride oil does not contain added crystallization agents, such as solvents. Such solvents may include hexane, acetone, and detergents as described in [The Lipid Handbook-Third Edition; edited by Frank D. Gunstone; Chapter 4.4.2.] and [Bailey's Industrial Oil and Fat Products-6th edition, Chapter 12] or sorbitan esters or polyglycerol fatty acid esters as described in [Omar et al Journal of Oil Palm Research Vol.27(2)June 2015 p.97-106]. The starting triacylglyceride oil may be crude palm oil.

[0127] In another embodiment, the starting triacylglyceride oil is not winterized.

[0128] In another embodiment, the starting triacylglyceride oil does not contain added substances, such as degumming agents, neutralizing agents, additives, solvents, salts, seeding agents, acids, bases, or buffers.

[0129] In another embodiment, the starting triacylglyceride oil is crude palm oil and does not contain added substances such as degumming agents, neutralizing agents, additives, solvents, salts, seeding agents, acids, bases, or buffers.

[0130] In one embodiment, the degumming step (b) uses any one of phosphoric acid, citric acid, sodium chloride, sodium carbonate, sodium hydroxide, potassium hydroxide, phosphates, polyphosphates, acetic acid, acetic anhydride, calcium sulfate, calcium carbonate, sodium sulfate, boric acid, hypochlorous acid, hydrochloric acid, and tannic acid.

[0131] In one embodiment, the degumming step (b) uses water.

[0132] In one embodiment, the degumming step (b) uses water in combination with any one of phosphoric acid, citric acid, sodium chloride, sodium carbonate, sodium hydroxide, potassium hydroxide, phosphates, polyphosphates, acetic acid, acetic anhydride, calcium sulfate, calcium carbonate, sodium sulfate, boric acid, hypochlorous acid, hydrochloric acid, and tannic acid.

[0133] In one embodiment, the raw triacylglyceride oil is pre-purified from insoluble material by centrifugation.

[0134] In one embodiment, the starting triacylglyceride oil has a crystallized triacylglycerol content of less than 10% (w / w%). In another embodiment, the starting triacylglyceride oil has a crystallized triacylglycerol content of less than 5% (w / w%). In one embodiment, the starting triacylglyceride oil has a crystallized triacylglycerol content of less than 2% (w / w%). In one embodiment, the starting triacylglyceride oil has a crystallized triacylglycerol content of less than 0.5% (w / w%).

[0135] As used herein, crystallized triacylglycerol refers to solid triacylglycerol or the solid portion of fat. The solid fat content of fats and oils can be determined by pulsed nuclear magnetic resonance [Bailey's Industrial Oil and Fat Products-6th edition, page 175 Chapter 5.2.1.].

[0136] In another embodiment, the starting triacylglyceride oil has not been cooled below 20°C, 15°C, or 10°C.

[0137] In one embodiment, the method further comprises one or more of the following steps after step (e): (f) one or more processes selected from the group consisting of physical or chemical refining, degumming, neutralization, and decolorization; (g) optionally deodorizing the product of step (f), preferably the deodorization being vacuum steam deodorization; and (h) optionally, fractionating the product of steps (f) and (g).

[0138] In another aspect, there is provided a refined triacylglyceride oil obtainable by the process of the present invention.

[0139] In one embodiment, the amount of monochloropropanediol esters (MCPDEs) in the heat-treated refined oil is reduced by a factor of two compared to heat-treated oil refined via the same protocol but without the use of auxiliary oils or their gums.

[0140] In one embodiment, the amount of monochloropropanediol esters (MCPDEs) in the heat-treated refined oil is at least 30% lower compared to a heat-treated oil refined via the same protocol but without the use of auxiliary oils or gums thereof.

[0141] In one embodiment, the amount of monochloropropanediol esters (MCPDEs) in the heat-treated refined oil is at least 50% lower compared to a heat-treated oil refined via the same protocol but without the auxiliary oil or gums thereof.

[0142] There is also provided a refined triacylglyceride oil according to the present invention for use in making a food product.

[0143] Food products produced by using refined triacylglyceride oils according to the present invention are also provided.

[0144] [Mode for carrying out the invention] As used herein, the terms "comprising," "comprises," and "comprised of" are synonymous with "including" or "includes" and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or steps. The terms "comprising," "comprises," and "comprised of" also include the terms "consisting of," "containing," or "contains."

[0145] mixing As used herein, the term "blending" refers to the process of mixing two or more components together in a vessel (e.g., a vessel, reservoir, tube, tank, container, vial, or pot) to enhance the homogeneity of the resulting blend. Mixing can include one or a combination of several different common practices including agitation, stirring, sonication, mixing, high shear mixing, and / or shaking, optionally accompanied by heating, for example, to enhance solubilization of the components and / or melt and ensure that both components are liquid.

[0146] purification Refining is particularly suitable for removing from the raw triacylglyceride oil insoluble fractions of oil that may contain chlorine / chloride-bearing contaminants (substances that may serve as the chlorine source required for the formation of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE)). Raw triacylglyceride oil as used throughout this specification is taken to mean the triacylglyceride oil immediately before being subjected to step (a) or step (h) of the process of the present invention.

[0147] The method of the present invention involves subjecting a raw triacylglyceride oil to a treatment that physically removes from the raw oil (e.g., crude oil) insoluble, soluble and precipitated fractions of oil containing chloride / chlorine-bearing substances that can be active sources of chlorine during oil refining. The treatment is based on a combined refining process that utilizes the refining action of phospholipid and wax components present in certain crude oils to purify these oils that are low in phospholipids and / or wax. In this method, the high phospholipid and / or wax content of the crude oil or its gum extract is utilized to improve the refining of another low phospholipid oil.

[0148] As a result, insoluble, precipitated, crystalline, and water-soluble chlorine or chloride-containing materials that can serve as chlorine sources are concentrated in the aqueous, gum, and precipitate fractions of the oil and can therefore be separated from the oil being refined. Separation can occur by mechanical treatments such as centrifugation, sedimentation, filtration, or traditional degumming or other refining processes. The method of the present invention can be applied to crude or partially refined triacylglycerol (also called triacylglyceride) oils, including but not limited to palm oil, palm stearin, palm olein, and their various fractions, palm kernel oil, coconut oil, sunflower oil, corn oil, high oleic sunflower oil, and variants thereof, canola / rapeseed oil, soybean oil, fish oil, algae oil, oils obtained from yeast, oils obtained from fungi, cocoa butter, and any mixture or blend thereof.

[0149] Separation of the insoluble and crystallized components from the triacylglyceride oil mixture can occur via filtration and / or decantation and / or centrifugation and / or pumping and / or drainage.

[0150] The degumming process may include water degumming, acid degumming, a combination thereof, dry degumming, base degumming, chemical refining, or a combination thereof.

[0151] 3-Halogen-1,2-propanediols, especially 3-monochloro-1,2-propanediol (3-MCPD), are known contaminants in food (Food Addit. Contam. (2006) 23:1290-1298). For example, studies have shown that 3-MCPD can be carcinogenic to rats when administered in high doses (Evaluation of Certain Food Additives and Contaminants, World Health Organisation, Geneva, Switzerland (1993) 267-285; lnt. J. Toxicol. (1998) 17:47). However, it has also been discovered that refined edible oils, while they may contain 3-MCPD in its fatty acid ester form, may contain only very small amounts of free 3-MCPD (Food Addit. Contam. (2006) 23:1290-1298). The European Food Safety Authority (EFSA) recommends that, from a toxicity standpoint, 3-MCPD esters be treated equally to free 3-MCPD (European Food Safety Authority (2008)).

[0152] It is well known that dehalogenation reactions can occur during thermal processes. It has been shown that when sufficient activation energy is input, which is abundant during deodorization of vegetable oils at high temperatures (e.g., up to 270°C), chlorine remains in its chemical form as hydrogen chloride (gas). The inventors believe that hydrogen chloride can be generated during oil refining from chlorine-containing compounds that are originally present in the starting material, e.g., vegetable material, of the triacylglyceride oil refining process.

[0153] In fact, it has been suggested that the MCPD formation reaction increases exponentially (above 150°C) and is completed in a short time.

