Human milk fortifier micronutrient supplementation kit

Abstract

The present invention relates to a human milk micronutrient supplementation kit, and methods of use thereof.

Description

HUMAN MILK FORTIFIER MICRONUTRIENT SUPPLEMENTATION KITField of the invention

[0001] The present invention relates to a human milk fortifier micronutrient supplementation kit, and methods of use thereof.Background to the invention

[0002] Preterm or premature infants are categorised as those infants born prior to the 37th week of gestation and / or weighing less than 2,500 grams at birth (low birth weight (LBW) infants). Preterm infants weighing less than 1 ,500 grams are classified as very low birth weight (VLBW) infants. Each year, approximately 50,000 neonates are classified as VLBW in the USA1.

[0003] Growth rates for preterm infants should ideally meet or exceed age-specific intrauterine growth rates in order to mimic rates comparable to term-born infants. Due to their underdeveloped digestive systems, lack of placental transfer of key nutrients such as iron, and overall developmental immaturity, preterm infants present special nutritional needs.

[0004] When a mother’s own milk (MOM) is unavailable or insufficient, donor human milk (DHM) is the recommended alternative. However, because of the immature digestive systems, increased nutrient needs and limited enteral volume tolerance, DHM is nutritionally insufficient to meet the demands of this population.

[0005] . Recommended enteral volume goals for the preterm population are 130-200 mL / kg / day, with an average of 150 mL / kg / day consumed once at full enteral feedings. Since the caloric content of human milk is approximately 67 kcal per 100 ml of milk (20 kcal per fluid ounce of milk), and preterm infants require approximately 120 kcal per kg of weight per day, the volume of human milk that can be tolerated by these infants supplies less than 80% of the infant’s energy needs. In addition, the protein demands of the VLBW infant are 4-4.5 g / kg / day. Preterm MOM provides approximately 1.6-2.2 g / dL protein, while DHM provides 1 g / dL. In either case, the amount of unfortified human milk needed to meet the protein demands of the VLBW infant exceeds the enteral feeding volume that is considered safe.

[0006] Furthermore, DHM undergoes several processing steps, including pooling, pasteurisation or sterilisation and freezing. There is some evidence suggesting that such processing steps may lead to nutritional deficiencies in DHM when compared with MOM, particularly with reference to light and heat sensitive micronutrients. To address the above- mentioned nutritional deficiencies, DHM is often supplemented with a bovine milk-derived or human milk-derived human milk fortifier (HMF), which typically contains protein, carbohydrate, micronutrients (e.g. vitamins, minerals, etc.) and other trace elements.

[0007] Micronutrients and other trace elements play a critical role in growth outcomes of preterm infants, particularly VLBW infants. For example, sodium plays a multi-faceted role in maintenance of intravascular volume, bone mineralisation and nitrogen retention, which all participate in regulating growth. VLBW infants have a limited capacity to regulate urine sodium retention and excretion and this feature is more pronounced in extremely preterm infants when compared to more mature infants. Sodium supplementation after the first week of life allows for adequate diuresis and avoids the potential free water retention that is closely linked to poor respiratory outcomes in this population.

[0008] The inventors of the present application have identified that the micronutrient profile of commercially available DHM obtained from human milk banks does not meet the nutritional needs of preterm infants, even when fortified with commercially available HMFs. A primaryreason for this is the source of the donor milk, which is conventionally from mothers of term infants. Research has demonstrated that the nutrient profile of human milk (HM) is dynamic and is influenced by many factors, including the age of the infant.

[0009] The inventors have identified that the commercially available DHM has significantly lower levels of sodium, chloride, magnesium and zinc than the levels found in early or mature preterm milk. Micronutrients such as sodium and zinc decrease in concentration over the course of lactation. Concentrations of micronutrients in DHM are considerably lower than the concentrations in MOM because donors tend to be at least 6 months post-partum and at a lactation stage in which micronutrient concentrations are at their lowest. In addition, micronutrients tend to be volume dependent, meaning that the more volume of milk that is produced, the lower the concentration of micronutrients per decilitre. Volume of MOM increases as her baby ages, to match the demands of the infant. The volume expressed by a mother at six-months post-partum is considerably more than the milk produced by a mother of a newborn premature infant. Therefore, the micronutrient concentration will be more dilute. In addition, the milk volumes consumed by the preterm infant are considerably lower than the volumes consumed by older, more mature infants. The preterm infant is dependent upon more nutrient dense milk to meet their needs in a limited volume.

