NZ625371B2

NZ625371B2 - Stabilized concentrated liquid human milk fortifier

Info

Publication number: NZ625371B2
Application number: NZ625371A
Authority: NZ
Inventors: Gul Konuklar; Mustafa Vurma
Original assignee: Abbott Laboratories
Priority date: 2011-12-30
Filing date: 2012-12-03
Publication date: 2016-02-02

Abstract

Provided are shelf-stable concentrated liquid human milk fortifier compositions comprising extensively hydrolyzed casein and octenyl succinic acid (OSA) modified waxy potato starch. The OSA acts as a stabilizer for the compositions preventing the hardening of fat globules and the sedimentation of minerals in the fortifier. Further provided is a method of fortifying breast milk by mixing it with the concentrated liquid human milk fortifier compositions. nerals in the fortifier. Further provided is a method of fortifying breast milk by mixing it with the concentrated liquid human milk fortifier compositions.

Description

STABILIZED CONCENTRATED LIQUID HUMAN MILK FORTIFIER CROSS REFERENCE TO RELATED APPLICATIONS The present application hereby claims the beneﬁt of the provisional patent application Serial No. 61/581,634, ﬁled December 30, 2011, the disclosure of which is hereby incorporated by reference in its entirety.

FIELD OF THE DISCLOSURE The t sure relates to a stable, concentrated liquid human milk fortiﬁer including an ed stabilizer. More particularly, the present disclosure relates to a long term stable, trated liquid human milk fortiﬁer including octenyl succinic acid (OSA) modiﬁed waxy potato starch as a stabilizer. In some embodiments, the concentrated liquid human milk fortiﬁer additionally includes ively hydrolyzed casein as at least part of the protein component.

BACKGROUND OF THE DISCLOSURE Human milk is lly recognized as an ideal feeding for most s due to its overall nutritional composition. It is well known and generally ed that human milk provides infants with unique immunologic and developmental beneﬁts as compared generally to commercially available infant formulas.

For some infants, however, especially preterm infants, human milk does not always meet the complete nutritional needs. Although these infants still generally beneﬁt from human milk, it is often desirable to supplement their human milk feedings with additional nts in the form of a human milk fortiﬁer. Initially, these preterm infants may grow more rapidly than many of their term counterparts, and accelerated growth often requires additional nutrition, which is made possible by the use of a human milk fortiﬁer in combination with human milk. As such, the use of human milk fortiﬁers in al intensive care units has grown substantially and become a standard of practice.

Most ofthe human milk fortiﬁers described in the literature and commercially ble have been formulated as reconstitutable powders rather than liquids in order to minimize the volume displacement of human milk by the fortifier. The minimization of volume displacement by the fortifier is important as the infant can only intake a certain amount of ﬂuid per feeding. The reconstitutable human milk fortiﬁer powders may, however, carry the inherent risk of microbial contamination in some cases.

Although liquid human milk fortiﬁers have been considered as potential replacements for the reconstitutable powders, liquid human milk fortiﬁers generally displace more volume than powders and have onally been confronted with at least two additional problems that can ially shorten shelf life and commercial acceptance. The ﬁrst problem is known as creaming, whereby the fat es in the liquid nutritional composition floatto thetop of the product. If these fat globules are allowed to harden, a seal is effectively formed across the top of the liquid nutritional composition’s container. Additionally, these hard, fatty ts can block or clog feeding tubes or nipples, and can give the liquid ional composition an unappealing appearance and cause nutritional insufﬁciencies.

The second problem associated with many liquid human milk fortiﬁers is sedimentation, whereby nutrients, such as minerals, come out of solution and settle to the bottom of the liquid nutritional composition’s container. “The problem of sedimentation is made more acute where the sediment hardens into a cementous type of material known as “nondispersible sediment”. The problem with persible sediment is two-fold. First, the liquid human milk er may now be subject to nutrient deﬁciency as the persible sediment is often difﬁcult to ubilize into solution upon agitation.‘ The second problem with nondispersible sediment is that it, similar tohardened creaming deposits, can plug feeding tubes or nipples.

Additionally, due to ive digestive systems and poor tolerance in many preterm infants, it is generally advantageous to utilize hydrolyzed proteins, and desirably MARKED UP MARKED UP COPY COPY [0009a] According to a first aspect of the present invention there is provided a concentrated liquid human milk fortifier, sing: from 1% to 50% by weight protein on a dry weight basis, octenyl succinic anhydride modified waxy potato starch, fat, vitamins and minerals, wherein at least a n of the protein is extensively hydrolyzed casein. [0009b] According to a second aspect of the t invention there is p rovided a concentrated liquid human milk fortifier according to the first aspect above for use in fortifying human breast milk. [0009c] According to a third aspect of the present invention there is provi ded a method of fortifying breast milk, comprising: mixing a concentrated liquid human milk fortifier with breast milk in a volume to volume ratio of from 1:3 to 1:10, the trated liquid human milk fortifier comprising from 1% to 50% by weight protein, on a dry weight (10143670_1):JJC basis, and octenyl succinic ide modified waxy potato starch, wherein at least a n of the protein is extensively hydrolyzed casein. [0009d] According to a fourth aspect of the present invention there is provided use of a concentrated liquid human milk fortifier according to the first aspect above for fortifying breast milk. (10143670_1):JJC The disclosed liquid human milk fortiﬁers advantageously use a single stabilizer system to provide long term stable emulsions that have a very low level of sedimentation even in the presence of a high level Of insoluble calcium salts, while allowing the use of an extensively hydrolyzed protein. The liquid human milk fortiﬁer is emulsion stable without substantial mineral fallout and allows for homogeneous and precise ry of micronutrients and other minor nutrients to the human milk or infant formula. Additionally, because the disclosed liquid human milk fortiﬁer is a highly concentrated sterile liquid, the risk of microbial ination is reduced and the volume displacement in human breast milk or infant formula is minimized.

An embodiment of the disclosed concentrated liquid human milkfortiﬁer is preferably carrageenan-free, comprises from about 1% to about 50% by weight protein, on a dry weight basis, and octenyl succinic anhydride modiﬁed waxy potato starch and optionally further comprises fat, vitamins and minerals, wherein at least a portion of the protein is extensively hydrolyzed casein, preferably n the n is extensively hydrolyzed casein at 100% by weight of the total n component, more preferably wherein the extensively hydrolyzed casein at 100% by weight of the total protein component is hypoallergenic protein.

In another embodiment of the sed concentrated liquid human milk fortiﬁer, ‘ the fortiﬁer further comprises the octenyl succinic anhydride modiﬁed waxy potato starch , which is present in the concentrated liquid human milk fortiﬁer at from 0.1% to 3.5%, preferably from 0.8% to 1.5%, by weight of the er. In r embodiment, the octenyl succinic 1' anhydride d waxy potato starch is a sole stabilizer of the concentrated liquid human milk. fortiﬁer.

In another embodiment of the disclosed concentrated liquid human milk fortiﬁer, the fortiﬁer further comprises a solids content of at least 5% by weight, ably from 20% to 55% by weight; In another embodiment of the disclosed concentrated liquid human milk-fortiﬁer, the fortiﬁer is an aseptically-sterilized trated liquid human milk fortiﬁer.

In another embodiment of the disclosed concentrated liquid human milk fortiﬁer, the fortiﬁer is carrageenan-free and comprises from 1% to 50% by weight of extensively hydrolyzed hypoallergenic casein, from 0.8% to 1.5% by weight, octenyl succinic anhydride modiﬁed waxy potato starch, and a solids content of from 20% to 55% by weight. In another embodiment, the octenyl succinic anhydride modiﬁed waxy potato starch is a sole stabilizer.

'In another ment, the disclosure is directed to the use of a concentrated liquid human milk fortiﬁer according to any one of the preceding embodiments.