[0154] Without wishing to be bound by theory, it is suggested mechanistically that MCPD diesters may be formed during oil refining via protonation of the terminal ester groups of triacylglycerides (TAGs), which are present in approximately 88%-95% of the total glycerides in most vegetable oils, i.e., by interaction with hydrogen chloride generated during oil refining. The oxonium cations formed can then undergo intramolecular rearrangement followed by nucleophilic displacement of chloride ions to release free fatty acids and MCPD diesters.

[0155] Once removed through the use of the method of the present invention, the potential chlorine source is no longer available for the formation of chlorinated compounds such as MCPD esters during the heating step of oil refining, thereby resulting in a refined product oil that, when subjected to various refining steps involving thermal treatment, e.g., deodorization, has reduced generation of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) compared to unrefined refined triacylglyceride oil.

[0156] In another embodiment, the amount of monochloropropanediol esters (MCPDE) and MCPD is reduced by at least 30%, 50%, 60%, 70%, 80%, 90%, 95%, or 99% in the refined and thermally treated triacylglyceride oil compared to the refined raw triacylglyceride oil that does not contain auxiliary oils or gums thereof.

[0157] Refined oils produced using the methods of the present invention may contain, for example, less than 3 ppm, less than 1 ppm, less than 0.5 ppm, less than 0.3 ppm, or preferably less than 0.1 ppm MCPD.

[0158] The amount of MCPDE can be easily analyzed using protocols well known in the art. For example, as shown in this example, a liquid chromatography / mass spectrometry (LC / MS)-based approach is suitable for analyzing the concentration of MCPDE.

[0159] In one embodiment, the feed triacylglyceride oil input to step (a) of the process of the present invention is a crude triacylglyceride oil.

[0160] As used herein, the term "crude oil" may refer to unrefined oil. For example, in some embodiments, the raw triacylglyceride oil input to step (a) of the method of the present invention has not been refined, degummed, bleached, and / or fractionated. In a preferred embodiment, the raw triacylglyceride oil has not been deodorized before step (a).

[0161] In some embodiments, the starting triacylglyceride oil is subjected to pre-processing, such as pre-washing, prior to step (a). However, any process performed on the starting triacylglyceride oil prior to step (a) preferably does not involve heating the triacylglyceride oil to a temperature greater than 100° C., 150° C., 200° C., or 250° C. In some embodiments, the triacylglyceride oil is subjected to pre-refining, fractionation, hydrogenation, and / or transesterification prior to step (a).

[0162] Triacylglyceride Oil The term "triacylglyceride" can be used interchangeably with "triacylglycerol" and "triglyceride." In these compounds, each of the three hydroxyl groups of glycerol is esterified with a fatty acid. Oils that may be refined using the methods of the present invention include triacylglycerides, including vegetable oils, animal oils, fish oils, algae oils, and combinations thereof.

[0163] In a preferred embodiment, the starting triacylglyceride oil is a vegetable oil, such as sunflower oil, corn oil, canola oil, soybean oil, coconut oil, palm oil, palm kernel oil, and cocoa butter.

[0164] In another embodiment, the starting triacylglyceride oil is palm oil or a fractionated palm oil such as palm olein, palm stearin, or a mid fraction.

[0165] In a preferred embodiment, the source triacylglyceride oil is a crude vegetable oil.

[0166] In one embodiment, the source triacylglyceride oil is obtained from a unicellular organism.

[0167] In one embodiment, the source triacylglyceride oil is obtained from fish.

[0168] In one embodiment, the source triacylglyceride oil is obtained from algae.

[0169] In one embodiment, the source triacylglyceride oil is obtained from a fungus.

[0170] In one embodiment, the source triacylglyceride oil is obtained from yeast.

[0171] In another preferred embodiment, the starting triacylglyceride oil is crude palm oil or a fractionated crude palm oil such as crude palm olein, crude palm stearin, crude midpoint fraction.

[0172] In one embodiment, the vegetable oil is crude palm oil. In one embodiment, the vegetable oil is crude corn oil. In one embodiment, the vegetable oil is crude sunflower oil. In one embodiment, the vegetable oil is crude cold pressed canola oil. In one embodiment, the vegetable oil is crude soybean oil.

[0173] In a preferred embodiment, the vegetable oil is at least partially solvent extracted. Preferably, the solvent is n-hexane or a mixture of 2-propanol and n-hexane.

[0174] Crude triacylglyceride oil In the case of palm oil, crude oil can be produced from different parts of the palm fruit, for example from the flesh of the fruit known as the mesocarp, and also from the seed or kernel of the fruit. Extraction of crude palm oil (CPO) from the crushed fruit can be carried out, for example, at temperatures ranging from 90 to 140°C.

[0175] In other cases, for example, crude sunflower oil is produced by pressing, by solvent extraction, or by a combination thereof, as described, for example, in Gotor & Rhazi in Oilseeds&fats Crops and lipids 2016 (DOI:10.1051 / ocl / 2016007).

[0176] refined oil As used herein, the term "refined" may refer to oil that has been subjected to a process that improves the quality of the oil, including heat treatment. The heat treatment may be a deodorization process that includes steam distillation or short path distillation. Such heat treatment may be carried out at temperatures in the range of 150-300°C, more commonly in the range of 160-260°C or 160-240°C.

[0177] Gum As used herein, the term "gums" may refer to sludge, meal particle sediment impurities, crystallized waxes, sediments, glycolipids, sugars, and primarily phospholipids and phospholipid-based precipitates that vegetable oils release upon storage, cooling, or addition of acid and / or water. Gums may be removed from oils by one or more of water degumming, acid degumming, acid degumming, water-acid degumming, super degumming, TOP degumming, UF degumming, organic refining, dry degumming, caustic refining, precipitation, crystallization and sedimentation, and centrifugation [Chapter 6 Enzymatic degumming by Ch.Dayton&F.Galhardo in Green Vegetable Oil Processing].

[0178] Gum Extract As used herein, the term "gum extract" may refer to a gum obtained from either an oil or a fraction or component thereof.

[0179] lecithin As used herein, the term "lecithin" may refer to the water soluble fraction of a "gum." Thus, the term "gum" includes "lecithin" and "lysolecithin."

[0180] Heat Treatment As used herein, the term "thermal treatment" may refer to exposing the oil to temperatures in the range of 150-300° C., more commonly in the range of 160-260° C. or 160-240° C. Thermal treatment may be carried out in a closed vessel or ampoule, or in combination with vacuum and / or steam, as is done during industrial settings (steam distillation or short path distillation) during deodorization.

[0181] Chlorine and Chlorides Chlorine is a chemical element with the symbol Cl and atomic number 17. Chlorine can be found in a wide variety of substances in both ionic (e.g., sodium chloride) and covalent (e.g., polyvinyl chloride) forms. Thus, the terms "chlorine" and "chloride" both refer to substances that contain the element chlorine in various forms.

[0182] As used herein, the terms "chlorine-containing," "chloride-containing," "organochlorine," and "chlorine donor" all refer to substances that contain the element chlorine in any form, whether ionic, polar covalent, or covalent.

[0183] Chlorine or chloride-bearing substances As used herein, the term "chlorine or chloride bearing material" refers to a material that contains the element chlorine in any form, whether ionic, polar covalent, or covalent.

[0184] Chlorine Donor As used herein, the term "chlorine donor" refers to a substance that contains elemental chlorine in any form and can release chlorine in any form, such as, but not limited to, hydrochloric acid, hypochlorite, and chloride anion.

[0185] Acidity and pH In chemistry, pH is a measure used to describe how acidic or basic an aqueous solution is. Similarly, as used herein, the terms "pH" and "acidity" refer to the free acid content in an oil sample. For example, when an oil is mixed with phosphoric acid, its pH can be considered to be lowered. Similarly, the neutralization step of adding sodium hydroxide to an oil can be considered to increase the pH of the oil.

[0186] Melting Temperature As used herein, the term "melting temperature" may refer to the temperature at which a solid state changes from solid to liquid under a pressure of 100 kPa. For example, the melting temperature may be the temperature at which a solid state changes from solid to liquid when heated at 2°C / min under a pressure of 100 kPa.

[0187] One of skill in the art can readily select a suitable method for identifying the melting temperature of a triacylglyceride oil.