[0010] The inventors have identified that milk from mother’s delivering prematurely has three times the concentration of sodium and two times the concentration of chloride, and more than twice the concentration of zinc than does DHM. The inventors concluded that preterm infants whose sole source of nutrition is DHM (often pooled from term mothers), even when fortified with HMF, will likely have a micronutrient deficiency.

[0011] Furthermore, the inventors have determined that commercially available DHM has significantly lower levels of vitamin A, vitamin C, docosahexaenoic acid (DHA), and inositol than the levels found in early or mature preterm milk.

[0012] Vitamin A is essential for eye development, growth, healing and the prevention of bronchopulmonary dysplasia and respiratory infections, conditions that are common in preterm infants.

[0013] The inventors have identified that, whilst the levels of vitamin A in commercially available DHM are consistent with levels in term HM, the levels are insufficient to meet the recommended daily intake (35-1000 mg / 100kcal) for a preterm infant, even following fortification with commercially available HMFs.

[0014] Vitamin C enhances the absorption of non-haem iron, is an important antioxidant and is required for collagen synthesis. The recommended daily intake of vitamin C for a preterm infant is 18-50 mg / 100 kcal.

[0015] The inventors have discovered that vitamin C is undetectable in commercially available DHM and, accordingly, DHM is insufficient to meet the recommended daily intake (18- 50mg / 100kcal) for a preterm infant, even following fortification with commercially available bovine milk-derived HMFs. It should be further noted that human milk-derived HMFs do not contain vitamin C, therefore infants reliant on DHM fortified with human milk-derived HMFs will not receive any vitamin C.

[0016] DHA is a long-chain polyunsaturated fatty acid important for neurodevelopment. Placental transfer of DHA occurs in late pregnancy, meaning preterm infants are at risk of DHA deficiency. DHA is also present in mother’s own milk (MOM) but is highly dependent upon maternal diet.

[0017] The inventors have identified that whilst DHA levels in DHM are comparable to term HM at 4 weeks of lactation, the levels of DHA in >75% DHM samples were below the recommended range (minimum recommended level of 0.5-1 % of total fatty acids) for preterm infants. Hence, even with the fortification with commercially available HMFs, there will be a shortfall in the levels required for the preterm infant, in order to encourage optimal neurodevelopmental outcomes.

[0018] Inositol, primarily the isomer myo-inositol, is a carbocyclic sugar that is abundant in the brain and other mammalian tissues; it mediates cell signal transduction in response to a variety of hormones, neurotransmitters, and growth factors, and participates in osmoregulation. Inositol is also a multifunctional human milk (HM) nutrient with roles in protection from retinopathy of prematurity and neurodevelopment4.

[0019] The inventors determined that levels of inositol in DHM from US milk banks was comparable with unpasteurised term milk across lactation, suggesting inositol isn’t considerably impacted by pasteurisation. Whilst measured levels meet broad preterm nutritional guidelines (4.4-53 mg / 100 kcal), DHM demonstrates lower inositol levels consistent with later-stage lactation (3+ months) HM. This highlights a potential gap in inositol provided by DHM during the critical first months of preterm life when DHM is provided as alternative to MOM.

[0020] As discussed above, commercially available HMFs have been designed to fortify the nutrient content of DHM, but this design has failed to take into account the dynamic nature of HM i.e. the fact that the MOM of a mother of a preterm infant would inherently have a different composition to that of a term mother. HMF fortification for preterm infants often focuses on addressing the shortfall in macronutrients (e.g. protein and fat) and not in micronutrients (e.g. minerals and vitamins).