An embodiment ofthe present disclosure is a method of fortifying breast milk sing mixing a concentrated liquid human milk er with breast milk in a volume to volume ratio of from 1:3 to 1:10, preferably from 123.5 to 1:7, the liquid human milk fortiﬁer comprising from 1% to 50% by weight protein, on a dry weight basis, and octenyl succinic anhydride modiﬁed waxy potato starch, wherein at least a portion of the protein is extensively hydrolyzed casein, ably wherein the protein is extensively hydrolyzed casein at 100% by weight of the total protein component, more preferably wherein the extensively hydrolyzed casein at 100% by weight of the total protein component is hypoallergenic protein.

In another embodiment of the method, the er further comprises the octenyl ic anhydride d waxy potato starch which is present in the concentrated liquid human milk fortiﬁer at from 0.1% to 3.5%, preferably from 0.8% to 1.5%, by weight of the fortiﬁcr. In another embodiment, the octenyl succinic anhydride modiﬁed waxy potato starch is a sole izer of the concentrated liquid human milk fortiﬁer.

An embodimentofthe present disclosure is a method of reducing creaming and sedimentation in a nutritional , the method comprising preparing a nutritional liquid with l succinic anhydride modiﬁed waxy potato starch, wherein the nutritional liquid comprises fat, ydrate, and protein, and wherein at least a portion of the protein is extensively hydrolyzed casein.

It has been unexpectedly found that stable concentrated liquid human milk fortiﬁers and concentrated liquid ional compositions can advantageously be prepared with OSA modiﬁed waxy potato starch as a stabilizer, and in some embodiments, as the sole stabilizer. The use of OSA modiﬁed waxy potato starch provides for ed mineral suspension in combination with improved emulsion stability in the liquid human milk fortiﬁer due to its unique phySiochemical characteristics. surprisingly, when the OSA modiﬁed waxy potato starch is introduced into the liquid human milk fortiﬁer, the concentrated liquid can be ed with high levels of insoluble calcium salts and extensively hydrolyzed protein without mineral fallout or separation.

Additionally, the concentrated liquid human milk fortiﬁers and liquid nutritional compositions can be prepared to e lergenic extensively hydrolyzed casein proteins, which is beneﬁcial for use in low birth weight and preterm infants having sensitive digestive s. These liquids including extensively hydrolyzed casein proteins can be prepared without disrupting the long term stability or emulsion properties of the liquid. By preparing a concentrated liquid utilizing a OSA modiﬁed waxy potato starch as a stabilizer, the concentrated liquid may e up to 100% extensively hydrolyzed casein (by weight of the protein ent) while maintaining the desired emulsion and stability properties.

BRIEF DESCRIPTION OF THE GS depicts a sample of concentrated liquid human milk fortiﬁer including no stabilizer. depicts a sample of concentrated liquid human milk fortiﬁer including OSA modiﬁed waxy corn starch as a stabilizer. depicts a sample of concentrated liquid human milk er including OSA modiﬁed waxy potato starch as a stabilizer. [0026‘] FIGS. 4A—4C depicts samples of concentrated liquid human milk ers including various stabilizers as analyzed in Example 5. is a graph depicting elasticity behavior of various trated liquid human milk fortiﬁers as a function of strain as analyzed in e 6. is a graph depicting dynamic modula of s concentrated liquid human milk fortiﬁers as a function of frequency and temperature as analyzed in Example 7.

DETAILED DESCRIPTION OF THE DISCLOSURE The concentrated liquid human milk fortiﬁers of the present disclosure generally have a high solids content and extensively hydrolyzed casein in combination with OSA d waxy potato starch. The liquid human milk fortiﬁers of the present disclosure address and e a solution to the longstanding problem of providing a sterile, long term stable, hypoallergenic liquid human milk fortiﬁer that can be used in combination with human breast milk or infant formula without signiﬁcant volume displacement. The liquid human milk fortiﬁers of the present sure not only provide the signiﬁcant beneﬁt of ed sterility as compared to powdered human milk fortiﬁers that may potentially be subject to microbial contamination, but also provide a stable human milk fortiﬁer that includes an extensively hydrolyzed protein that can be more easily digested and absorbed into the gut of an , and particularly a preterm infant, as compared to intact proteins. The previous problems of providing a long term stable liquid human milk er with extensively hydrolyzed proteins has been overcome by including an OSA modiﬁed waxy potato starch in the human milk fortiﬁers disclosed herein. Even though extensively hydrolyzed proteins are well—known to have poor ability to form and stabilize emulsions and are highly prone to unwanted separation and sedimentation, the OSA modiﬁed waxy potato starch, in ation with extensively hydrolyzed protein, provides a long term stable emulsion with excellent sedimentation properties.

The use of OSA modiﬁed waxy potato starch es for improved l suspension in combination with improved emulsion stability in the liquid human milk fortiﬁer due to its unique physiochemical characteristics. Surprisingly, when the OSA modiﬁed waxy potato starch is introduced into the liquid human milk fortiﬁer, the concentrated liquid can be prepared with high levels of ble calcium salts and extensively hydrolyzed protein without mineral t or separation.

By providing a long term stable concentrated liquid human milk fortiﬁer that can bebased partly or solely on an extensively hydrolyzed protein ent, the t disclosure now provides a sterile, concentrated liquid product that can be used in neonatal ive care units in combination with human breast milk or infant fermula for preterm and term infants to provide the infant with the additional nutrients, including both protein and minerals, required for growth and maturation. This can now be done with a highly sterile, stable product that is highly concentrated so as tobe more like a powdered human milk fortiﬁer from a volume displacement . ctive.

Additionally, the use of the OSA modiﬁed waxy potato starch as a single - functional ingredient that provides for both emulsiﬁcation and stabilization allows for the use of a label friendly starch in many commercial markets around the world to provide enhanced physical stability in a concentrated liquid human milk fortiﬁer. This is a signiﬁcant advancement as many gum-type stabilizers, including eenan, are not favored in many countries around the World and alternatives should be used.

These and other optional ts or limitations of the concentrated s and methods of the present disclosure are described in detail hereinafter.

The terms "retort packaging" and "retort sterilizing" are used interchangeably herein, and unless otherwise speciﬁed, refer to the common practiCe of ﬁlling a container, most typically a metal can or other similar package, with a nutritional liquid and then ting the liquid-ﬁlled package to the necessary heat sterilization step, to form a sterilized, retort packaged, nutritional liquid product.

The term "aseptic packaging" as used herein, unless otherwise speciﬁed, refers to the manufacture of a packaged product without reliance upon the above—described retort W0 2013/,101401 . packaging step, wherein the nutritional liquid and e are ized separately prior to ﬁlling, and then are combined under sterilized or aseptic sing conditions to form a ized, aseptically packaged, nutritional liquid product.

The term "infan " as used herein, refers generally to individuals less than about 1 year of age, actual or corrected.

The term "preterm" are used herein refers to those infants born at less than 37 weeks gestation, having a birth weight of less than 2500 gm, or both.

The terms "fortiﬁer solids" or "total solids", unless otherwise speciﬁed, are used interchangeably herein and refer to all material components of the compositions of the present disclosure, less water.

The term “hypoallergenic” as used herein means that the concentrated liquid has a decreased tendency toprovoke an allergic reaction in a user, such as a preterm or term infant, as compared to non-hypoallergenic liquids. More particularly, the concentrated liquid is lergenic when there is 95% conﬁdence that 90% of allergic infants would not react to the liquid in a double-blind, placebo—controlled study (DBPC). An e of a suitable DBPC study is described in Kleinman, et 211. “Use of infant formulas in infants with cow milk allergy: a - review and recommendations,” Pediatr Allergy Immunol 1991, 4: 146-155.

