Process for the preparation of human milk fat substitutes

Abstract

Disclosed are human milk fat (HMF) substitutes, processes for preparation thereof, uses thereof, and fat blends and formula containing the HMF substitutes. A fat base composition of the invention includes a mixture of vegetable-derived triglycerides, with less than 50% of the fatty acid residues bonded at the sn-2 position being saturated; and/or with the amount of saturated fatty acid residues bonded at the sn-2 position of the glycerol backbone less than about 43.5% of the total amount of the fatty acid residues. Typically, substantially all of the saturated fatty acids bonded at the sn-2 position of the glycerol backbone are palmitic acid residues. Also disclosed are substitute HMF compositions including a blend of at least 25% or at least 30% of the fat base composition of the invention with up to 75%, or respectively up to 70%, of at least one vegetable oil. Processes for preparing the fat base compositions and blend are also disclosed. Further disclosed are infant formulas including the fat base composition or substitute human milk fat composition.

Claims

1. A process for the preparation of a substitute human milk fat (HMF) composition comprising admixing (i) a fat base composition that is a mixture of enzymatically structured triglycerides, with (ii) at least one vegetable oil, wherein said (i) fat base composition and said (ii) at least one vegetable oil are mixed at a ratio of (i):(ii) of from 0.13:1 to 4:1, wherein the vegetable oil is enzymatically randomized prior to being mixed with the fat base composition; and wherein said fat base composition is a mixture of structured triglycerides, comprising 0-20% C12:0 fatty acids of the total fatty acids; 0-15% C14:0 fatty acids of the total fatty acids; 20-55% C16:0 fatty acids of the total fatty acids, of which above 38% are esterified at the sn-2 position of the glycerol backbone of said triglycerides; 1-7% C18:0 fatty acids of the total fatty acids; 25-65% C18:1 fatty acids of the total fatty acids; 2-40% C18:2 fatty acids of the total fatty acids; 0-8% C18:3 fatty acids of the total fatty acids; other fatty acids are present in levels of less than 8% of the total fatty acids.

2. The process of claim 1, wherein said fat base has 10-15% C12:0 fatty acids out of the total fatty acids; 5-10% C14:0 fatty acids out of the total fatty acids; 3-5% C18:0 fatty acids out of the total fatty acids; 30-40% C18:1 fatty acids out of the total fatty acids; 10-20% C18:2 fatty acids out of the total fatty acids; and 1.5-3% C18:3 fatty acids out of the total fatty acids.

3. The process of claim 1, wherein said at least one vegetable oil is enzymatically randomized to have 33% of the palmitic acid residues thereof esterified at the sn-2 position of the glycerol backbone of the triglycerides of said at least one vegetable oil.

4. The process of claim 1, wherein said vegetable oil is selected from the group consisting of soy oil, palm tree oil, canola oil, coconut oil, palm kernel oil, sunflower oil, corn oil and rapeseed oil.

5. An infant formula comprising the substitute human milk fat composition prepared by the process of claim 1 and at least one protein component.

6. The infant formula of claim 5, further optionally comprising at least one of vitamins, minerals, nucleotides, amino acids and carbohydrates.

Description

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

(1) In search for nutritionally beneficiary, yet cheap to produce HMF substitutes, the present inventors have developed novel fat-base compositions, that are characterized by having less than 50% of their sn-2 fatty acids as saturated. These novel HMF substitutes can be produced by interesterification of relatively low-palmitic acid triglycerides. The fat-base compositions can be used for preparing fat blends (final, ready for use as infants formulas components) which are characterized by a total saturated (specifically palmitic acid) fatty acids level similar to HMF and a ratio of sn-2 saturated (substantially palmitic) fatty acids to total saturated (palmitic) fatty acids higher than 33% and preferably lower than 43.5%, more preferably lower than 40%. This fat-base composition, the process for its preparation and its various uses constitute the first aspect of the present invention.

(2) Thus, in a first embodiment, the present invention relates to the production of a fat concentrate by interesterification process, in which the starting material is not as rich in palmitic acid, as described in the prior art, but a combination of vegetable oils as the triglyceride raw material, having a palmitic acid content of about 50% and total saturated fatty acids content lower than 60%. The compositions of two exemplary triglyceride starting materials are shown in Table 1.