[0021] If an infant is identified as being deficient in a micronutrient, supplementation generally takes the form of the administration of bolus doses, as a medication. These supplementations come directly from the pharmacy, they are usually compounded, and administered via syringe into a feeding tube.

[0022] Currently no products exist that enable the tailored supplementation of specific micronutrients identified as being deficient in a preterm infant, directly into fortified DHM or MOM (whereby fortification is using standard commercial HMF).

[0023] This has led the inventors to recognise that a need exists for a simple and rapid means of tailoring and further supplementing the fortification of DHM or MOM to address these micronutrient shortfalls, particularly because fortified DHM or MOM is often the sole source of nutrition for a preterm infant, particularly a VLBW infant.Summary of the invention

[0024] The inventors of the present application have identified that the levels of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA, and inositol are significantly lower within DHM than MOM from a mother of a preterm infant. As a result, even when DHM is fortified with commercially available HMF, a clinical deficiency for these micronutrients in the infant is a likely outcome.

[0025] A preterm infant will be regularly monitored for signs of a micronutrient deficiency, and if any symptoms are identified, receive the appropriate micronutrients. However, as the micronutrient supplementations often come directly from the pharmacy, this may result in delay to intervention and also the need for a prescription. Furthermore, the bolus doses of the micronutrients are conventionally administered via syringe into a feeding tube, and as notedabove, preterm infants can only tolerate total daily feedings of between 100 and 150 ml per kg of the infant's weight. Bolus doses of prepared micronutrients from the pharmacy are typically delivered twice per day and may result in intolerance because of their high osmotic load. By diluting the micronutrients into human milk (DHM or MOM) the osmotic load is decreased and the micronutrients can be delivered in smaller, less concentrated doses throughout the day, therefore, reducing the risk of intolerance.

[0026] The inventors have therefore recognised the need for a simple and convenient method to directly supplement MOM, DHM, a MOM / HMF mixture, and / or a DHM / HMF mixture with the appropriate amounts of any deficient micronutrients. This is solved by the HMF micronutrient supplementation kit according to the invention detailed below.

[0027] The HMF micronutrient supplementation kit detailed subsequently provides a tailored approach to micronutrient supplementation based on an individual infant’s requirements.

[0028] According to a first aspect of the invention, there is a human milk fortifier (HMF) micronutrient supplementation kit comprising or consisting of at least two micronutrients, wherein the at least two micronutrients are selected from the group consisting of sodium, chloride, zinc, magnesium, vitamin A, vitamin C, inositol and docosahexaenoic acid (DHA).

[0029] In some embodiments, the HMF micronutrient supplementation kit is for use in supplementing the levels of at least two micronutrients within Mother’s Own Milk (MOM), Donor Human Milk (DHM), a mixture of MOM and HMF (MOM / HMF), and / or within a mixture of DHM and HMF (DHM / HMF).

[0030] In some embodiments, the at least two micronutrients are provided as a blend. Accordingly, the blend of the at least two micronutrients comprises or consists of: a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0031] In a preferred embodiment, the blend of the at least two micronutrients comprises or consists of sodium and chloride.

[0032] In some embodiments, the blend of the at least two micronutrients consists of two micronutrients selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0033] In some embodiments, the blend of the at least two micronutrients consists of three micronutrients selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0034] In some embodiments, the blend of the at least two micronutrients consists of four micronutrients selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0035] In some embodiments, the blend is provided in a powder or a liquid form. In some embodiments, the blend is provided in a dosage unit selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet. In a preferred embodiment, the blend is a powder in a dosage unit. A powder format advantageously enables the micronutrient blend to be dissolved directly into MOM, DHM, a MOM / HMF mixture, and / or a DHM / HMF mixture. Other formats are envisaged, such as a liquid format, which can also be readily incorporated into the MOM, DHM, MOM / HMF mixture, and / or a DHM / HMF mixture.