The term “extensively hydrolyzed’ as used herein refers to protein that has been enzymatically or acidically hydrolyzed to have a degree of hydrolysis of at least 20%. Typically extensively yzed proteins exist primarily as di- and tripeptides.

The terms “liquid nutritional composition” and “nutritional liquid” are used interchangeably herein, and unless otherwise ed, refer to nutritional products, including human milk fortiﬁers, in trated form.

The terms e” and “shelf stable” as used herein mean that the concentrated liquid is resistant to separation of the liquid into two or more discernable layers (e. g., atop cream layer and a bottom serum layer) and precipitation/sediment formation on the bottom of a container for a time period after manufacture of at least three months desirably at least six ‘ months, desirably at least twelve months and more desirably at least 18 months.

All percentages, parts and ratios as used herein, are by weight of the total composition, unless otherwise speciﬁed. All such weights as they pertain‘to listed ingredients are based on the active level and, therefore, do not e solvents or by-products that may be - included in commercially available materials, unless otherwise speciﬁed.

Numerical ranges as used herein are ed to e every number and subset of numbers within that range, whether speciﬁcally disclosed or not. Further, these numerical ranges should be construed as providing support for a claim directed to any number or subset of numbers in that range. For example, a disclosure of from 1 to 10 should be construed as supporting a range of from 2 to 8, from 3 to 7, from 5 to 6, from 1 to 9, from 3.6 to 4.6, from 3.5 to 9.9, and so forth.

All references to singular characteristics or limitations'of the present disclosure shall include the ponding plural characteristic or limitation, and vice versa, unless otherwise speciﬁed or clearly implied to the contrary by the context in which the nce is made.

All combinations of method or s steps as used herein can be performed in any order, unless otherwise speciﬁed or Clearly implied to the contrary by the context in which the referenced combination‘is made.

The various embodiments of the concentrated liquids of the present disclosure may also be substantially free of any al or selected ingredient or e described , provided that the remaining concentrated] liquid still contains all of the required ingredients or features as described herein. In this context, and unless otherwise ed, the term “substantially free” means that the selected trated liquid contains less than a functional amount of the optional ingredient, typically less than 0.1% by weight, and also including zero percent by weight of such optional or selected ingredient.

W0 2013(101401 The concentrated s and corresponding manufacturing methods of the presentdisclosure can comprise, t of, or consist essentially of the elements and limitations of the disclosure as described herein, as well as any onal or optional ingredients, ‘ components, or limitations bed herein or otherwise useful in the concentrated .

Product Form While discussed ily as a concentrated liquid human milk fortiﬁer, it should be recognized by one skilled in the art that the concentrated itions described herein additionally include other concentrated liquid nutritional compositions that may be suspensions, emulsions or clear or substantially clear liquids. The resulting concentrated liquid nutritional compositions may be used as infant, pediatric, toddler, and/or adult liquid nutritional compositions and/or medicinal liquid nutritional compositions.

The concentrated liquid human milk ers of the present disclosure have a solids content of at least 10%, or even at least 20%, including from abOut 20% to about 55%, and including from about 20% to about 50%, and including from about 20% to about 45%, and including from about 20% to about 40%, and including from about 25% to about 40%, and including from about 29% to about 32% by weight. The concentrated liquid human milk ers are liquids that are capable of being poured directly from a package containing them into human milk or infant formula.

The concentrated liquid human milk fortiﬁers are generally formulated to have a caloric density of at least 1.25 kcal/ml (37 kcal/ﬂ oz),including from about 1.4 kcal/ml (42 kcal/ﬂ oz) to about 5 kcal/ml (149 kcal/ﬂ oz), and also including from about 1.5 kcal/ml (44 kcal/ﬂ oz) to about 2.5 kcal/ml (74 kcal/ﬂ oz), and also including from about 1.9 kcal/ml (56 kcal/ﬂ oz) to about 2.0 kcal/ml (59 kcal/ﬂ oz).

The trated liquid human milk fortiﬁers include packaged compositions further comprising a suitable unit dose package or ner. The term "unit dose" as used herein refers to indiyidual, single-use, packages of concentrated liquid human milk fortiﬁer containing an amount of human milk fortiﬁer that can be used in a preparation of an infant feeding to provide ient human milk fortiﬁer to supplement human milk for immediate use, e.g., preferably Within 8-24 hours, more preferably Within 0-4 hours, of mixing with human milk.

The amount of fortiﬁed human milk prepared for a premature , for example, typically ranges from 25 ml to 150 ml a day. Consequently, a single unit dose is the appropriate amount of fortiﬁer solids to fortify a 25 ml preparation. Multiple packages can be used to prepare larger feeding volumes, especially for term infants.

The amount or volume of concentrated liquid human milk fortifier in each unit dose package includes those embodiments in which the package contains an amount suitable to prepare an infant's next feeding. These unit dose packages typically contain sufﬁcient er to provide from about 0.5 g to about 10 g of fortiﬁer solids, more typically from about 0.8 g to about 7.5 g of fortiﬁer , and even more typically from about 0.85 g to about 6.0 g, of ier solids. [005 5] The concentrated liquid human milk fortiﬁers of the present disclosure are preferably formulated so as to provide fortiﬁed humanmilk having an osmolality of less than 500 mOsm/kg water, preferably from about 300 mOsm/kg water to about 400 mOsm/kg water.

Based on the disclosure herein, one skilled in the art can readily formulate the concentrated liquid human milk fortiﬁer with the riate carbohydrate sources‘and corresponding DE (dextrose lence) values to obtain or otherwise provide for the targeted osmolality of the human milk fortiﬁer When combined with human milk.

The concentrated liquid human milk ers may suitably have a pH ranging from about 3.5 to about 8.0, but are most advantageously in a pH range of from about 4.5 to about 7.5, including from about 4.5 to about 7.0, including from about 4.5 to about 6.7, including from about 4.5 to about 6.5, and including from about 4.5 to about 6.0. In some embodiments, the pH range is from about 5.5 to about 7.3, including from about 5.5 to about 7.0, including from about 5.5 to about 6.5, and further ing from about 5.5 to about 6.0.‘ In other ‘ embodiments, the pH range is from about 6.2 to about 7.2, including from about 6.2 to about 7.0, and including from about 6.2 to about 6.5. :12— Extensively Hydrolyzed Casein Protein The concentrated liquid human milk fortiﬁers of the present disclosure include hypoallergenic ively hydrolyzed casein as a protein source. Generally, the concentrated liquid human milk fortiﬁers will include at least 35%, including at least 50%, including at least 60%, including at least 75%, including at least 90%, and further including about 100% extensively hydrolyzed casein, by total weight of protein in the concentrated liquid human milk fortiﬁer. In one desirable embodiment of the present disclosure, the trated liquid human milk er includes 100% extensively hydrolyzed casein, by total weight of the n in the concentrated liquid human milk fortiﬁer. In this desirable embodiment, the concentrated liquid human milk fortiﬁer is hypoallergenic. In some embodiments, the concentrated liquid human milk fortiﬁer will include from about 35% to 100%, ing from about 50% to 100%, further including from about 75% to 100% extensively hydrolyzed casein, by total weight of protein in the concentrated liquid human milk fortiﬁer. As discussed further below, in some embodiments of the present disclosure, the concentrated liquid human milk ers of the present disclosure may optionally include other hypoallergenic or non-hypoallergenic proteins in addition to the extensively hydrolyzed casein protein. [005 8'] Extensively hydrolyzed casein proteins suitable for use in the Concentrated liquid human milk fortiﬁers of the present disclosure include those having a degree of hydrolysis of from about 5% to about 80%, including from about 20% to about 60%, and further including from about 40% to about 60%. Generally, the ively hydrolyzed casein has a ratio of total amino en (AN) to total nitrogen (TN) of from about 0.2 AN to 1.0 TN to about 0.4 AN to about 0.8 TN. Suitable commercially available extensively hydrolyzed caseins will generally have a protein level in the ient of from about 50% to about 95%, including from about 70% to about 90%. One le commercially available extensively hydrolyzed casein is Dellac CE90, which is a spray dried powder casein hydrolysate (Friesland Campina Domo, Amersfoort, the Netherlands).