(3) TABLE-US-00001 TABLE 1 Triglycerides Palm Palm % % stearin stearin Palm C16 saturated % w/w % w/w oil Rapeseed from from in blend in blend % w/w Oil % total total Iodine Iodine in w/w fatty fatty Value 34 Value 15 blend in blend acids acids Example 1 20 80 51 57.3 Example 2 78 22 48.7 53.7

(4) These raw materials can be used as starting material in an interesterification process, employing suitable specific 1,3-lipases, to produce a fat-base composition, which in turn can be used to produce HMF substitutes (replacers), such as those described above. These substitutes provide limited similarity to HMF by having a ratio of sn-2 palmitic acid to total palmitic acid higher than 33%, thus improving the infant's absorption of calcium and of fatty acids, used as an energy source. These HMF substitutes of the present invention exhibit improved calcium absorption over simple blends of vegetable oils used in infant formulas, have a maximal sn-2 palmitic acid to total palmitic acid ratio of preferably 1:3 (33%), and are produced at lower cost compared to HMF substitutes produced from prior art fat-bases, such as those having a ratio of sn-2 palmitic acid to total palmitic of over 50%.

(5) The low-palmitic raw material triglycerides described above may be randomized before the interesterification (as described below) and further reacted with a free fatty acids mixture containing relatively high quantities of oleic acid, at ratios of 1.5:1 (FFA to triglycerides) as in Example 1, or even as low as 1:1, as in Example 2. It may be noted that prior art compositions usually have ratios of 5:1 or as low as 3:1 (FFA to fat). The fatty acid composition (FAC) of the fat bases produced from the triglycerides of Examples 1 and 2 is described in Table 2.

(6) TABLE-US-00002 TABLE 2 Example 2 % at Example 1 sn-2 % in % at sn-2 out of sn-2 % out out of % out total from of total total sn- % in sn-2 of total sn-2 total Fatty fatty 2 fatty from total fatty fatty fatty acid acids acids fatty acid acids acids acid C12 0.8 0.4 16.7 0.2 0.2 33.3 C14 0.7 0.9 42.9 0.6 0.9 50.0 C16 28.4 40.8 47.9 28.1 40.4 47.9 C18 3.6 3.9 36.1 3.6 3.8 35.2 C18:1 51.3 42.1 27.4 54.5 42.2 25.8 C18:2 12.3 10.4 28.2 10.8 10 30.9 C18:3 1.6 0.6 12.5 1.1 1.5 45.5 C20:0 0.3 0.3 33.3 0.3 0.3 33.3 C20:1 0.2 0.5 0.6 Total 33.5 46.3 46.1 32.5 45.6 46.8 SAFA Abbreviations: FAC, fatty acid composition; SAFA, Saturated Fatty Acids; FFA, free fatty acid.

(7) In both examples the resulting fat-base has a sn-2 saturated fatty acid (SAFA) content of less than 50% and the ratio of sn-2 palmitic to total palmitic is also below 50%.

(8) These fat bases can be further blended with vegetable oils to produce a final HMF substitute/replacer, with a sn-2 saturated FA to total saturated FA ratio higher than 33% (39.3% when using fat base of Example 2) and sn-2 palmitic to total palmitic ratio of about 44% (44.2% when using fat base of Example 2).

(9) Tables 3 and 4 describe the blending of the fat base of Example 2, with the vegetable oils detailed therein and composition of the blend. The total palmitic acid content is 22%, comparable to the level found in HMF.

(10) TABLE-US-00003 TABLE 3 Blending oils Component % w/w in blend Fat base Example 2 58.8 Coconut oil 21.2 Palm Oil 5.9 Corn oil 8.2 Rapeseed oil 5.9

(11) TABLE-US-00004 TABLE 4 Fatty acid % out of total fatty acids C12 10.2 C14 4.3 C16 22.1 C18 3.2 C18:1 41.9 C18:2 13.1 C18:3 1.3 % C16 in 44.2 sn-2 from total C16 total SAFA 39.8 total SAFA 47.0 at sn-2 % SAFA in 39.3 sn-2 from total SAFA