[0188] For example, an apparatus for melting temperature analysis may consist of a heating block or an oil bath with a transparent window (e.g., a Thiele tube) and a magnifying glass. A solid sample may be placed in a thin glass tube, which is placed in the heating block or immersed in an oil bath and then gradually heated. The melting of the solid can be observed and the associated melting temperature recorded.

[0189] For fats and oils containing highly complex triacylglycerol compositions, the slip melting point method is a commonly used measure (AOCS Official Method Cc3-25).

[0190] Centrifugation As used herein, the term "centrifugation" may refer to the rapid spinning of a container containing oil as its contents to apply centrifugal force to the container and its contents.

[0191] In one embodiment, the centrifugation is carried out at an elevated temperature where the oil is in a liquid state, which can be 30°C, 40°C, 50°C, 60°C, 70°C, 80°C, 100°C or more for palm oil, 50°C, 60°C, 80°C, 100°C or more for palm stearin, 15°C, 20°C or more for palm olein, and 5°C or more for seed oils including sunflower oil, canola oil / rapeseed oil, corn oil.

[0192] In a preferred embodiment, the temperature may be between 30° C. and 80° C., preferably between 35° C. and 70° C. for palm oil. In a preferred embodiment, the temperature may be between 5° C. and 20° C. for sunflower oil. In a preferred embodiment, the centrifugation speed is at least 15,000 g for 15 minutes.

[0193] Sedimentation As used herein, the term "settling" or "settling" refers to placing an oil container in a motionless or substantially motionless environment, preferably avoiding disturbance for a period of time which may be at least 4 hours, 6 hours, 1 day, 2 days, 1 week, or 1 month.

[0194] In one embodiment, the oil container is placed in a stationary, motionless environment, avoiding disturbance for a period of at least 5 months, for example, for crude sunflower oil or crude soybean oil, In one embodiment, the crude oil is heated to at least 60° C. prior to settling.

[0195] In one embodiment, the oil container is kept stationary in a stationary, motionless environment, avoiding disturbance for a period of at least 4 days, for example for cold-pressed crude canola oil.

[0196] Soap As used herein, the term "soap" can refer to a variety of cleaning and lubricating products made from materials with surfactant properties.

[0197] In the context of vegetable oil refining and in the present context, the term "soap" is used to denote alkali carboxylates, which are salts of fatty acids formed by negatively charged deprotonated fatty acids and positively charged counterions, such as sodium or potassium cations. [Bailey's Industrial Oil and Fat Products-6th edition, page 3084-Soap raw materials and their processing page 105; Wikipedia]

[0198] As is well known in the literature on alkaline refining practice, free fatty acids react with alkali, such as sodium hydroxide or potassium hydroxide, to form such soaps [The Lipid Handbook-Third Edition; edited by Frank D. Gunstone; pages 178, 191].

[0199] Further refinement Because the insoluble oil components along with their chlorine donor materials are depleted by the process of the present invention, heating during any subsequent refining process will not result in significant production of undesirable chlorinated compounds such as MCPDE.

[0200] Processes for refining, degumming, bleaching, deodorizing and fractionating are well known in the art.

[0201] By way of example, refining of vegetable oils, such as vegetable oils, typically involves physical or chemical refining.

[0202] In an effort to improve sustainability, vegetable oil processing lines in oil refineries have been retrofitted over the past few decades to minimize energy consumption (economisers) and reduce waste, however the steps in these two refining processes have remained essentially unchanged.

[0203] Physical refining is essentially a shortened form of chemical refining and was introduced in 1973 as the preferred method of palm oil refining. It can be a three-step sequential operation in which the input oil is pretreated with acid (degummed), washed by passing it through an adsorptive bleaching clay, and then steam distilled. This process allows for subsequent deacidification, deodorization, and destruction of carotenoids that are inherent to palm oil (i.e., crude oil is deep red in color, unlike other vegetable oils). Given the lack of a neutralization step in physical refining, the refined bleached (RB) oil produced from physical refineries has approximately the same free fatty acid (FFA) concentration as that found in the crude oil.

[0204] Neutralized bleached (NB) oil and RB palm oil from the chemical refinery are comparable to the pre-deodorized in all other respects.

[0205] The operation of the thermal decolorization unit is the main source of losses in the oil refining process, resulting in a 20-40% reduction in the oil volume after filtration. The process typically lasts for about 30-45 minutes and is typically carried out at temperatures between 95-110 °C under a vacuum of 27-33 mbar.

[0206] The thermally bleached oil may then be rerouted to a line to a degasser that aids in the removal of dissolved gases and moisture before being sent to a deodorization tower.

[0207] The bleaching process may involve heating the oil and washing the oil by passing it through an adsorbent bleaching clay.

[0208] The deodorization step may include steam distillation.

[0209] It is understood that a person skilled in the art can combine all features of the invention disclosed herein without departing from the scope of the invention disclosed.

[0210] Preferred features and embodiments of the present invention will now be described by way of non-limiting examples.

[0211] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of chemistry, biochemistry, molecular biology, microbiology, and immunology which are within the capabilities of those skilled in the art and are explained in the literature. For example, Sambrook, J., Fritsch, EFand Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, 2nd Edition, Cold Spring Harbor Laboratory Press; Ausubel, FMet al. (1995 and periodic supplements) Current Protocols in Molecular Biology, Ch. 9, 13 and 16, John Wiley & Sons; Kahn, A. (1996) DNA Isolation and Sequencing: Essential Techniques, John Wiley & Sons; Polak, JMand McGee, J.O'D. (1990) In Situ Hybridization: Principles and Practice, Oxford University Press; Gait, MJ (1984) Oligonucleotide Synthesis: A Practical Approach, IRL Press and Lilley, DMand. Dahlberg, J.E. (1992) Methods in Enzymology: DNA Structures. See Part A: Synthesis and Physical Analysis of DNA, Academic Press. Each of these general texts is incorporated herein by reference.

[0212] [Example] Analytical procedures used in the examples Sample preparation The oil samples were serially diluted before injection. 1) First, 100 μL of each sample was transferred to a vial and 900 μL of a mixture of n-hexane:acetone (1:1 v / v) was added. The sample was vortexed for 5 to 10 seconds. 2) In a second step, 50 μL of this solution was further diluted by mixing with 950 μL of acetone. The resulting solution was vortexed for 5-10 seconds. 3) 100 μL of this latter solution was mixed with 90 μL of methanol and 10 μL of an internal standard mixture containing the following stable isotope-labeled compounds dissolved in methanol at a concentration of 2 ng / μL: 1-oleoyl, 2-linoleoyl, 3-chloropropanediol- 2 H5(OL), 1-2-dipalmitoyl 3-chloropropanediol- 2 H5(PP), 1-palmitoyl 2-oleoyl 3-chloropropanediol- 2 H5(PO), 1-palmitoyl 2-linoleoyl 3-chloropropanediol- 2 H5(PL).

[0213] LC conditions Ultra-performance liquid chromatography was performed using a Waters Acquity H-class system equipped with a silica-based octadecyl phase (Waters Acquity HSS C18, 1.7 μm; 2.1×150 mm). The solvent gradients applied are summarized in Table 3.

[0214] [Table 1]

[0215] Table 3. Details of the LC gradient applied (solvent A was 1 mM ammonium formate in methanol and solvent B was 100 μM ammonium formate in isopropanol).

[0216] MS conditions Monitoring of monochloropropanediol (MCPD) esters was performed using a ThermoFisher high-resolution mass spectrometer (Orbitrap Elite Hybrid). This platform enabled highly selective mass analysis with a working mass accuracy of approximately 2 ppm. MCPD esters were analyzed in ESI positive ion mode (ESI + Under these conditions, the observed MCPD ester ion was [M+NH4] + and [M+Na] + It was an adduct.

[0217] Data Explanation Relative quantification of MCPDE was first performed by [M+NH4] + and [M+Na] + The ion chromatograms of the adducts were extracted at each m / z value in a mass window of 10 ppm, and then the resulting peak areas were integrated at the corresponding chromatographic retention times. The abbreviations of the monitored MPCDEs are as follows: PP: dipalmitoyl MCPD ester; PO: palmitoyl-oleyl MCPD ester; OO: dioleyl MCPD ester; OL: oleyl-linoleyl MCPD ester; LL: dilinoleyl MCPD ester; PL: palmitoyl-linoleyl MPCD ester.