. PCT/U82012/067589 Stabilizer System The concentrated liquid human milk fortiﬁers of the present disclosure include an octenyl succinic ide (0SA) modiﬁed waxy starch as a stabilizer, and in some embodiments, as the sole stabilizer present in the human milk fortiﬁer. As used herein, the term “waxy” is generally used to refer to es to starches that have a high level of amylopectin and a low level of amylose. In some embodiments, the amylopectin content of the “waxy” starch will be more than 90% by weight, including more than 95% by weight, including mOre than 99% by weight. In some embodiments, the waxy potato starches described in the present disclosure will only contain amylopectin and no amylose.

The OSA modified starch, including the desirable OSA modiﬁed waxy potato starch, is generally ed by esterifying a dextrinized, ungelatinized waxy potato starch with anhydrous octenyl succinic acid under alkaline conditions. Methods of this type are well known in the art. One suitable commercially available OSA modiﬁed waxy potato starch is ELIANETM MC 160 (AVEBE, The Netherlands).

Suitable OSA modiﬁed waxy potato starches for use in the liquid human milk fortiﬁers have an average granule size of from about 15 um to about 100 um, desirably from about 20 pm to about 100 um, desirably from about 30 am to about 100 um, which is generally larger than other modiﬁed starches, such as modiﬁed corn starch. The use of large granule—sized OSA modiﬁed waxy potato starches may potentially create strong, cohesive, and elastic networks improving the stabilization of concentrated liquids.

Further, le OSA modiﬁed waxy potato starches will have a orous t, t as phosphate monoesters, of from about 800 ppm to about 1000 ppm, which is lly higher than other es commercially available. For example, many corn es have a phosphorous content of about 30 ppm, which is signiﬁcantly below the desirable phosphorus content of the suitable dSA modiﬁed waxy potato starches. The phosphate monoesters have negatively charged molecules, which may increase the charge repulsion at the oil and water interface in the concentrated liquid human milk fortiﬁer, ting the oil droplets from coalescing and aggregating. Further, the increased charge repulsion causes an increase in water binding capacity, swelling powder, and Viscoelastic properties of the system. This may provide the concentrated liquid human milk fortiﬁers with an improved l suspension.

The OSA modiﬁed waxy starch, including the desirable OSA d waxy potato starch as described herein, is present in the concentrated liquid human milk fortiﬁerin an amount of from about 0.1% to about 3.5%, including from about 0.6%to about 2.0%, including from about 0.8% to about 1.5%, and further including about 1.2% by weight of the concentrated liquid human milk fortiﬁer.

In some embodiments, the trated liquid human milk fortiﬁer may include the OSA modiﬁed waxy potato starch in combination with one or more other izers including, for e, carrageenan or other gum—based stabilizers such as xanthan gum. In another embodiment, the OSA modiﬁed waxy potato starch is the sole stabilizer. In this ment, the er may be ntially free of carrageenan, or even completely free of carrageenan to allow a concentrated liquid human milk er to be prepared that does not include any carrageenan.

Macronutrients The concentrated liquid human milk fortiﬁers and concentrated liquid nutritional compositions of the present disclosure may comprise carbohydrate, fat, and protein macronutrients of sufficient types and amounts that, when used in combination with human milk or other feeding formula, help meet the nutritional needs of the user, especially the premature infant. In some desirable embodiments, the liquid concentrated human milk er will include a carbohydrate, protein, and fat. The concentration of these macronutrients in the various embodiments of the present disclosure includes the ranges described hereinafter.

Protein The concentrated liquid human milkfortiﬁers of the present disclosure comprise a protein suitable for use in infants, especially m infants, at concentrations ranging from about 1% to about 50%, ing from about 20% to about 40%, also including from about 5% to about 30%, including from about 10% to about 25%, and including from about 15% to about %, on a dry weight basis. In some desirable ments, the protein concentration may be from about 7’to about 15 grams, including from about 9 to about 12 grams of protein per 100 grams of ﬁnal liquid product.

As noted above, the protein component of the concentrated liquid human milk fortiﬁers of the present disclosure is at least partially comprised of extensively hydrolyzed casein. In a particularly desirable embodiment of the present disclosure, the protein component of the concentrated liquid human milk fortiﬁer is entirely comprised of extensively hydrolyzed casein. In embodiments wherein additional proteins sources (i.e., one or more n sources in ' addition to the extensively hydrolyzed protein source) are to be used in the concentrated liquid human milk fortiﬁer in addition to the extensively hydrolyzed casein (i.e., the concentrated liquid human milk fortiﬁer protein component is not 100% extensively hydrolyzed casein), the fortiﬁer may still be made hypoallergenic by including additional lergenic proteins such as soy protein hydrolysate, whey protein hydrolysate, rice protein hydrolysate, potato protein hydrolysate, ﬁsh protein hydrolysate, egg albumen hydrolysate, gelatin n hydrolysate, pea protein hydrolysate, bean protein hydrolysate, combinations of animal and vegetable protein ysates, and combinations thereof.

In this context, the terms "protein hydrolysates" or "hydrolyzed protein" are used interchangeably herein and include ively yzed proteins, wherein the degree of hydrolysis is most often at least 5%, including from about 10% to about 80%, and also including from about 30% to about 80%, even more preferably from about 40% to about 60%. The degree of hydrolysis is the extent to which peptide bonds are broken by a ysis method. The degree ofprotein hydrOlysis for es of characterizingthe extensively hydrolyzed protein component of these embodiments is easily ined by one of ordinary skill in the formulation arts by quantifying the amino nitrogen to total nitrogen ratio (AN/TN) of the protein component of the Selected formulation. The amino en component is quantiﬁed by USP ion methods for determining amino nitrogen content, while the total nitrogen component is -16— determined by the Tecator Kjeldahl method, all of which are well known methods to one of ordinary skill in the analytical chemistry art, In other embodiments of the present disclosure, the concentrated liquid human milk fortiﬁer, in addition to the extensively hydrolyzed n, may include an additional non— hypoallergenic protein source including for example, partially hydrolyzed or non-hydrolyzed (intact) protein, and can be d from any known/or ise suitable source such as milk (e.g., casein, whey, lactose-free milk protein isolates), animal (e.g., meat, ﬁsh), cereal (e.g., rice, corn), vegetable (e.g., soy, pea, bean), Or combinations thereof. The protein can include, or be entirely or partially replaced by, free amino acids known or otherwise suitable for use in nutritional ts, non-limiting examples of which include L-alanine, L-arginine, L- asparagine, L-aspartic acid, L—carnitine, L-cystine, L—glutamic acid, L—glutamine, glycine, L— histidine, L-isoleucine, L-leucine, L—lysine, L-methionine, ylalanine, L—proline, L-serine, L—taurine, L—threonine, tophan, L-tyrosine, L-valine, and combinations thereof.

Carbohydrate The concentrated liquid human milk iers of the present disclosure comprise a carbohydrate suitable for use in infants, especially preterm infants, at concentrations most lly ranging up to about 75% by weight on a dry weight basis, including from about 5% to about 50%, and also including from about 20% to about 40%, by weight on a dry weight basis.