(12) Thus, in accordance with this aspect of the invention there are provided a process and triglyceride compositions employed therein, wherein the triglycerides are blends of palm stearin interesterified with palm oil, palm stearin interesterified with rapeseed oil, and the like. The typical fatty acid composition of the resulting triglycerides starting material is less than 60%, preferably between about 48 to about 57% saturated fatty acids out of total fatty acids. At this stage, the maximal ratio between SAFA at the sn-2 position to total SAFA is about 1:3 (33%). The free fatty acids (FFA) used are a source rich in oleic acid and low in saturated fatty acids. Typical values are 2-5% C16:0, 60-80% C18:1, 1-4% C18:0. Possible sources are rapeseed fatty acids, palm kernel oil C18 fraction fatty acids, high oleic sunflower oil FFA and others. Also a combination between at least two different sources of fatty acids is possible. The ratio between triglycerides (TAG) and FFA is typically from about 40:60 (TAG:FFA) to about 50:50.

(13) The fatty acid used for enriching sn, positions 1 and 3 with unsaturated fatty acids can come from free fatty acids, as exemplified here, but also from alkyl esters of fatty acids, preferably methyl or ethyl esters, as well as glyceride esters.

(14) In a further aspect the current invention relates to a fat composition that is produced by interesterification of a triglycerides source with a relatively low amount of free fatty acids. In addition, the free fatty acid source is rich in both oleic acid and lauric acid. It is known from the prior art that fat bases can be produced by using a large excess of unsaturated fatty acids during the interesterification. Excesses of up to 5 parts FFA to fat were used. In most cases excesses of 3:1 or 2:1 FFA to fat, have been used. In all cases these fatty acids mixture were rich in oleic acid, as well as other C18 unsaturated fatty acids. These fatty acids mixtures did not contain saturated fatty acids, especially lauric acid. The present invention utilizes a ratio of 1.5:1 and even as low as 1:1 ratios of FFA to fat, in a continuous-bed, batch system. Furthermore, the use of fatty acids mixture which contains lauric acid enables maintaining relatively low sn-2 saturated fatty acids to total saturated fatty acids ratio, without increasing the risk of formation of insoluble calcium saturated fatty acids complexes. This can be attributed to the fact that lauric acid, although saturated, does not form insoluble calcium complexes as do the longer, C16 and up, saturated fatty acids [Tantibhedhyangkul and Hashim (1978) Pediatrics. 61(4):537-45; Finley and Davidson (1980) Pediatrics. 65(1):132-8; Lien (1994) J. Pediatr. 125(5):S62-8; Lien et al. (1997) J Pediatr Gastroenterol Nutr. 25(2): 167-74].

(15) Thus, in a second aspect the invention relates to a process for preparing fat-base concentrates characterized in using small or equi-weight amounts of oleic acid rich fatty acids mixtures, which are also relatively rich in lauric acid (above 3%). The triglycerides are rich in palmitic acid, preferably over 70%, and contain very small amounts of linolenic acid.

(16) Also in this embodiment, the palmitic-rich triglycerides may be randomized by a standard chemical or enzymatic interesterification process as described below.

(17) In this aspect of the invention, the palmitic-rich triglycerides are reacted with fatty acids mixtures which have relatively high levels of lauric acid, thus reducing the sn-2 SAFA to total SAFA ratio, whilst maintaining a high sn-2 palmitic to total palmitic ratio and not adversely affecting calcium and fatty acid intake by the infant (see above), as compared with similar blends obtained from fat bases with high sn-2 palmitic to total palmitic ratios.

(18) The ratio between the fatty acids and triglycerides in the interesterification process is from 1.5:1 (Example 3, Table 5) to 2:1 (Example 4, Table 6).