[0036] In some embodiments, the HMF micronutrient supplementation kit comprises a plurality of dosage units, wherein each dosage unit comprises a blend of the at least two micronutrients. In some embodiments, the HMF micronutrient supplementation kit comprises at least two dosage units, at least five dosage units, at least 10 dosage units, at least 20 dosage units, at least 24 dosage units, at least 25 dosage units, or at least 50 dosage units. In some embodiments, the HMF micronutrient supplementation kit comprises 2 to 100 dosage units, 2 to 50 dosage units, 2 to 25 dosage units, 2 to 24 dosage units, 5 to 100 dosage units, 5 to 50 dosage units, 5 to 25 dosage units, 10 to 100 dosage units, 10 to 50 dosage units, or 10 to 25 dosage units.

[0037] In some embodiments, the at least two micronutrients are provided separately as a discrete dosage units comprising a first dosage unit and a second dosage unit. The first dosage unit comprises a first micronutrient and the second dosage unit comprises a second micronutrient, wherein the first micronutrient is a different micronutrient to that of the second micronutrient. Accordingly, in some embodiments, the HMF micronutrient supplementation kit comprises or consists of: a first dosage unit comprising or consisting of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a second dosage unit comprising or consisting of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0038] In some embodiments, the HMF micronutrient supplementation kit comprises or consists of: a plurality of first dosage units, wherein each of the first dosage units comprises or consists of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a plurality of second dosage units, wherein each of the second dosage units comprises or consists of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0039] In some embodiments, the at least two micronutrients are provided in a powder form, a liquid form, or wherein at least one micronutrient is provided in a powder form and at least one micronutrient is provided in a liquid form. In some embodiments, the at least two micronutrients are provided in a dosage unit selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet. In some embodiments, the first dosage unit may be provided in the same form (e.g. stick pack) as the second dosage unit. Alternatively, the first dosage unit may be provided in a different format to the second dosage unit. A powder format advantageously enables the micronutrient to be dissolved directly into MOM, DHM, a MOM / HMF mixture, and / or a DHM / HMF mixture. Other formats are envisaged, such as a liquid format, which can also be readily incorporated into MOM, DHM, a MOM / HMF mixture, and / or a DHM / HMF mixture. In a preferred embodiment, the at least two micronutrients are both provided as a powder in separate discrete dosage units.

[0040] In some embodiments, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units and second dosage units, wherein the first dosage units each comprise a first micronutrient and the second dosage units each comprise a second micronutrient. In some embodiments, the HMF micronutrient supplementation kit comprises at least two dosage units of each of the first dosage units and second dosage units, at least five dosage units, at least 10 dosage units, at least 20 dosage units, at least 24 dosage units, at least 25 dosage units, or at least 50 dosage units. In some embodiments, the HMF micronutrient supplementation kit comprises 2 to 100 dosage units of each of the first dosage units and the second dosage units, 2 to 50 dosage units, 2 to 25 dosage units, 2 to 24 dosageunits, 5 to 100 dosage units, 5 to 50 dosage units, 5 to 25 dosage units, 10 to 100 dosage units, 10 to 50 dosage units, or 10 to 25 dosage units.

[0041] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, the HMF micronutrient supplementation kit may take the unit of a container, such as a box, container, or tub, in which the above-described dosage units are housed. The HMF micronutrient supplementation kit may be provided with the HMF as an accessory kit or may be ordered as a separate item.

[0042] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise sodium in an amount of 0.1 to 50 mg, 1 to 50 mg, 1 to 40 mg, 1 to 30 mg, 1 to 25 mg, or 5 to 25 mg. In some embodiments, each dosage unit may comprise sodium in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise sodium in the form of sodium chloride, sodium citrate, sodium carbonate, and / or any other suitable form of sodium.

[0043] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise chloride in an amount of 0.1 to 50 mg, 1 to 50 mg, 1 to 40 mg, 1 to 30 mg, 1 to 25 mg, or 5 to 25 mg. In some embodiments, each dosage unit may comprise chloride in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise chloride in the form of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, choline chloride, and / or any other suitable form of chloride.