[0218] For all experiments, the peak heights of PP, PO, OO, OL, PL, and LL MCPD were summed, multiplied by 1000, and divided by the peak heights of the four stable isotope-labeled internal standards mentioned above, which allowed for easy and quick comparison and visualization of the relative MCPDE concentrations in the investigated samples.

[0219] Sample ampoule heat treatment Thermal treatment of the crude oil samples was carried out in a Thermo Scientific Heraeus oven (series 6100) at 230 °C for 2 h under nitrogen in sealed glass ampoules. The glass ampoules were made from glass Pasteur pipettes using a Bunsen gas burner. These conditions were chosen to replicate the thermal conditions used during the deodorization of edible oils.

[0220] Examples 1 and 2 Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to pre-purification by centrifugation after reduction testing.

[0221] 6 L of crude palm oil was melted by heating to 80° C. in a water bath. The oil was homogenized by manual shaking. 1 L aliquots were transferred to 1 L polypropylene tubes (Sorvall 1000 mL) and centrifuged at 8000 g for 15 min at 40° C. in a Thermo Scientific Heraeus Cryofuge 8500i centrifuge. The sediment-free top 90% (v / v%) oil was used for further testing.

[0222] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0223] Hydroxy acid degumming Individual crude palm oil, expeller crude sunflower oil and their 1:1 mixture were degummed by heating the oil to 80°C, then phosphoric acid >85% was added in an amount of 0.02% (v / v%) of the oil. The mixture was sheared (Silverson LM-5A) at 80°C and 1500 rpm for 5 min. Water was added (2% (v / v%) of the mixture) followed by shearing (Silverson LM-5A) for 5 min while maintaining the temperature at 80°C. Centrifuge 5804R at 15000g and 40°C for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0224] The resulting 1:1 mixture of degummed crude palm oil:crude sunflower oil and 1:1 mixture of individually degummed palm oil and sunflower oil were heat treated in ampoules as described above to simulate the formation of MCPDE and analyzed by LC-MS for MCPDE content accordingly. The effect of crude sunflower oil during the degumming process is demonstrated by the observed MCPDE concentrations shown in Figure 1. The mixtures mixed before the degumming step showed a 2-fold lower MCPD concentration than the mixtures mixed after the degumming step.

[0225] Moreover, this beneficial effect was only observable if the sunflower oil was crude and not degummed. When crude palm oil was subjected to degumming in the presence of degummed sunflower oil, a two-fold higher MCPD concentration was observed compared to the mixture that contained non-degummed crude sunflower oil (see Figure 2). This suggests that indeed the gum fraction of the crude sunflower oil is responsible for the effect observed as a reduction in the MCPD content of the oil mixture.

[0226] Example 3 Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to pre-purification by centrifugation after reduction testing.

[0227] 6 L of crude palm oil was melted by heating to 80° C. in a water bath. The oil was homogenized by manual shaking. 1 L aliquots were transferred to 1 L polypropylene tubes (Sorvall 1000 mL) and centrifuged at 8000 g for 15 min at 40° C. in a Thermo Scientific Heraeus Cryofuge 8500i centrifuge. The sediment-free top 90% (v / v%) oil was used for further testing.

[0228] Commercially available 1,2-ditricosanoyl-sn-glycerol-3-phosphocholine (cat.n.850372P) was from Avanti Polar Lipids Inc. (Alabaster, Alabama, US). The saturated long fatty acyl chains each contain 23 carbon atoms.

[0229] A 10 mL aliquot of sediment-free crude palm oil was spiked with 0.4% (w / w%) 1,2-ditricosanoyl-sn-glycerol-3-phosphocholine. The mixture was heated to 90°C for 15 min and vortexed to homogenize. The mixture was then centrifuged at 15000g and 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 10% (v / v%) and the bottom 10% (v / v%) containing solids were collected for heat treatment and subsequent LC-MS analysis.

[0230] On the other hand, the MCPD concentration calculated as described above in the Data Description section shows that addition of 1,2-ditricosanoyl-sn-glycerol-3-phosphocholine, crystallization, and centrifugation reduced the relative MCPD signal from 71.7 (reference sample with 0% phosphocholine added) to 39 (with 0.4% phosphocholine added).

[0231] Furthermore, after extraction of the MCPD signal, the ratio of MCPD observed between the top 10% and bottom 10% by volume was calculated as the accumulation factor. This accumulation factor is shown in Figure 3 for centrifuged palm oil spiked with 0% phosphocholine (reference) or 0.4% phosphocholine. The results showed that the addition of 1,2-ditricosanoyl-sn-glycerol-3-phosphocholine, crystallization, and centrifugation allowed the chlorine source to be more effectively concentrated in the lower phase from the oil, which in turn allowed the upper phase to be more efficiently purified.

[0232] Example 4 Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to pre-purification by centrifugation after reduction testing.

[0233] 6 L of crude palm oil was melted by heating to 80° C. in a water bath. The oil was homogenized by manual shaking. 1 L aliquots were transferred to 1 L polypropylene tubes (Sorvall 1000 mL) and centrifuged at 8000 g for 15 min at 40° C. in a Thermo Scientific Heraeus Cryofuge 8500i centrifuge. The sediment-free top 90% (v / v%) oil was used for further testing.

[0234] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0235] Hydroxy acid degumming The oil was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% (v / v%) of the oil. The oil was then sheared (Silverson LM-5A) at 80°C and 1500 rpm for 5 min. After adding water (2% by volume of the mixture), the mixture was sheared (Silverson LM-5A) for 15 min while maintaining the temperature at 80°C. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g and 40°C for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0236] 500 mL of crude pressed sunflower was subjected to wet acid degumming. To investigate the dose-response relationship between the amount of sunflower gum extract added and the reduction effect on the final observed MCPD concentration, gum aliquots were added in various amounts to crude palm oil before wet acid degumming using the recovered gum phase.

[0237] Sunflower gum aliquots corresponding to 2 mL, 10 mL, and 30 mL of crude sunflower oil were taken. These gums were then added to 10 mL aliquots of pre-degummed crude palm oil, corresponding to gum additions of 0.2, 1, and 3 volume equivalents.

[0238] The resulting mixture was then subjected to the same wet acid degumming process as above, followed by heat treatment and LC-MS analysis.

[0239] The MCPD concentrations observed in the heat-treated oils confirm that indeed the addition of sunflower gum before the degumming process reduces the formation of MCPD, and this effect shows a dose-response relationship.

[0240] Example 5 Industrially produced crude fish oil Industrially produced crude fish oil was obtained from Sofonil SA (Manno, Switzerland).

[0241] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0242] Analysis of total MCPD by AOCS formula method The samples were submitted to SGS (SGS Germany GmbH, Hamburg, Germany) and analyzed by the AOCS Cd 29b-13 method to determine the sum of free 2-MCPD and 3-MCPD and their respective esterified (bound) forms.

[0243] The beneficial effects of expeller-pressed sunflower oil as a supplementary oil were studied in four fish oil-containing samples, as described below.

[0244] crude fish oil A 10 mL aliquot of crude fish oil was equilibrated at 60° C. for 30 min and then centrifuged at 15000 g for 15 min at 40° C. (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 10% (v / v%) and bottom 10% (v / v%) were taken for heat treatment and subsequent MCPD analysis at SGS.

[0245] Mixture of solid crude fish oil and expeller-pressed crude sunflower oil The crude fish oil was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g for 15 min at room temperature, and the resulting solid fraction was separated and used for further processing. This solid fraction of crude fish oil was mixed with pressed crude sunflower in a 1:1 ratio, heated to 80°C, and homogenized by vortexing for 10 seconds. The resulting mixture was allowed to cool to room temperature and stored at ambient temperature for one month. The mixture was then centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g for 15 min at room temperature, and the separated solid and liquid fractions of the mixture were obtained.

[0246] Liquid and solid fractions obtained from a mixture of solid crude fish oil and pressed crude sunflower oil The above resulting liquid and solid fractions obtained from the mixture of solid crude fish oil and pressed crude sunflower oil were equilibrated at 45° C. to allow complete melting of the samples. The samples were then centrifuged at 15000 g and 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 10% (v / v%) and bottom 10% (v / v%) fractions of each sample were taken for heat treatment and subsequent MCPD analysis at SGS.