(19) TABLE-US-00005 TABLE 5 Example 3. Product Triglyceride Free fatty % at % at sn-2 (40%) acid (60%) sn-2 out of Ratio % out of % out of out of total sn-2 % in sn-2 total fatty total fatty total fatty from total Fatty acid acids acids sn-2 acids fatty acid C12 0.2 20.0 8.6 0.9 3.5 C14 1.4 0.2 0.8 0.1 4.2 C16 80.0 3.2 40.8 61.0 49.8 C18 5.0 2.0 3.9 4.0 34.2 C18:1 10.9 65.0 39.6 29.0 24.4 C18:2 2.2 9.0 6.3 4.0 21.2 C18:3 0.3 Total SAFA 54.1 66.0 40.7

(20) TABLE-US-00006 TABLE 6 Example 4. Product (B)-after FFA distillation Triglyceride Free fatty % at sn-2 Ratio (33%) acid (67%) % out of out of total % in sn-2 % out of total % out of total total fatty sn-2 fatty from total Fatty acid fatty acids fatty acids acids acids fatty acid C12 0.2 15.0 7.5 0.2 0.9 C14 1.4 0.2 0.7 0.1 4.8 C16 80.0 3.4 38.0 59.0 51.8 C18 5.0 3.0 3.8 4.0 35.1 C18:1 10.9 68.0 42.5 31.0 24.3 C18:2 2.2 10.0 7.2 5.0 23.1 C18:3 0.3 Total SAFA 50.0 63.3 42.2

(21) As can be seen in Examples 3 and 4, the resulting fat bases have sn-2 SAFA to total SAFA ratio of less than 43.5, while the sn-2 palmitic to total palmitic ratio is about 50%.

(22) The triglycerides are typically low Iodine Value (IV) palm stearin with up to 80% palmitic acid. The triglycerides can be a blend of interesterified palm stearin with palm oil, palm stearin with rapeseed oil, palm oil, etc. The ratio between sn-2 SAFA to total SAFA of the triglycerides is maximum 33%.

(23) The free fatty acids are a source rich in oleic acid, low in saturated long chain fatty acids of C14 and up, and enriched with lauric acid. Typical values are 2-5% C16:0, 60-80% C18:1, 1-4% C18:0, 5-30% C12 out of the total fatty acids. Possible sources are rapeseed fatty acids, palm kernel oil C18 fraction fatty acids, lauric oils (such as palm kernel oil and coconut oil), high oleic sunflower oil free fatty acids and others. Also a combination between two or more different sources of fatty acids is possible.

(24) Table 7 below exemplifies the preparation of a blend, based on the fat-base of Example 4, with additional vegetable oils as detailed in the Table. The admixed vegetable oils of the example were randomized prior to their blending with the fat base as described below.

(25) The fatty acid composition of the resulting blend can be seen in Table 8. The sn-2 SAFA to total SAFA of the blend is 37% while the sn-2 palmitic to total palmitic is 44%. Thus this blend is comparable in sn-2 palmitic to total palmitic ratio, total palmitic content and the consequent contribution to calcium intake to similar blends obtained from fat bases of significantly higher sn-2 palmitic to total palmitic ratios.

(26) TABLE-US-00007 TABLE 7 Preparation of blend Blending oil % in blend Product from Example 4 37.7 Coconut oil 18.9 Palm oil 9.4 Corn Oil 9.4 Rapeseed Oil 24.5 Total 99.9

(27) TABLE-US-00008 TABLE 8 Final blend % at sn-2 out of % out of total fatty total sn-2 fatty Fatty acid acid acids C12 11.8 9.0 C14 3.8 3.9 C16 22.2 29.3 C18 2.9 3.0 C18:1 38.8 34.5 C18:2 14.0 13.6 C18:3 2.7 2.6 Total SAFA 40.7 45.2

(28) In a further embodiment, the present invention relates to an interesterification fat product that can be used as the fat fraction of infant formula, without prior blending with additional vegetable or other oils, and that is characterized by a total saturated (specifically palmitic acid) fatty acids level similar to HMF and a ratio of sn-2 saturated (palmitic) fatty acids to total saturated (palmitic) fatty acids higher than 33%, containing all the necessary fatty acids required in infants nutrition.

(29) In a further aspect, the invention relates to the direct production of a HMF substitute, with a fatty acid composition and sn-2 palmitic to total palmitic ratio, and sn-2 SAFA to total SAFA ratios, similar to that of the blends described above, without the necessity to first obtain a suitable fat base, to be then blended with the other ingredients. According to this aspect of the invention, the product of the interesterification can be used as a final HMF replacer for infant formulas in terms of its fatty acid composition and ratios improved over simple blends of vegetable oils, resulting in improved calcium and fatty acid intake.