Carbohydrates suitable for use in the concentrated liquid human milk iers may include maltodextrin (i.e., non—sweet, nutritive polysaccharide having a DE value less than ), corn maltodextrin, glucose polymers, sucrose, corn syrup, corn syrup solids (i.e., polysaccharide having a DE value greater than 20), glucose, rice syrup, fructose, high se corn syrup, indigestible oligosaccharides such as fructooligosaccharides (F08), and combinations thereof. The carbohydrates may comprise lactose or can be substantially free of lactose. One particularly red carbohydrate is maltodextrin. [0072'] One embodiment ofthe present disclosure includes a non-reducing ydrate component, which may represent from about 10% to 100%, including from about 80% to 100%, and also ing 100%, by weight of the total carbohydrate in the trated liquid human milk fortiﬁer. The ion of a non—reducing Carbohydrate may enhance the product stability and is generally better tolerated by infants, especially premature infants. Non-limiting examples of non-reducing carbohydrates e sucrose or other carbohydrates that do not readily oxidize or react with Tollen's, Benedict's, or Fehling‘s reagents. The present disclosure therefore includes those embodiments sing a carbohydrate component, wherein the carbohydrate component comprises a mono— and/or disaccharide such that at least 50%, including from about 80% to 100%, and also including 100%, of the mono- and/or disaccharide is a non—reducing carbohydrate.

F__a_t The concentrated liquid human milk fortiﬁers of the present disclosure also comprise a fat component suitable for use in infants, especially preterm infants, at concentrations most typically ranging up to about 75% by weight on a dry weight basis, including from about % to about 50%, and also including from about 20% to about 40%, by weight on a dry weight basis.

Fats suitable for use in the concentrated liquid human milk fortiﬁers of the present disclosure may includecoconut oil, soy oil, corn oil, olive oil, safﬂower oil, high oleic safﬂower oil, MCT oil (medium chain triglycerides), sunﬂower oil,lhigh oleic sunﬂower oil, structured triglycerides, palm and palm kernel oils, palm olein, canola oil, marine oils, seed oils, and combinations thereof. Particularly preferred fats include MCT oil, soy oil, and coconut oil, which may be used alone or in any combination.

Suitable fats for use in the concentrated liquid human milk ers include emulsiﬁers to help the various fortiﬁer components readily disperse when ed with human milk. Non-limiting examples of suitable emulsiﬁers include glyceryl monostearate, monoglycerides, erides, distilled ycerides, soya bean lecithin, ythylene stearate, polyoxyethylene sorbitan mono-oleate, polyoxyethylene sorbitan monopalmitate, polyoxyethylene an monostearate, ammonium phosphatides, polyoxyethylene sorbitan WO 01401 PCT/U82012/067589 . monolaurate, citric acid esters of mono and diglycerides of fatty acids, tartaric acid esters of mono and diglycerides of fatty acids, and combinations thereof. Natural soy lecithin is» especially useful in this respect.

The fat component of the concentrated liquid human milk fortiﬁer may therefore optionally include any emulsiﬁer suitable for use in infant nutritional products. Emulsiﬁer concentrations in these products may range up to about 10%, including from about 1% to about %, even more typically from about 1.5% to about 5%, by weight of the total fat component. In one embodiment, the emulsiﬁer is in the product in an amount of about 2%, by weight of the total fat ent.

The concentrated liquid human milk fortiﬁers of the present disClosure also , include those embodiments that comprise as part of the fat component one or more of arachidonic acid, docosahexaenoic acid, or combinations thereof, alone or in further combination with linoleic acid, linolenic acid, or both. [007 8] Without being limiting, one suitable formulation of macronutrients for the concentrated liquid human milk fortiﬁers of the present disclosure is set forth in the table below.

Grams per 100 Grams Kcal per nutrients ier a ate %) I 7 Protein 9.2 28.42 ' l Fat | 4.3 29.88 Carbohydrate 1 13.5 | 41.7 .

Ash | . 2.74‘ | -— Total Solids | 30 | -- Kcal ‘ | 129.5 | -- Vitamins and Minerals The concentrated liquid human milk fortiﬁers of the present disclosure may further se any of a variety of Vitamins, nOn-limiting examples of which include vitamin A, n D, vitamin E, vitamin K, thiamine, riboﬂavin, pyridoxine, vitamin B12, niacin, folic acid, pantothenic acid, biotin, Vitamin C, choline, inositol, salts and derivatives thereof, and combinations thereof.

The trated liquid human milk fortiﬁers may also further comprise any of a variety of minerals known or otherwise suitable for use in infant or other nutritional formulas, non-limiting examples of which include phosphorus, magnesium, calcium, zinc, manganese, copper, iodine, sodium, potassium, chloride, selenium, and combinations thereof.

The concentrated liquid human milk fortiﬁers of the present disclosure include those embodiments sing per 100 kcal of er solids one or more of the following: Vitamin A (from about 250 to about 6500 IU), vitamin D (from about 40 to about 1200 IU), vitamin K, n E (at least 0.3 IU), Vitamin C (at least 8 mg), thiamine, n B12, niacin, folic acid, pantothenic acid, biotin, choline (at least 7 mg), and inositol (at least 2 mg).

The concentrated liquid human milk fortiﬁers also include those embodiments comprising per 100 kcal of the fortiﬁer solids one or more of the following: calcium (at least 50 mg), phosphorus (at least 25 mg), ium (at least 6 mg), iodine, zinc (at least 0.5 mg), copper, manganese, sodium (from about 20 to about 60 mg), ium (from about 80 to about 200 mg), chloride (from about 55 to about 150 mg) and selenium (at least 0.5 mcg).

Other Optional Ingredients The concentrated liquid human milk fortiﬁers of the present disclosure may further optionally se other ingredients that may modify the physical, chemical, aesthetic of processing characteristics of the compositions or serve as pharmaceutical or additional such optional ients . nutritional components when used in the targeted population. Many are known for use in food and nutritional ts, including infant formulas, and may also be used in the concentrated liquid human milk fortiﬁers of the present disclosure, provided that such optional materials are compatible with the materials described herein, are safe and effective for their intended use, and do not otherwise unduly impair t performance.

Non-limiting examples of such al ingredients e preservatives, anti- oxidants, various pharmaceuticals, buffers, carotenoids, colorants, ﬂavors, nucleotides and sides, thickening agents, prebiotics, sialic acid-containing materials, and other excipients or processing aids.

Aseptic ing The concentrated liquid human milk fortiﬁers of the present disclosure may be sterilized and aseptically packaged. The aseptic packaging can be accomplished using any of a variety of techniques well known to those of ry skill in the formulation art, so long as the heat treatment is sufﬁcient to achieve long term shelf stability of the concentrated liquid. In one speciﬁc example, an aseptic process is utilized that includes a high temperature short time (HTST) processing step (i.e., about 165°F (74°C) for about 16 s) or an ultra high temperature (UHT) processing step (i.e., about 292°F (133°C) for about 5 seconds).

A typical aseptic prOceSs in accordance with the present disclosure involves the preparation of a slurry from one or more ﬂuid ations that may contain water and one or more of the following: carbohydrates, OSA modiﬁed waxy potato starch, ively hydrolyzed casein protein, fats, vitamins and minerals. This slurry is typically emulsiﬁed, deaerated, homogenized and cooled to form a sterilized a, and then aseptically packaged to form a sterilized, aseptically packaged concentrated liquid human milk fortiﬁer.‘ Various other solutions may be added to the slurry at most any time before, during, or after processing.

Suitable aseptic packaging techniques include any of the well known aseptic packaging methods‘disclosed in the formulation arts for preparing nutritional formulation, all of which are lly directed to the sealing or ﬁlling of a sterilized liquid into a sterilized, air- tight container. Many variations on icmethod exist and are well known to those of ordinary skill in the formulation art, non—limiting examples of which are described in US. Pat.

‘ No. 6,096,358 (Murdick et al.); US. Pat. No. 6,227,261 (Das et al.); and US. Pat. No. 6,371,319 n et a1), which descriptions are incorporated herein by reference.