[0247] The results in Figure 5 show that only a weak effect was observed when the crude fish oil was subjected to centrifugation, with 3.8 ppm of 3-MCPD detected in the upper phase and 4.9 ppm in the lower phase, which corresponds to an accumulation of about 1.3 times in the lower phase.

[0248] However, when the solid fraction of crude fish oil was mixed with expeller-extracted crude sunflower oil, very different MCPD concentrations were observed in the liquid and solid fractions, with 1.2 ppm and 4.7 ppm of 3-MCPD, respectively. This suggests an accumulation factor of 4 between the liquid and solid fractions (see the second bar in Figure 5). This result also suggests that the source of MCPD was mostly enriched in the solid phase and the liquid phase was refined.

[0249] To demonstrate that this accumulation effect was due to the centrifugation itself, both fractions were heated to 45 °C to completely melt the solid fraction and subjected to centrifugation as described above. As expected, the upper and lower phases obtained from the liquid fraction showed the same 3-MCPD concentrations, 1.1 ppm and 1.1 ppm, respectively. However, the upper and lower phases obtained from the solid fraction showed 3-MCPD concentrations of 2.8 ppm and 7.6 ppm, respectively. This corresponds to an accumulation factor of 2.7 in the lower phase (see the fourth bar in Figure 5), confirming that the centrifugation did indeed enrich the precursor of MCPD in the lower phase.

[0250] Example 6 Effect of assisted degumming on bleached palm oil. The LC-MS analytical procedures were carried out as described at the beginning of the Examples section above.

[0251] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. Aliquots of 40 mL were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 90% (v / v%) corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0252] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0253] Gum formation from sunflower oil. Crude pressed sunflower oil was degummed by first heating the oil to 80°C and then adding 0.02% (v / v%) phosphoric acid (≧85% grade). The mixture was sheared in a Silverson LM-5A mixer at 80°C and 1500 rpm for 5 min. 2% by volume of water was then added and the mixture was further sheared for 5 min while maintaining it at 80°C. The mixture was then transferred to a 40 mL Falcon tube and centrifuged at 15000g and 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) of the oil was removed and the bottom 4 mL of liquid phase containing the gum precipitated at the bottom of the tube was used as "gum" for further workup.

[0254] Hydroxydegumming of palm oil The oil was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% by volume of the oil. The oil was then sheared (Silverson LM-5A) at 80°C and 1500 rpm for 2 minutes. After adding water (2% by volume of the mixture), the mixture was sheared (Silverson LM-5A) for 2 minutes while maintaining the temperature at 80°C. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g and 40°C for 15 minutes. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0255] Bleaching of degummed palm oil The degummed palm oil was transferred to a 0.5 L round bottom rotary evaporator flask heated at 85°C in a water bath and 1% bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um. The resulting decolorized palm oil was subjected to heat treatment and LC-MS analysis as described above.

[0256] Auxiliary degumming of bleached palm oil Adding sunflower gum to palm oil The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the bleached palm oil, which in this case corresponded to adding sunflower gum obtained from 500 mL of sunflower oil to 250 mL of bleached palm oil. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 1500 rpm for 2 minutes.

[0257] Hydroxydegumming of sunflower gum and bleached palm oil blends. The mixture containing sunflower gum and bleached palm oil was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% by volume. The mixture was then sheared (Silverson LM-5A) at 80°C and 1500 rpm for 2 minutes. After addition of water (2% by volume of the mixture), the mixture was sheared (Silverson LM-5A) for 2 minutes while maintaining the temperature at 80°C. Centrifuge 5804R at 15000g and 40°C for 10 minutes, Eppendorf, VWR International GmbH, Switzerland. The top 90% (v / v%) degummed liquid phase was removed and subjected to heat treatment and LC-MS analysis as described above.

[0258] The MCPD diester concentrations observed in these bleached heat treated oils confirmed that indeed assisted degumming involving the addition of sunflower gum after the bleaching process resulted in a reduction in the formation of MCPD (Figure 6).

[0259] For Examples 7-11, ampoule heating, analytical procedures, and data interpretation of the LC-MS analyses can be found above. Where indicated, samples were sent to an external laboratory, SGS (SGS Germany GmbH, Hamburg, Germany), for confirmatory analysis by the official AOCS Cd 29b-13 method. This method determined the sum of free 2-MCPD and 3-MCPD and their respective esterified (bound) forms.

[0260] Example 7 Effect of the second supplementary degumming The key steps of this example are summarized as follows:

[0261] [Table 2]

[0262] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. 40 mL aliquots were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 80% by volume corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0263] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through a filter paper (Whatman 595 1 / 2) at room temperature.

[0264] Gum formation from sunflower oil. Crude pressed sunflower oil was degummed by first heating the oil to 80°C and then adding 0.02% (v / v%) phosphoric acid (≧85% grade). The mixture was sheared in a Silverson LM-5A mixer at 80°C and 1500 rpm for 5 min. 2% by volume of water was then added and the mixture was further sheared for 5 min while maintaining it at 80°C. The mixture was then transferred to a 40 mL Falcon tube and centrifuged at 15000g, 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 95% (v / v%) of the oil was removed and the remaining lower phase, including the gum precipitated at the bottom of the tube, was used as "gum" for further workup.

[0265] Hydroxydegumming of palm oil The oil was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% by volume of the oil. The oil was then sheared (Silverson LM-5A) at 80°C and 1500 rpm for 2 minutes. After adding water (2% by volume of the mixture), the mixture was sheared (Silverson LM-5A) for 2 minutes while maintaining the temperature at 80°C. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g and 40°C for 15 minutes. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0266] First auxiliary degumming step Addition of sunflower gum to degummed palm oil. The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the acid-water degummed palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of degummed palm oil. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 5000 rpm for 4 minutes.

[0267] Hydroxydegumming of sunflower gum and palm oil blends. The above mixture containing sunflower gum and acid-water degummed palm oil was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% by volume. The mixture was then sheared (Silverson LM-5A) at 80°C and 5000 rpm for 2 minutes. After water was added (2% by volume of the mixture), the mixture was sheared (Silverson LM-5A) for 2 minutes while maintaining the temperature at 80°C. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g and 40°C for 15 minutes. The top 90% (v / v%) degummed liquid phase (auxiliary degummed palm oil) was removed for decolorization.

[0268] Bleaching Clay Cleaning 3 g of clay (w / w) was mixed with 97 g of Milli-Q water, shaken manually, and then centrifuged at 4500 g for 10 min to remove the water, three times in succession, and the wet clay was then dried overnight in an oven at 50°C.

[0269] Bleaching of auxiliarily degummed palm oil. The assisted degummed palm oil was transferred to a 0.25L round bottom rotary evaporator flask heated at 85°C in a water bath and 2% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature was increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0270] Auxiliary degumming after bleaching of bleached palm oil Adding sunflower gum to palm oil The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the bleached palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of bleached palm oil. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 5000 rpm for 4 minutes. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 5000 rpm for 30 minutes and then heated in a Thermo Scientific Heraeus oven (series 6100) at 165° C. for 1 hour. It is noted that this heat treatment does not yet induce the formation of MCPD.

[0271] Hydroxy acid degumming and washing of sunflower gum and bleached palm oil blends. The mixture containing sunflower gum and bleached palm oil was heated to 80°C and then phosphoric acid > 85% was added in an amount of 0.02% by volume. The mixture was then sheared at 5000 rpm at 80°C for 2 minutes (Silverson LM-5A). After water was added (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while the temperature was maintained at 80°C (Silverson LM-5A). It was centrifuged at 15000g and 40°C for 15 minutes (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) degummed liquid phase was removed for further washing. The mixture was heated to 80°C and after water was added (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while the temperature was maintained at 80°C (Silverson LM-5A). Centrifuged at 15000g, 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) was removed for a final washing step. The mixture was heated to 80°C and water was added (2% by volume of the mixture) before shearing the mixture at 5000 rpm for 2 min while maintaining the temperature at 80°C (Silverson LM-5A). Centrifuged at 15000g, 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) liquid phase (bleached and auxiliarily degummed palm oil) was removed for drying, bleaching, ampoule heat treatment and LC-MS analysis.