(30) This can be achieved by reacting a suitable triglyceride mixture (which may be randomized prior to the reaction, as described herein) with a fatty acid mixture characterized by its high level of oleic acid and total fatty acid profile designed to substantially mimic the sn-1,3 fatty acid profile of HMF. The triglycerides can be of high palmitic acid levels or of lower palmitic acid levels (as described above), according to the target grade and level of similarity to HMF of the desired HMF substitute.

(31) The fatty acids mixture used in the interesterification can include fatty acids from C8 to C24, including saturated such as lauric, myristic, stearic, mono-di-unsaturated, such as oleic, palmitoleic, and linoleic, and polyunsaturated, such as linolenic, arachidonic, docosahexaenoic, etc.

(32) The desired fatty acid mixture is obtained by mixing vegetable oils and fats as shown in Table 9, which serves as an example for raw triglycerides source. The separated to glycerol and free fatty acids using standard procedure. The composition of the derived free fatty acids is the same as the composition of the raw triglycerides.

(33) The ratio between the fatty acids mixtures and triglycerides in the interesterification reaction can be 5:1 to 1:2, preferably 3:1 to 1:1 and the reaction can be carried out by a continuous bed process or a fixed bed process using biocatalysts, preferably immobilized, preferably of 1,3 lipase activity.

(34) This process can yield products such as described in Applicant's said WO2005/036987 and in EP 0 209 327 (different blends).

(35) The resulting HMF substitutes can be of sn-2 palmitic to total palmitic ratios of above 33%, preferably over 40%, and up to 80%.

(36) Table 10 below exemplifies a HMF substitute that was produced using free fatty acid mixture of Example 5 (Table 9) and randomized palm olein at a ratio of 1.2:1. The sn-2 SAFA to total SAFA of the HMF substitute is 34.5% while the sn-2 palmitic to total palmitic is 45.3%. Thus, this composition is comparable in sn-2 palmitic to total palmitic, total palmitic content and the consequent contribution to calcium intake to similar blends obtained from fat bases.

(37) TABLE-US-00009 TABLE 9 Final High oleic blend- Coconut Palm sunflower Corn Rapeseed example oil oil oil oil oil 5 % w/w 44.2 2.3 14.0 9.3 30.2 100 in blend % fatty acid out of total fatty acids C12 47.7 0.2 21.1 C14 18.0 1.1 8.0 C16 8.9 43.8 3.3 10.6 4.3 7.7 C18 2.7 4.4 3.1 1.7 1.6 2.4 C18:1 7.0 39.5 83.2 29.5 61.0 36.8 C18:2 1.8 10.0 8.6 56.5 19.0 13.2 C18:3 0.9 10.4 3.2

(38) TABLE-US-00010 TABLE 10 Triglyceride Fatty acids % in the reaction mixture 45 55 Product after FFA distillation Description Free fatty % at % in acids sn-2 out sn-2 fraction % out of total from % fatty acid Randomized from of total sn-2 total out of total palm olein Example 5 fatty fatty fatty fatty acids total FAC total FAC acids acids acid C12 0 21.1 10.3 4.6 14.9 C14 1.1 8.0 4.5 2.6 19.4 C16 39.6 7.7 24.0 32.6 45.3 C18:0 4.1 2.4 3.3 3.7 38.1 C18:1 43.3 36.8 40.1 41.9 34.8 C18:2 10.4 13.2 11.8 11.0 31.2 C18:3 0 3.2 1.6 0.7 14.9 Total SAFA 44.8 39.1 42.0 43.6 34.5

(39) In a further aspect, the invention relates to the randomization of the resulting fat base triglycerides starting material and to the randomization of the admixed oils prior their blending with the fat base. Both these aspects of the invention have major advantages, exemplified and discussed in more detail below. It may be mentioned that while being a known process per se, neither randomization of triglycerides starting material for the production of nutritional products, particularly for infants, babies, toddlers and children, nor randomization of blending oils used in the preparation of infant formulas, have been described. This is one of the unique developments shown in the present application.