The aseptically packaged embodiments ofthe t disclosure may include any container or package suitable for use with liquid human milk fortiﬁers and also capable of withstandingaseptic processing conditions (e.g., high temperature sterilization). Non-limiting examples of such containers include single or multi use bags, plastic bottles or containers, s, metal cans glass bottles, foil or other ﬂexible pouches, syringes, vials, or any other container meeting the above—described criteria.

The aseptically packaged container for these embodiments is typically sterilized prior to being ﬁlled with its ized contents. The container is most typically ized by the ation of hydrogen peroxide or other suitable disinfectant to the inside surface of the container. The hydrogen peroxide or other ectant is often applied in an atomized mist.

After a disinfeCtant is applied, the container may be transported along a conveyor system during which time the container may be subjected to one or more sprayings of hot sterilized air, preferably hot, sterilized, dry air. The container is then preferably injected with nitrogen gas.

The aseptically prepared ner is then aseptically ﬁlled with sterilized product and sealed.

For aseptic packaging, the concentrated liquid human milk fortiﬁer is typically heat treated with a high temperature short time (HTST) process or an ultra high temperature (UHT) process to sufﬁciently reduce the bioburden to allow the products to be commercially sterile over an extended shelf life of the ﬁnished product exceeding about 12 months. The treated formula is then homogenized at 1000 psi or higher and aseptically packaged.

In an ative ment, the concentrated liquid human milk fortiﬁers of the t disclosure may also 'be sterilized and retort ed utilizing conventional means known in the art.

Methods of Use The concentrated liquid human milk fortiﬁer of the t disclosure is used in combination with human milk or other suitable infant formula, wherein the resulting fortiﬁed has an osmolality suitable for oral stration to an . human milk or fortiﬁed infant formula infant. As noted, the osmolality will most typically be less than about 500 g water, more typically from about 300 mOsm/kg water to about 400 mOsm/kg water.

The concentrated liquid human milk fortiﬁer of the present disclosure may be added directly to human milk or to infant formula in a volume to volume ratio of from about 1:3 to about 1:10, including from about 1:3 to about 1:9, ing from about 1:3 to about 1:8, including from about 1:3.5 to about 1:7, also including from about 1:4 to about 1:6, and also including from about 1:5 to about 1:6. The ratio is ultimately selected based primarily upon the ients and osmolality of the concentrated liquid human milk fortiﬁer and in view of the particular ional needs of the infant. The concentrated liquid human milk er may be added directly to every feeding or to a sufﬁcient number of feedings (e. g., once or twice daily) to provide optimal nutrition in View of the particular nutritional needs of the infant.

Human milk or other infant formula, after fortiﬁcation with the concentrated liquid human milk fortiﬁer will most typically have a caloric density g from about 19 kcal/ﬂ 02 (0.64 kcal/ml) to about 26.7 kcal/ﬁ oz (0.9 kcal/ml), with the 22—25 kcal/ﬂ oz formulations (0.74-0.84 kcal/ml) being more useful in preterm infants, and the 19—21 kcal/ﬂ oz ’ (0.64—0.71 kcal/ml) ations more useful. for term infants.

The s ofthe present disclosure include methods of providing nutrition to infants, and especially preterm infants. As noted herein, preterm infants may especially beneﬁt from the use of human milk fortiﬁers as the fortiﬁers can provide additional nutrients to the m infant when combined with human breast milk and/or infant a to foster quicker growth and development. In one particular embodiment, nutrition is provided to an infant by the addition of the concentrated liquid human milk fortiﬁer to human breast milk or infant formula or combination thereof followed by the administration of the fortiﬁed human breast milk or infant formula to the infant.

Other alternative methods ofthe present disclosure include using the human milk fortiﬁers as described herein to fortify human breast milk, infant formula, or a combination of human breast milk and infant formula to provide a fortiﬁed nutritional liquid for administration to an infant, and particularly a preterm infant. In one embodiment, human breast milk is fortiﬁed by mixing the concentrated human milk fortiﬁer with human breast milk or infant formula or a combination thereof in a volume to volume ratio of from about 1:3 to about 1:10.

The methods of the present disclosure also include a method of providing nutrition to users other than infants, such as adults and elderly. This method includes the addition of the concentrated composition to other liquid nutritionals such as suspensions, emulsions or clear or substantially clear liquids. The resulting concentrated liquid nutritional compositions may be used as adult liquid nutritional compositions and medicinal liquid nutritional compositions.

Methods of Manufacture The concentrated liquid human milk fortiﬁers of the present disclosure may be , prepared in accordance with the methods described hereinafter.

In one embodiment, the concentrated liquid human milk fortiﬁer is prepared by solubilizing and ing/mixing ingredients into a homogeneous aqueous mixture which is subjected to a sufﬁcient thermal treatment and aseptic ﬁlling to achieve long term physical and ial shelf stability.

To begin the manufacturing s, macronutrients (carbohydrate, protein, fat, and minerals) are combined in l es together and with water. This blend is‘ subjected to an initial heat treatment and then tested to verify proper nt levels. onal detail on this process is provided in the following aphs.

An intermediate aqueous carbohydrate-mineral (CHO-MIN) slurry is prepared by heating an appropriate amount of water. With agitation, the following soluble ingredients are added: maltodextrin, ium citrate, magnesium chloride, potassium de, sodium chloride, and choline chloride. The carbohydrate—mineral slurry is held’at elevated temperature under agitation until added to the blend.’ 2012/067589 An intermediate oil slurry is prepared by heating MCT oil and coconut oil to an elevated temperature and dding distilled monoglycerides with agitation for a minimum of minutes in order for the ingredient to ve. Soy oil, vitamin A palmitate, vitamin D3, di- alpha-tocopheryl—acetate, phylloquinone, ARA, DHA, and mixed carotenoids are then added with agitation to the oil blend. Insoluble mineral calcium source and ultra micronized tricalcium ate are added to the oil. OSA modified waxy potato starch is then added to the oil blend with proper agitation. The oil blend slurry is maintained at an elevated temperature under agitation until added to the blend.

The blend is prepared by combining the ingredient water, casein hydrolysate, all of the N slurry and whole oil blend slurry. The blend is maintained at 120°F for a period of time not to exceed two hours before further processing.

The blend is then homogenized using one or more in-line homogenizers at pressures from 1000-4000 psig with or without a second stage nization from 0 psig followed by heat treatment using a UHTST (ultra-high temperature short time, 292-297°F for 1-30 seconds) process. After the appropriate heat treatment, the batch is cooled in a plate cooler to 33445°F and then transferred to a refrigerated holding tank, where it is subjected to ical testing.

The next step in the manufacturing s involves adding any desired vitamins, trace minerals and water in order to reach the ﬁnal target total solids and vitamin/mineral contents. The ﬁnal batch is ﬁlled into a suitable container under aseptic conditions or treated with a terminal ization process so the product will be stable at room temperature for an _ extended shelf life. onal detail on this process is provided in the following paragraphs.

A trace mineral/vitamin/nutrient on (STDI) is prepared by heating water to 80—100°F and adding the following ingredients with agitatiOn: potassium citrate, ferrous sulfate, zinc sulfate, copper sulfate, manganese sulfate, sodium selenate, pyridoxine hydrochloride, riboﬂavin, thiamine hydrochloride, cyanocobalamin, folic acid, calcium pantothenate, niacinamide, biotin, m-inositol, nucleotide/choline premix, L-camitine, L—leucine, L-tryptophan, and L-tyrosine.

A vitamin C solution (STDZ) is prepared by adding ascorbic acid to water solution with agitation.

Both STDl and STD2 solutions are then added to the refrigerated batch, with agitation. The appropriate amount of ingredient dilution water is then added to the batch to achieve a target total solids level of at least 5%, including at least 10%, and preferably about 20- 55%. The ﬁnal batch is then subjected to appropriate thermal treatment and ﬁlled into a suitable container under aseptic conditions and ses.