[0272] Oil drying The oil was transferred to a 0.5 L, 0.2 L rotary evaporator round bottom flask heated in a water bath at 85° C. The flask was rotated at 240 rpm, the temperature increased and held at 95° C. and a vacuum of 15 mbar was applied for 20 minutes.

[0273] Secondary bleaching of twice sub-degummed palm oil. After bleaching, the degummed palm oil was transferred to a 0.25L round bottom rotary evaporator flask heated at 85°C in a water bath and 2% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature was increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0274] The absolute concentration of 3-MCPD was determined by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 7 show that applying a second post-bleaching auxiliary degumming with drying results in about a 50% reduction in 3-MCPD, which in this case corresponds to a reduction of about 0.7 ppm. This effect is maintained, albeit slightly lower, when the sample is subjected to a second final bleaching process, maintaining about a 30% reduction in 3-MCPD, which in this case corresponds to a reduction of about 0.5 ppm (see the third column of Figure 7).

[0275] Example 8 Effect of dual-assisted degumming combined with washing The key steps of this example are summarized as follows:

[0276] [Table 3]

[0277] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. Aliquots of 40 mL were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 80% (v / v%) corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0278] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through a filter paper (Whatman 595 1 / 2).

[0279] Gum formation from sunflower oil. Crude pressed sunflower oil was degummed by first heating the oil to 80°C and then adding 2% (v / v%) water. The mixture was sheared in a Silverson LM-5A mixer at 80°C and 1500 rpm for 10 min. The mixture was then transferred to a 40 mL Falcon tube and centrifuged at 15000g and 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 95% (v / v%) of the oil was removed and the remaining lower phase, including the gums precipitated at the bottom of the tube, was used as "gum" for further workup.

[0280] Water degumming of palm oil The oil was heated to 80° C. and water was added in an amount of 2% by volume of the oil. The oil was then sheared (Silverson LM-5A) at 80° C. and 1500 rpm for 4 min. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g and 40° C. for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0281] First auxiliary degumming step Addition of sunflower gum to degummed palm oil. The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the water-degummed palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of water-degummed palm oil. The mixture was homogenized by shearing at 80° C. and 5000 rpm for 30 minutes in a Silverson LM-5A and then heated at 165° C. for 1 hour in a Thermo Scientific Heraeus oven (series 6100). It is noted that this heat treatment does not yet induce the formation of MCPD.

[0282] Degumming and washing of gum-palm oil mixtures After this heat treatment, the samples were cooled in a 40° C. water bath for 10 min and then subjected to centrifugation at 15000 g for 15 min at 40° C. (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) liquid phase (auxiliary degummed palm oil) was removed for further processing.

[0283] The mixture was heated to 80°C and then phosphoric acid >85% was added in an amount of 0.02% by volume. The mixture was then sheared at 5000 rpm at 80°C for 2 minutes (Silverson LM-5A). After adding water (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while maintaining the temperature at 80°C (Silverson LM-5A). Centrifuged at 15000g and 40°C for 15 minutes (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) degummed liquid phase was removed for further washing. The mixture was heated to 80°C and after adding water (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while maintaining the temperature at 80°C (Silverson LM-5A). Centrifuged at 15000g, 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) was removed for the final washing step. The mixture was heated to 80°C and water was added (2% by volume of the mixture) before shearing the mixture at 5000 rpm for 2 min while maintaining the temperature at 80°C (Silverson LM-5A). Centrifuged at 15000g, 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) liquid phase (auxiliary degummed washed palm oil) was removed for further processing.

[0284] Bleaching Clay Cleaning 3 g of clay (w / w) was mixed with 97 g of Milli-Q water, shaken manually, and then centrifuged at 4500 g for 10 min to remove the water, three times in succession, and the wet clay was then dried overnight in an oven at 50°C.

[0285] Bleaching of auxiliarily degummed palm oil. The assisted degummed palm oil was transferred to a 0.25L round bottom rotary evaporator flask heated at 85°C in a water bath and 2% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature was increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0286] Auxiliary degumming after second bleaching of bleached palm oil. This step is a repeat of the previous auxiliary degumming and washing step, but after the first bleaching.

[0287] Adding sunflower gum to palm oil The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the bleached palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of bleached palm oil. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 5000 rpm for 4 minutes. The mixture was homogenized by shearing in a Silverson LM-5A at 80° C. and 5000 rpm for 30 minutes and then heated in a Thermo Scientific Heraeus oven (series 6100) at 165° C. for 1 hour. It is noted that this heat treatment does not yet induce the formation of MCPD.

[0288] Hydroxy acid degumming and washing of sunflower gum and bleached palm oil blends. The mixture containing sunflower gum and bleached palm oil was heated to 80°C and then phosphoric acid > 85% was added in an amount of 0.02% by volume. The mixture was then sheared at 5000 rpm at 80°C for 2 minutes (Silverson LM-5A). After water was added (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while the temperature was maintained at 80°C (Silverson LM-5A). It was centrifuged at 15000g and 40°C for 15 minutes (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) degummed liquid phase was removed for further washing. The mixture was heated to 80°C and after water was added (2% by volume of the mixture), the mixture was sheared at 5000 rpm for 2 minutes while the temperature was maintained at 80°C (Silverson LM-5A). Centrifuged at 15000g for 15 min at 40°C (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) was removed for a final washing step. The mixture was heated to 80°C and water was added (2% by volume of the mixture) before shearing the mixture at 5000 rpm for 2 min while maintaining the temperature at 80°C (Silverson LM-5A). Centrifuged at 15000g for 15 min at 40°C (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) liquid phase (double assisted degummed and bleached palm oil) was removed for drying, bleaching, ampoule heat treatment and LC-MS analysis.

[0289] Oil drying The oil was transferred to a 0.5 L or 0.2 L rotary evaporator round bottom flask heated in a water bath at 85° C. The flask was rotated at 240 rpm, the temperature was increased and held at 95° C. and a vacuum of 15 mbar was applied for 20 minutes.

[0290] Secondary bleaching of twice sub-degummed palm oil. After bleaching, the degummed palm oil was transferred to a 0.25L round bottom rotary evaporator flask heated at 85°C in a water bath and 2% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature was increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0291] The MCPD diester concentrations observed in these dried, bleached, heat-treated oils show a reduction of about 77% of 3-MCPD in the dried samples due to the application of two auxiliary degumming steps with a bleaching step in between (Figure 8). This effect is maintained, albeit slightly reduced, when the samples are subjected to a second, final bleaching process, maintaining a reduction of about 71% of 3-MCPD (see the third column of Figure 8).

[0292] For the same samples, the absolute concentration of 3-MCPD was determined by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 9 confirm that applying a second post-bleaching auxiliary degumming with drying results in about a 67% reduction in 3-MCPD, which in this case corresponds to a reduction of about 1.2 ppm. This effect is maintained, albeit slightly lower, when the samples are subjected to a second final bleaching process, maintaining a reduction of about 61% in 3-MCPD, which in this case corresponds to a reduction of about 1.1 ppm (see the third column of Figure 9).

[0293] Example 9 Effect of acid-free assisted degumming combined with washing The key steps of this example are summarized as follows:

[0294] [Table 4]

[0295] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. Aliquots of 40 mL were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 80% (v / v%) corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0296] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through a filter paper (Whatman 595 1 / 2).

[0297] Gum formation from sunflower oil. Crude pressed sunflower oil was degummed by first heating the oil to 80°C and then adding 2% (v / v%) water. The mixture was sheared in a Silverson LM-5A mixer at 80°C and 1500 rpm for 10 min. The mixture was then transferred to a 40 mL Falcon tube and centrifuged at 15000g and 40°C for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 95% (v / v%) of the oil was removed and the remaining lower phase, including the gums precipitated at the bottom of the tube, was used as "gum" for further workup.