(40) The resulting triglycerides which serve as starting material for preparation of the fat base composition, are preferably randomized by a standard interesterification process, either chemically by using acidic or basic catalyst (e.g. sodium methoxide), or enzymatically by using a non-selective lipase. As explained above, by randomization, the saturated fatty acids at the sn-2 position will reach the percentage of the saturated fatty acids in the fat blend. The elevation in sn-2 saturated fatty acids will contribute to a fat base richer in sn-2 saturated fatty acids, and thus enable using a smaller quantity of fat base in the HMF substitute blend, and thus significant saving in costs.

(41) For example, standard palm oil contains about 40% C16 out of the total fatty acids, and only 5-20% C16 out of the total C16 are esterified at the sn-2 position. Following random interesterification of the palm oil, 33% C16 out of the total C16 are bonded at the sn-2 position.

(42) Another example is palm stearin IV 15 with 79% C16 content out of the total fatty acids, of which only 27.4% are esterified at the sn-2 position. Following chemical randomization, using sodium methoxide, the product shows 79% C16 content out of total fatty acids, with 33% C16 out of the total C16 esterified at the sn-2 position.

(43) Table 11 below compares a fat base produced using randomized palm stearin with a fat base produced using non-randomized palm stearin. Randomized and non-randomized palm stearin IV15 were reacted separately with commercial oleic free fatty acids at a ratio of 1:4 of palm stearin to free fatty acids. After the reaction ended, the catalyst was separated, the free fatty acids were removed using steam distillation and the triglyceride fraction was analyzed. Table 11 demonstrates that using non-randomized palm stearin, results in a fat base with 29.8% C16 fatty acids out of the total fatty acids, of which only 63.1% are esterified at the sn-2 position. However, by using randomized palm stearin, the percentage of C16 fatty acids out of the total fatty acids and the percentage of C16 esterified at the sn-2 position out of the total C16, elevate to 32.2% and 68.1%, respectively.

(44) TABLE-US-00011 TABLE 11 Fat base produced from non-randomized Fat base produced from palm stearin randomized palm stearin % C16 out of 29.8 32.2 total fatty acids % C16 at sn-2 56.4 65.8 % C16 at sn-2 63.1 68.1 from total C16

(45) The randomization of the triglycerides starting material is beneficial also for the production of better quality fat bases other than the fat bases described in the present invention. Thus, in another embodiment, the invention relates to a process for preparing fat base compositions in which the raw triglycerides source is randomized either chemically or enzymatically.

(46) The admixed vegetable oils used for blending are preferably randomized, either chemically or enzymatically, prior to their blending with the fat base composition. The randomization process randomly distributes the fatty acids of oil or fat between the three sn-positions on the glycerol backbone. Thus, the low sn-2 palmitic to total palmitic ratio of vegetable oils is elevated to 33%. The randomization process is carried out routinely on many oils and fats in the industry, mainly in order to change their physical properties for a specific application (e.g. changing the melting point of a fat). However, as described below, in this aspect of the invention the randomization is carried out for improvement of the sn-2 saturated to total saturated ratio of the admixed oils, thereby reducing to a minimum any negative effect of the admixed oils on the sn-2 saturated to total saturated ratio of the blend.

(47) Increasing the percentage of C16 at the sn-2 position out of the total C16 of the blending oils enables using less fat base for achieving a specific C16 percentage and a specific ratio of sn-2 palmitic to total palmitic. Since one major cost obstacle in preparing fat blends is the need to use high amounts of fat base, randomization of blending oils is a simple, yet effective way for saving costs in preparing fat blends. Table 12 shows that in order to obtain a typical formula containing about 21% C16 where 42% of them are esterified to the sn-2 position, 65% of Example 2 fat base are required when the blending oils are non-randomized. Table 13 shows that in order to obtain 21% C16 where 42% of them are esterified to the sn-2 position only 47% of the same fat base are required when the blending oils are randomized. This significant difference in the required amount of fat base emphasizes the importance of using randomized blending oils for cost saving purposes.