The concentrated liquid human milk fortiﬁers of the present disclosure may, of course, be manufactured by other known or ise suitable techniques not speciﬁcally described or shown herein without ing from the spirit and scope of the present disclosure.

The present embodiments are, therefore, to be ered in all respects as rative andnot restrictive and that all changes and? equivalents also come within the description of the present disclosure. The following non—limiting examples will further illustrate the formulations and methods ofthe present sure.

EXAMPLES The following examples illustrate speciﬁc embodiments and/or features of the concentrated liquid human milk ers of the present disclosure. The examples are given solely for the purpose of illustration and are not to be construed as limitations of the present disclosure, as many variations thereof are possible without ing from the spirit and scope of the disclosure. All exempliﬁed s are weight percentages based upon the total weight of the formulation, unless ise speciﬁed.

Examples 1—4 In Examples 1—4, concentrated liquid human milk fortiﬁers may be prepared in accordance with the present disclosure. The ingredients for the concentrated liquid human milk er are shown in the f0110wing table. All ingredient amounts are listed as pound per approximately 1000 pound batch of product, unless otherwise speciﬁed. ' 2012/067589 Ingredient (Per Example 1 Example 2 Example 3 Example 4 ient Water q.s. I q.s. q.s.

I Casein Hydrolysate 110.5 110.5 . 110.5 110.5 Maltodextrin 110.5 110.5 ' ‘ 110.5 110.5 MCT Oil 18.9 ‘ 18.9 18.9 18.9 Tricalcium Phosphate 14.8 14.8 14.8 14.8 OSA Modiﬁed Waxy 12.0 10.8 9.6 13.2 ' Potato Starch ’ Soinl 11.3 11.3 I 11.3 11.3 Coconut Oil ' . 6.9 6.9 I 6.9 6.9 Potassium e 5.2 5.2 I 5.2 5.2 Magnesium Chloride . 3.4 3.4 3.4 3.4 Ascorbic Acid 3.3 3.3 3.3 3.3 I M. Alpina Oil 2.6 2.6 . 2.6 2.6‘ I c. Cohnii 011 2.1 2.1 2.1 2.1 Leucine 1.84 I 1.8 1.8 1.8 Potassium Chloride 1.7 I 1.7 1.7 1.7 Tyrosine 1.4 1.4 1.4 1.4 Monoglycerides 390.1 g 390.1 g I 390.1 g I_390.1 g Sodium Chloride 345.3 g I 345.3 g 345.3 g I M-Inositol 200.0 g 2000 g Choline Chloride . 160.0 g Tryptophan 150.0 g Zinc Sulfate Niacinamide 127.3 g 127.3 g . . di-Alpha—Tocopheryl , 108.7 g 108.7 g Acetate . 100.0 g . 60-0 g 60.0 g Ferrous Sulfate . 52 0 g I 52 0 g 52.0 g Vitamin A Palmitate 25.8 g 25.8 g Riboﬂavin 15.0 g 15.0 g Thiamin Hydrochloride 11.0 g Pyridoxine 8.9 g . 8.9 g Hydrochloride Cupric Sulfate 6.0 g Vitamin D3 Folic Acid Beta Carotene ' 940.0 mg Biotin 860.0 mg Manganese e 840.0 mg Phylloquinone 285.0 mg Sodium Selenate 44.0 mg Cyanocobalamin 40.0 mg Potassium Hydroxide q.s. ~28- The concentrated liquid human milk fortiﬁer is prepared by solubilizing and combining/mixing ingredients into a homogeneous aqueous e which is subjected to a sufﬁcient thermal treatment and aseptic ﬁlling to achieve long term physical and microbial shelf stability.

To begin the manufacturing process, macronutrients (carbohydrate, protein, fat, and minerals) are combined in several slurries together and with water. This blend is subjected to an initial heat treatment and then tested to verify proper nutrient levels. Additional detail on this process is provided in the following paragraphs.

An ediate aqueous carbohydrate-mineral (CHO-MIN) slurry is prepared by heating an appropriate amount ofwater to l40-l60°F. With agitation, the following soluble ingredients are added: inaltodextrin, potassium citrate, magnesium de, potassium chloride, sodium chloride, and choline chloride. The carbohydrate-mineral slurry is held at 130-150°F under agitation until added to the blend.

An intermediate oil slurry is prepared by g MCT oil and coconut oil to 150 to 170°F andthen adding distilled monoglycerides with agitation for a minimum of 10 minutes in order for the ingredient to dissolve. Soy oil, nA palmitate, ha—tocopheryl e, phylloquinone, vitamin D3, ARA-containing oil, DHA-containing oil, lutein, and arotene are then ith agitation to the oil blend. Insoluble mineral calcium source, and ultra micronized tricalcium phosphate is added to the oil. OSA d waxy potato starch is then added to the oil blend with proper agitation. The oil blend slurry is maintained at 130-150°F under agitation until added to the blend. [01 l6] The blend is prepared by combining'the ingredient water, casein hydrolysate, all of the CHO-MIN slurry and whole oil blend slurry. The blend is maintained at 120°F for a period oftime not to exceed two hours before further processing.

The blend is then homogenized using one or more in-line nizers at pressures from 1000—4000 psig with or without a second stage homogenization from [100-500 psig followed by heat ent using a HTST (high temperature short time, 165—185°F for 15-20 seconds) process. After the appropriate heat treatment, the batch is cooled in a plate cooler to 33-45°F and then transferred to a refrigerated g tank, Whereit is subjected to analytical testing.

The next step in the manufacturing process es adding vitamins, trace minerals, other ingredients, and water in order to reach the ﬁnal target total solids and vitamin/mineral contents. The ﬁnal pH of the product prior to thermal. treatment is also adjusted.

The ﬁnal batch is ﬁlled into a suitable container under aseptic conditions or treated with a terminal ization process so the product will be stable at room temperature for an extended shelf—life. Additional detail on this process is provided in the following paragraphs.

A trace mineral/Vitamin/nutrient solution (STDl) is prepared by heating water to 80-100°F and adding the following ingredients with agitation: potassium citrate, ferrous sulfate, zinc sulfate, copper sulfate, manganese sulfate, sodium selenate, xine hydrochloride, avin, thiamine hydrochloride, vitamin B12, folic acid, calcium pantothenate, amide, biotin, m—inositol, L—carnitine, leucine, and tyrosine.

A Vitamin C solution (STD2) is ed by adding ascorbic acid to a water solution with agitation.

All STD‘l and STD2 ons are then added to the refrigerated batch, with agitation. The appropriate amount of ingredient dilution water is then added to the batch to achieve a target total solids level of 20.0—55.0%. The ﬁnal pH of the product prior to thermal treatment is adjusted to >4.6—5.0 by addition of citric acid. Theﬁnal batch is then subjected to appropriate thermal treatment and ﬁlled into a suitable container under an aseptic conditions and pI‘OCCSSCS.

Example 5 In this Example, the stabilization teristics of OSA modiﬁed waxy potato starch in a concentrated liquid human milk fortiﬁer were analyzed and compared to a control -3 0- concentrated liquid human milk fortiﬁer without a stabilizer and a concentrated liquid human ‘ milk fortiﬁer using OSA modiﬁed waxy corn starch as a stabilizer.

Three s of liquid human milk fortiﬁer including extensively hydrolyzed casein were prepared using a method similar to the method described in es 1—4: A first sample was the control to which no stabilizer was added. A second sample was prepared ing 1.2% by weight OSA modiﬁed waxy corn starch (Uni-Pure® IMF 2332, Ingredion Incorporated (Westchester, IL) Starch Food tion, Bridgewater, New Jersey) as a izer.