[0298] Water degumming of palm oil The oil was heated to 80° C. and water was added in an amount of 2% by volume of the oil. The oil was then sheared (Silverson LM-5A) at 80° C. and 1500 rpm for 4 min. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g and 40° C. for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0299] Assisted degumming Addition of sunflower gum to degummed palm oil. The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the water-degummed palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of water-degummed palm oil. The mixture was homogenized by shearing at 80° C. and 5000 rpm for 30 minutes in a Silverson LM-5A and then heated at 165° C. for 1 hour in a Thermo Scientific Heraeus oven (series 6100). It is noted that this heat treatment does not yet induce the formation of MCPD.

[0300] Degumming and washing of gum-palm oil mixtures After this heat treatment, the samples were cooled in a 40° C. water bath for 10 min and then subjected to centrifugation at 15000 g for 15 min at 40° C. (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) liquid phase (auxiliary degummed palm oil) was removed for further processing.

[0301] The mixture was heated to 80° C. and water was added (2% by volume of the mixture) before shearing (Silverson LM-5A) at 5000 rpm for 2 min while maintaining the temperature at 80° C. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g and 40° C. for 15 min. The top 90% (v / v%) was removed for further workup and this washing process was repeated two more times.

[0302] Oil drying The oil was transferred to a 0.5 L or 0.2 L rotary evaporator round bottom flask heated in a water bath at 85° C. The flask was rotated at 240 rpm, the temperature was increased and held at 95° C. and a vacuum of 15 mbar was applied for 20 minutes.

[0303] Bleaching Clay Cleaning 3 g of clay (w / w) was mixed with 97 g of Milli-Q water, shaken manually, and then centrifuged at 4500 g for 10 min to remove the water, three times in succession, and the wet clay was then dried overnight in an oven at 50°C.

[0304] Bleaching of degummed palm oil The assisted degummed palm oil was transferred to a 0.25L round bottom rotary evaporator flask heated at 85°C in a water bath and 2% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm, the temperature was increased and held at 95°C and a vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0305] The absolute concentration of 3-MCPD was determined by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 10 confirm that the application of acid-free assisted degumming together with drying results in about a 42% reduction in 3-MCPD, which in this case corresponds to a reduction of about 0.7 ppm. This effect is maintained when the sample is subjected to a decolorization process, maintaining a reduction of about 47% in 3-MCPD, which in this case corresponds to a reduction of about 0.8 ppm (see the third column of Figure 10).

[0306] Example 10 Assisted Sedimentation in Crude Fish Oil Industrially produced crude fish oil Industrially produced crude fish oil was obtained from Sofonil SA (Manno, Switzerland).

[0307] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0308] Formation of precipitated gums from sunflower oil. The pressed crude sunflower oil was allowed to stand at room temperature for 4 weeks to develop a sediment layer at the bottom of the storage bottle. The top 90% of the oil was removed and the bottom 10% rich in sediment was used for further work as "precipitated sunflower gum". Immediately prior to use, the precipitated sunflower gum was heated to 165°C for 30 minutes in a Thermo Scientific Heraeus oven (series 6100) and then cooled in a 40°C water bath for 15 minutes.

[0309] Assisted sedimentation of crude fish oil with precipitated sunflower gum. A 9 mL aliquot of crude fish oil was mixed with 1 mL of heat-treated precipitated sunflower gum. The mixture was heated to 80°C in a water bath for 15 minutes and shaken manually. The mixture was then stored at room temperature for 3 weeks.

[0310] After this storage, the mixture was equilibrated in a water bath at 45° C. for 15 min and then centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g for 15 min at 40° C. The top 10% (v / v%), middle 80% and bottom 10% (v / v%) were taken for heat treatment and subsequent MCPD analysis.

[0311] The absolute concentration of 3-MCPD was determined by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 11 indicate that the bottom 10% phase has more than twice as much 3-MCPD than the rest of the oil, confirming that the MCPD precursor accumulates in the bottom phase. This is complemented by the fact that the top 10% and middle 80% of the oil show approximately 1 ppm lower 3-MCPD concentrations compared to the original feedstock.

[0312] Example 11 Accelerated assisted settling of crude fish oil Industrially produced crude fish oil Industrially produced crude fish oil was obtained from Sofonil SA (Manno, Switzerland).

[0313] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through filter paper (Whatman 595 1 / 2) in an oven at 65°C.

[0314] Formation of precipitated gums from sunflower oil. The pressed crude sunflower oil was allowed to stand at room temperature for 4 weeks to develop a sediment layer at the bottom of the storage bottle. The top 90% of the oil was removed and the bottom 10% rich in sediment was used for further work as "precipitated sunflower gum". Immediately prior to use, the precipitated sunflower gum was heated to 165°C for 30 minutes in a Thermo Scientific Heraeus oven (series 6100) and then cooled in a 40°C water bath for 15 minutes.

[0315] Accelerated assisted settling of crude fish oil by precipitated sunflower gum A 9 mL aliquot of crude fish oil was mixed with 1 mL of heat-treated precipitated sunflower gum. The mixture was heated to 80°C in a water bath for 15 minutes and shaken manually. The mixture was then stored at a temperature of 6°C for one week.

[0316] After this storage, the mixture was equilibrated in a water bath at 45° C. for 15 min and then centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g for 15 min at 40° C. The top 10% (v / v%), middle 80% and bottom 10% (v / v%) were taken for heat treatment and subsequent MCPD analysis.

[0317] The absolute concentration of 3-MCPD was determined by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 12 indicate that approximately twice as much 3-MCPD occurred in the bottom 10% phase as in the rest of the oil, confirming that the MCPD precursor was accumulated in the bottom phase.

[0318] A description of the ampoule heating and analysis procedure can be found above.

[0319] Example 12 Effect of two-step assisted degumming on dried oils. The key steps of this example are summarized in the following schematic diagram.

[0320] [Table 5]

[0321] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. Aliquots of 40 mL were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 90% by volume corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0322] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through a filter paper (Whatman 595 1 / 2) at room temperature.

[0323] Gum formation from sunflower oil. Crude pressed sunflower oil was first degummed by adding 2% by volume of water at room temperature. The mixture was sheared in a Silverson LM-5A mixer at 1500 rpm for 10 min at room temperature. The mixture was then transferred to a 40 mL Falcon tube, left to stand at room temperature for 30 min, and then centrifuged at 15000 g for 15 min at room temperature (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) of the oil was removed and the remaining lower phase, including the gum precipitated at the bottom of the tube, was used as "gum" for further workup.

[0324] Water degumming of palm oil The oil was heated to 80° C. and then 2% by volume of water heated to 80° C. was added. The oil was then sheared (Silverson LM-5A) at 80° C. and 1500 rpm for 4 min. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g and 40° C. for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0325] Auxiliary degumming with one water wash step The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the water-degummed palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of degummed palm oil. The mixture was homogenized by shearing at 80° C. and 5000 rpm for 30 minutes in a Silverson LM-5A, then heated in a Thermo Scientific Heraeus oven (series 6100) at 165° C. for 1 hour with manual shaking for 10 seconds at 10 minute intervals in a closed glass container. It is noted that this heat treatment does not yet induce the formation of MCPD.

[0326] After heating, the mixture was kept at room temperature for 5 min and then cooled by placing in a room temperature water bath for 10 min. The mixture was then equilibrated at 40° C. for 10 min and centrifuged at 15000 g for 15 min at 40° C. (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) degummed liquid phase was removed and used for further workup.

[0327] In the next step, 2% (v / v%) water was added to the top 90% of the degummed liquid phase. The mixture was then sheared (Silverson LM-5A) at 5000 rpm for 4 min at 80°C and equilibrated at 40°C for 10 min. It was then centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g for 15 min at 40°C. The top 90% (v / v%) degummed / washed liquid phase (auxiliary degummed palm oil) was removed for further processing.

[0328] Oil drying The oil was transferred to a 0.2 L round bottom rotary evaporator flask heated in a water bath at 95° C. The flask was rotated at 240 rpm and vacuum was applied at 20 mbar for 20 minutes.

[0329] The samples obtained according to the above schematic were subjected to analysis by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 13 confirm that applying assisted degumming without any bleaching process results in about a 55% reduction of 3-MCPD in the dried oil, which in this case corresponds to a reduction of about 1.7 ppm. An even greater effect was observed when the assisted degumming was repeated, reaching a final concentration of 0.93 ppm of 3-MCPD (see column "Double (assisted degummed / dried) oil" in Figure 13).