(48) TABLE-US-00012 TABLE 12 Non- Fat Non- random- Non- base of randomized ized randomized Final Example 2 Coconut oil Corn oil Rapeseed oil blend % w/w in 65.2 21.7 7.6 5.4 100 blend % fatty acid out of total fatty acids C8 7.3 1.6 C10 5.8 1.3 C12 0.2 47.7 10.5 C14 0.7 18.0 4.4 C16 28.1 8.9 10.6 4.3 21.3 % C16 47.9 10.0 5.0 5.0 42.4 at sn-2 out of total C16 C18 3.6 2.7 1.7 1.6 3.2 C18:1 54.5 7.0 29.5 61.0 42.6 C18:2 10.8 1.8 56.5 19.0 12.8 C18:3 1.1 0.9 10.4 1.4

(49) TABLE-US-00013 TABLE 13 Ran- Ran- Ran- domized Fat base domized domized Ran- Rape- Example Coconut Palm domized seed Final 2 oil oil Corn oil oil blend % w/w in 47.7 22.7 10.2 8.0 11.4 100 blend % fatty acid out of total fatty acids C8 7.3 1.7 C10 5.8 1.3 C12 0.2 47.7 0.2 11.0 C14 0.7 18.0 1.1 4.5 C16 28.1 8.9 43.8 10.6 4.3 21.2 % C16 at 47.9 33.0 33.0 33.0 33.0 42.4 sn-2 out of total C16 C18 3.6 2.7 4.4 1.7 1.6 3.1 C18:1 54.5 7.0 39.5 29.5 61.0 40.9 C18:2 10.8 1.8 10.0 56.5 19.0 13.2 C18:3 1.1 0.9 10.4 1.8

(50) It is important to notice that the production of the blend of Table 12 utilizes less fat base of Example 2 than the fat base needed for the production of the Table 3 blend, since the Table 12 blend has a lower percentage of C16 and a lower ratio of sn-2 palmitic to total palmitic than that of Table 3.

(51) The randomization of blending oils is beneficial also for blends made from fat bases other than the fat bases described in the present invention. The fat bases described in EP 0 209 327 or EP 0 495 456 or by applicant in said WO2005/036987 may be blended with randomized vegetable oils rather than with natural vegetable oils. This aspect of the invention enables achieving the same percentage of C16 and the same ratio of sn-2 palmitic acid residue to total palmitic acid residues as described by the above-mentioned applications, while using less fat base and thus saving in the blend production costs.

(52) Table 14 shows that in order to achieve EP 0 209 327 blend 2 composition (26% C16 52.6% of which being esterified to the sn-2 position), 50% of the fat base of Sample 1 of EP 0 209 327 are needed, using the non-randomized blending oils used in said patent. As may be seen from Table 15, by using the randomized blending oils of the present invention only 43% of the fat base of Sample 1 of EP 0 209 327 are needed for achieving 26% C16 of which 52.6% are esterified to the sn-2 position. This significant difference in the required amount of fat base (14% saving) demonstrates the importance and advantage of using randomized blending oils for cost saving purposes also for blends made from fat bases other than the fat bases described in and subject of the present invention.

(53) Table 16 shows that by using randomized blending oils it is possible to improve the percentage of sn-2 palmitic acid residues out of total palmitic acid residues in blends such as described in EP 0 209 327 (Blends 1-4).

(54) TABLE-US-00014 TABLE 14 Fat base Natural Natural Sample 1 Natural Sunflower Soybean final EP 0 209 327 PK oil oil blend % w/w in 50 10 10 30 100 blend % C16 in 59.9 52.6 sn-2 out of total C16

(55) TABLE-US-00015 TABLE 15 Fat Base Randomized Randomized Sample 1 Randomized Randomized Sunflower Soybean Final EP 0 209 327 PK Palm Olein Oil Oil Blend % w/w in 43 10 7 10 30 100 blend % fatty acid out of total fatty acids C8 3.4 0.3 C10 3.2 0.3 C12 48.4 0.2 5.0 C14 15.8 1.1 2.0 C16 44.5 8.2 39.0 6.0 10.0 26.0 % C16 in 59.9 33.0 33.0 33.0 33.0 52.6 sn-2 from total C16 C18 6.0 2.2 3.0 3.4 4.0 5.0 C18:1 41.5 15.9 42.0 24.3 23.0 33.5 C18:2 4.5 2.4 11.0 65.1 54.0 23.0 C18:3 0.2 7.5 2.3