A third sample was prepared including 1.2% by weight OSA d waxy potato starch (ELAINETM MC-160, AVEBE, The lands) as a stabilizer. The samples were then stored at least 6 months at room temperature.

As shown in the ﬁgures, the control sample ( showed heavy creaming, wherein oil and oil soluble nutrients separated into two different layers. Further, there was heavy sedimentation of insoluble minerals such as calcium. The second sample (, which included the OSA modiﬁed waxy corn starch, showed some creaming and phase separation.

Further, the second sample had heavy sedimentation due to insoluble minerals. The third sample (, which included ‘OSA modiﬁed waxy potato starch, showed no phase separation or mineral sedimentation (i.e.’, mineral fallout). This indicates that the use of OSA modiﬁed waxy potato starch as a stabilizer allowed for a stable trated liquid human milk fortiﬁer to be prepared to contain ively hydrolyzed casein and high levels of insoluble minerals without causing mineral fallout or defects in emulsion stability.

Further, the s including the three samples were turned upside down and the results are shewn in FIGS. 4A-4C. As shown in , the control sample shows heavy creaming and sedimentation. The second sample () including OSA modiﬁed waxy corn starch shows some creaming and sedimentation. The third sample () including OSA modiﬁed waxy potato starch shows little to no creaming along with little to no sedimentation.

Example 6 In this Example, concentrated liquid human milk fortiﬁers having varying stabilizers were analyzed for elasticity behavior as a function of ation (i.e., strain).

Three trated liquid human milk fortiﬁers as prepared in Example 5 were used for this Example. Strain sweep measurements for each sample were then obtained using an ARES G2 rheometer, ble from TA Instruments (New Castle, Delaware). The linear range in the resulting data corresponded to elastic modulus wherein change as a function of strain was insigniﬁcant. The line ﬁt was performed to identify the u elastic modulus. The frequency of the measurement was 10 rad/s. The results are shown in As shown in the sample ing OSA modified waxy potato starch demonstrates a long linear range with no ilities indicating stable emulsion and suspension characteristics. The sample including OSA modiﬁed waxy corn starch demonstrates break in the structure, indicating instabilities at low strains and having low elastic modula associated with a small linear range. This indicates that this sample does not have as stable of emulsion and suspension characteristics as compared to the sample ing OSA modiﬁed waxy potato starch. The control sample shows instabilities at low strains with break in the structure and minimal linear range, which is a sign ofpoor emulsion and suspension characteristics.

Example 7 'In this Example, concentrated liquid human milk fortiﬁers having varying stabilizers were analyzed for dynamic modula as a function of frequency and temperature.

The three concentrated liquid human milk fortiﬁers as prepared in Example 5 were used for this Example. Strain sweep ements for each sample were obtained as in e 6 as a ﬁmction of strain at room temperature. The frequency of the measurements ranged from about 4.8 rad/s to about 218.2 rad/s. The results are shown in 2012/067589 As shown in the dynamic modula, e.g., 88.6 mPa at 46.4 rad/s, for the control sample at low frequencies was low, indicating poor suspension stability. It is expected that this sample would be a Viscoelastic gel. Gel—like products typically e water out, causing phase tion. This sample’s strain sweep spectra also showed instabilities at low strains with very small linear range, further conﬁrming lack of emulsion and suspension stabilities.

The dynamic modula, e.g., 70.8 mPa at 46.4 rad/s, for the sample with OSA modiﬁed waxy corn starch at low frequencies was also low, indicating poor suspension stability.

The QSA modiﬁed waxy potato starch sample’s mechanical spectra demonstrates that the product is a lastic liquid having no gel-like or. Further, the low frequency elastic modula, e.g., 206.4 mPa at 46.4 rad/s, is signiﬁcantly larger than the other samples with no defects showing improved suspension stability. In addition, the high frequency elastic modula, e. g., 562.9 mPa at 184.8 rad/s, is signiﬁcantly larger than the other samples with no . defects indicating good emulsion characteristics. Both measurements of elastic modula as a function of frequency and strain sweep were aligned well, showing good emulsion and suspension characteristics for this sample.

WE

Claims

CLAIM :

1. A concentrated liquid human milk fortifier, comprising: from 1% to 50% by weight protein on a dry weight basis, octenyl succinic anhydride modified waxy potato , fat, vitamins and minerals, wherein at least a portion of the protein is extensively hydrolyzed casein.

2. The concentrated liquid human milk fortifier ing to claim 1, wherein 100% by weight of the protein is extensively hydrolyzed casein.

3. The trated liquid human milk fortifier according to either one of claims 1 or 2, comprising from 0.1% to 3.5% by weight l succinic anhydride modified waxy potato starch.

4. The concentrated liquid human milk fortifier according to any one of the preceding , wherein the concentrated liquid human milk fortifier has a solids content of at least 5% by weight.

5. The concentrated liquid human milk fortifier according to any one of the preceding claims, wherein the concentrated liquid human milk fortifier is an aseptically-sterilized concentrated liquid human milk fortifier.

6. The concentrated liquid human milk ier according to any one of the preceding claims, wherein octenyl ic anhydride modified waxy potato starch is a sole stabilizer of the concentrated liquid human milk fortifier.

7. The concentrated liquid human milk fortifier according to any one of the preceding claims, wherein the fortifier is eenan-free and comprises from 1% to 50% by weight of extensively hydrolyzed hypoallergenic casein, from 0.8% to 1.5% by weight, octenyl succinic anhydride modified waxy potato starch, and a solids content of from 20% to 55% by weight.

8. The concentrated liquid human milk fortifier according to claim 7, wherein octenyl succinic anhydride ed waxy potato starch is a sole stabilizer. (10143670_1):JJC

9. A concentrated liquid human milk fortifier according to any one of the preceding claims for use in fortifying human breast milk.

10. A method of fortifying breast milk, comprising: mixing a concentrated liquid human milk fortifier with breast milk in a volume to volume ratio of from 1:3 to 1:10, the concentrated liquid human milk fortifier comprising from 1% to 50% by weight protein, on a dry weight basis, and octenyl succinic anhydride modified waxy potato starch, wherein at least a portion of the protein is extensively hydrolyzed .

11. The method according to claim 10, wherein 100% by weight of the protein is extensively hydrolyzed casein.

12. The method according to either one of claims 10 or 11, wherein the extensively hydrolyzed casein is hypoallergenic protein.

13. The method according to any one of claims 10 to 12, wherein the concentrated liquid human milk fortifier ses from 0.1% to 3.5% by weight, octenyl succinic ide ed waxy potato starch.

14. The method ing to any one of claims 10 to 13, wherein the octenyl succinic anhydride modified waxy potato starch is a sole stabilizer.

15. The concentrated liquid human milk fortifier according to any one of claims 1 to 6, wherein the concentrated liquid human milk fortifier is carrageenan-free.

16. The concentrated liquid human milk fortifier according to any one of claims 1 to 8, wherein the extensively hydrolyzed casein is hypoallergenic protein.

17. The concentrated liquid human milk fortifier according to any one of claims 1 to 6, comprising from 0.8% to 1.5% by weight octenyl succinic anhydride ed waxy potato starch.

18. The concentrated liquid human milk fortifier according to any one of claims 1 to 6, wherein the concentrated liquid human milk ier has a solids t of from 20% to 55% by weight. (10143670_1):JJC

19. The method according to any one of claims 10 to 14, n the concentrated liquid human milk fortifier is mixed with breast milk in a volume to volume ratio of from 1:3.5 to 1:7.

20. The method according to any one of claims 10 to 14, wherein the concentrated liquid human milk fortifier comprises from 0.8% to 1.5% by weight octenyl succinic anhydride modified waxy potato starch.

21. Use of a concentrated liquid human milk fortifier according to any one of claims 1 to 8 and 15 to 18 for fortifying breast milk.

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