[0330] Example 13 Effect of single and double assisted degumming in combination with bleaching The key steps of this example are summarized in the following schematic diagram.

[0331] [Table 6]

[0332] Industrially produced crude palm oil Industrially produced crude palm oil was purchased from Nutriswiss (Lyss, Switzerland). The oil was subjected to preliminary purification by centrifugation. 5 L of industrial crude palm oil was equilibrated at 60° C. in a water bath for 30 min and then vigorously homogenized by manual shaking. Aliquots of 40 mL were transferred to 50 mL Falcon® tubes and centrifuged at 15000 g, 40° C. for 15 min (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The sediment-free top 90% by volume corresponding to 36 mL aliquots was taken from each Falcon® tube and combined for further workup.

[0333] Expeller pressed sunflower oil Sunflower seeds were pressed using a domestic electric oil press (OP700, Rommelsbacher, Germany) to obtain pressed oil and the remaining solid residue (cake). The pressed oil was then filtered through a filter paper (Whatman 595 1 / 2) at room temperature.

[0334] Gum formation from sunflower oil. Crude pressed sunflower oil was first degummed by adding 2% by volume of water at room temperature. The mixture was sheared in a Silverson LM-5A mixer at 1500 rpm for 10 min at room temperature. The mixture was then transferred to a 40 mL Falcon tube, left to stand at room temperature for 30 min, and then centrifuged at 15000 g for 15 min at room temperature (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) of the oil was removed and the remaining lower phase, including the gum precipitated at the bottom of the tube, was used as "gum" for further workup.

[0335] Water degumming of palm oil The oil was heated to 80° C. and then 2% by volume of water heated to 80° C. was added. The oil was then sheared (Silverson LM-5A) at 80° C. and 1500 rpm for 4 min. It was centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000 g and 40° C. for 15 min. The top 90% (v / v%) degummed liquid phase was used for further workup.

[0336] Auxiliary degumming with one water wash step The gum was obtained from sunflower oil as described above. Two volume equivalents of sunflower gum were then added to the water-degummed palm oil. In this case, this addition corresponded to adding sunflower gum obtained from 80 mL of sunflower oil to 40 mL of degummed palm oil. The mixture was homogenized by shearing at 80° C. and 5000 rpm for 30 minutes in a Silverson LM-5A, then heated in a Thermo Scientific Heraeus oven (series 6100) at 165° C. for 1 hour with manual shaking for 10 seconds at 10 minute intervals in a closed glass container. It is noted that this heat treatment does not yet induce the formation of MCPD.

[0337] After heating, the mixture was kept at room temperature for 5 min and then cooled by placing in a room temperature water bath for 10 min. The mixture was then equilibrated at 40° C. for 10 min and centrifuged at 15000 g for 15 min at 40° C. (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland). The top 90% (v / v%) degummed liquid phase was removed and used for further workup.

[0338] In the next step, 2% (v / v%) water was added to the top 90% of the degummed liquid phase. The mixture was then sheared (Silverson LM-5A) at 5000 rpm for 4 min at 80°C and equilibrated at 40°C for 10 min. It was then centrifuged (Centrifuge 5804R, Eppendorf, VWR International GmbH, Switzerland) at 15000g for 15 min at 40°C. The top 90% (v / v%) degummed / washed liquid phase (auxiliary degummed palm oil) was removed for further processing.

[0339] Oil drying The oil was transferred to a 0.2 L round bottom rotary evaporator flask heated in a water bath at 95° C. The flask was rotated at 240 rpm and vacuum was applied at 20 mbar for 20 minutes.

[0340] Bleaching Clay Cleaning 3 g of clay (w / w) was mixed with 97 g of Milli-Q water, shaken manually, then centrifuged at 4500 g for 10 min and the water was removed by pipetting, three times in succession. The wet clay was then dried in an oven at 50° C. for 24 h.

[0341] Bleaching of auxiliarily degummed palm oil. The assisted degummed palm oil was transferred to a 0.2 L round bottom rotary evaporator flask heated at 95°C in a water bath and 2 wt% pre-washed and dried bleaching earth (Tonsil 112FF) was added. The flask was rotated at 240 rpm and vacuum was applied at 50 mbar for 20 minutes. Finally, the oil was filtered through a vacuum Millipore filtration apparatus using a Whatman filter 8um.

[0342] The samples obtained according to the above schematic were subjected to analysis by the AOCS official Cd29b-13 method in the SGS laboratory. The results shown in Figure 14 confirm that applying auxiliary degumming in combination with the bleaching process results in about a 60% reduction in 3-MCPD, which in this case corresponds to a reduction of about 1.4 ppm. The greatest effect was observed when auxiliary degumming and bleaching were repeated in succession, reaching a final concentration of 0.64 ppm of 3-MCPD (see column E of Figure 14 "Double (auxiliary degummed / bleached) oil"). For the latter, this corresponds to a reduction of about 73% (see columns A and E).

[0343] All publications mentioned in the above specification are incorporated herein by reference. Various modifications and variations of the methods, uses and products disclosed in the present invention will be apparent to those skilled in the art without departing from the scope and spirit of the present invention. Although the present invention has been disclosed in connection with certain preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the disclosed modes for carrying out the invention, which will be apparent to those skilled in the art, are intended to be within the scope of the following claims.

Claims

1. 1. A method for preventing or reducing the formation of monochloropropanediol (MCPD) or monochloropropanediol esters (MCPDE) in a triacylglyceride oil, comprising: (a) the triacylglyceride oil, 1. A supplementary oil having a higher phospholipid content and / or wax content than the triacylglyceride oil; and / or 2. Mixing with gum extract from oil; (b) degumming the triacylglyceride oil blend to form an insoluble component, and optionally crystallizing the insoluble component; (c) removing the insoluble or crystallizable component by 1. by subjecting the triacylglyceride oil mixture to centrifugal force; and / or 2. By allowing the insoluble or crystallized components to settle by gravity, concentrating said triacylglyceride oil blend; (d) separating said insoluble or crystallized components from said triacylglyceride oil blend and, optionally, subjecting said insoluble or crystallized components to one or more processes selected from degumming, physical refining, chemical refining, neutralization, interesterification, bleaching, winterization, and fractionation; (e) applying a heat treatment to the triacylglyceride oil blend, the triacylglyceride oil is a vegetable oil, an animal oil, a fish oil, a yeast oil, a fungal oil or an algae oil; the adjunct oil is a crude oil, such as sunflower oil, corn oil, canola oil, soybean oil, or high oleic variants thereof; The method, wherein said gum extract is obtained from crude oil, such as sunflower oil, corn oil, canola oil, soybean oil, rapeseed oil, or high oleic variants thereof.

2. 2. The method of claim 1, wherein the triacylglyceride oil is palm oil or a fraction obtained from palm oil.

3. The method of claim 1 , wherein the triacylglyceride oil is a fish oil or a fraction obtained from a fish oil.

4. 4. The method of any one of claims 1 to 3, wherein the insoluble or crystallized components are separated from the triacylglyceride oil blend by one or more of filtration, decantation, centrifugation, pumping, and draining.

5. 5. The method according to any one of claims 1 to 4, wherein the gum extract is obtained by one or more of precipitation, crystallization and sedimentation, and centrifugation.

6. 6. A method according to any one of claims 1 to 5, wherein the gum extract is obtained by one or more of water degumming and acid degumming.

7. 3. The method of claim 2, wherein the triacylglyceride oil is crude palm oil and the gum extract of step (a2) is a gum extract obtained from sunflower oil.

8. The method described in any one of claims 1 to 7, wherein the triacylglyceride oil is a crude oil.

9. 7. The method according to any one of claims 1 to 6, wherein the triacylglyceride oil in step (a) is a triacylglyceride oil mixed with water and / or an acid, a base, or a salt.

10. 7. The method according to any one of claims 1 to 6, wherein the triacylglyceride oil is degummed and / or neutralized and / or bleached prior to mixing.

11. 11. The method according to any one of claims 1 to 10, wherein the triacylglyceride oil has a crystallized triacylglycerol content of less than 10% (w / w%) prior to blending.

12. The method according to any one of claims 1 to 11, wherein the triacylglyceride oil is free of added ionic, cationic and anionic surfactants and / or additives prior to mixing.

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