(56) TABLE-US-00016 TABLE 16 Blend Blend Blend 1 of 2 of 3 of EP 0 EP 0 EP 0 Blend 4 of 209 327 209 327 209 327 EP 0 209 327 C16 as % in reported, 57.6 52.6 62.8 62.3 sn-2 out of using natural total C16 vegetable oils using 58.1 55.5 64.4 64.2 randomized vegetable oils

(57) The randomization of the blending oils has, inter alia, another important aspect besides cost-effectiveness. Randomized blending oils may contribute to -linolenic acid (C18:3) and linoleic acid (C18:2) distribution with higher similarity to HMF, in comparison with the natural oil contribution.

(58) Rapeseed oil and soybean oil are the main sources for vegetal linolenic acid and are commonly used in the manufacture of infant formulas. Table 17 shows the differences between HMF, natural and randomized rapeseed and soybean oils in the sn-2 distribution of linoleic and linolenic acids. In rapeseed oil 45-61% of the linolenic acid residues, and 32-47% of the linoleic acid residues are esterified at the sn-2 position. The distribution of linolenic acid and linoleic acid in rapeseed oil differs largely from that of HMF. In HMF only 5-20% of the linolenic acid and 20-23% of the linoleic acid are esterified at the sn-2 position. Randomization of rapeseed oil results in 33% of the -linolenic acid and 33% of the linoleic acid bonded at the sn-2 position, thus, improving the similarity of the fat substitute to HMF.

(59) In soybean oil, the distribution of linolenic acid (27-29% of the -linolenic acid are bonded at the sn-2 position) resembles that of randomized soybean oil (33%). However, in natural soybean oil, 40-43% of the linoleic acid are esterified at the sn-2 position, while in randomized soybean oil only 33% of the linoleic acid are esterified at the sn-2 position. In HMF only 20-23% of the linoleic acid residues are bonded at the sn-2 position. Thus, the utilization of randomized soybean oil instead of natural soybean oil improves the resemblance of linoleic acid distribution to that of HMF, while maintaining a similar -linolenic acid distribution.

(60) TABLE-US-00017 TABLE 17 Natural Natural Rapeseed Randomized Soybean Randomized HMF Oil Rapeseed Oil Oil Soybean Oil % C18:2 at 20-23 32-47 33 40-43 33 sn-2 out of total C18:2 % C18:3 at 5-20 45-61 33 27-29 33 sn-2 out of total C18:3

(61) The randomization of the admixed vegetable oils may be performed by a standard interesterification process, either chemically by using acidic or basic catalyst (e.g. sodium methoxide) or enzymatically by using a non-selective lipase.

(62) All of the HMF substitutes of the different aspects of the invention can be used as the fat fractions of infant formula and as fat fraction of baby foods, toddlers' foods, children and young people foods, as well as adult nutrition.

(63) The terms fat and lipid are used herein interchangeably.

(64) Lipids, under the scope of this invention, include triglycerides and derivatives, such as mono- and di-glycerides.

(65) The terms fatty acids, fatty acyl and fatty acid residues (and all their equivalents referring to specific fatty acids) are used herein interchangeably.

(66) The term infant formula as used herein encompasses infant formulas (for newborn to 6 months old infants), follow-up formulas (for 6-12 months old babies) and growing-up formulas (for 1-3 years old children).

(67) Disclosed and described, it is to be understood that this invention is not limited to the particular examples, process steps, and materials disclosed herein as such process steps and materials may vary somewhat. It is also to be understood that the terminology used herein is used for the purpose of describing particular embodiments only and not intended to be limiting since the scope of the present invention will be limited only by the appended claims and equivalents thereof.

(68) It must be noted that, as used in this specification and the appended claims, the singular forms a, an and the include plural referents unless the content clearly dictates otherwise.

(69) Throughout this specification and the claims which follow, unless the context requires otherwise, the word comprise, and variations such as comprises and comprising, will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

(70) The Examples herein are representative of techniques employed by the inventors in carrying out aspects of the present invention. It should be appreciated that while these techniques are exemplary of preferred embodiments for the practice of the invention, those of skill in the art, in light of the present disclosure, will recognize that numerous modifications can be made without departing from the spirit and intended scope of the invention.