[0044] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise magnesium in an amount of 0.01 to 5 mg, 0.05 to 5 mg, 0.05 to 2.5 mg, 0.1 to 2.5 mg, 0.1 to 2 mg, or 0.1 to 1.5 mg. In some embodiments, each dosage unit may comprise magnesium in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 1.5 mg, 2.5 mg, or 5 mg. In some embodiments, each dosage unit may comprise magnesium in the form of magnesium chloride, magnesium phosphate dibasic, magnesium oxide, and / or any other suitable form of magnesium.

[0045] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise zinc in an amount of 0.01 to 5 mg, 0.01 to 2.5 mg, 0.01 to 2 mg, 0.01 to 1 mg, 0.05 to 1 mg, 0.1 to 1 mg, 0.1 to 0.75 mg, or 0.1 to 0.5 mg. In some embodiments, each dosage unit may comprise zinc in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.75 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg or 5 mg. In some embodiments, each dosage unit may comprise zinc in the form of zinc sulphate, zinc sulphate monohydrate, and / or any other suitable form of zinc.

[0046] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise vitamin A in an amount of 0.01 to 5 mg, 0.01 to 2.5 mg, 0.01 to 2 mg, 0.01 to 1 mg, 0.01 to 0.5 mg, 0.01 to 0.25 mg, or 0.05 to 0.25 mg. In some embodiments, each dosage unit may comprise vitamin A in an amount of 0.01 mg, 0.05 mg, 0.075 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.5 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg or 5 mg. In some embodiments, each dosage unit may comprise vitamin A in the form of vitamin A palmitate (liquid and / or beadlet), beta-carotene, and / or any other suitable form of vitamin A.

[0047] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise vitamin C in an amount of 0.1 to 50 mg, 0.1 to 40 mg, 0.1 to 30 mg, 0.1 to 25 mg, 1 to 25 mg, 1 to 20 mg,or 1 to 15 mg. In some embodiments, each dosage unit may comprise vitamin C in an amount of 0.1 mg, 0.5 mg, 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise vitamin C in the form of ascorbic acid, sodium ascorbate, and / or any other suitable form of vitamin C.

[0048] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise DHA in an amount of 0.1 to 50 mg, 0.1 to 40 mg, 0.1 to 30 mg, 0.1 to 25 mg, 0.5 to 25 mg, 0.5 to 20 mg, or 0.5 to 15 mg. In some embodiments, each dosage unit may comprise DHA in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 8 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg.

[0049] In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise inositol in an amount of 0.01 to 50 mg, 0.1 to 50 mg, 0.1 to 40 mg, 0.1 to 30 mg, 0.1 to 25 mg, 0.1 to 20 mg, 0.1 to 10 mg, or 0.5 to 10 mg. In some embodiments, each dosage unit may comprise inositol in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 7.5 mg, 10 mg, 20 mg, 25 mg, or 50 mg.

[0050] In a preferred embodiment, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units each comprising or consisting of sodium and a plurality of second dosage units each comprising or consisting of chloride.

[0051] In a preferred embodiment, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units each comprising or consisting of zinc, a plurality of second dosage units each comprising or consisting of vitamin C.

[0052] In a hospital, it is common practice to calculate preparations of human milk (MOM and / or DHM) based on increments of 30mL. When preparing the patient's feed, it is typical practice to add HMF and / or additional macro / micronutrients to the human milk to compensate for any macro / micronutrient deficiencies and thus, meet expert nutrient level recommendations for preterm infants. Therefore, in some embodiments, each dosage unit contains the desired amount of the particular micronutrient or micronutrient blend for a single 30mL increment of human milk, which may or may not contain HMF. Alternatively, in some embodiments, each dosage unit contains the desired amount of the particular micronutrient or micronutrient blend for multiple 30mL increments of human milk, which may or may not contain HMF. Accordingly, each dosage unit may comprise at least two 30mL increments of the particular micronutrient or micronutrient blend, at least three increments, at least four increments, at least five increments, at least 10 increments, at least 20 increments, at least 25 increments, or at least 50 increments. Accordingly, each dosage unit may comprise 2 to 50 30mL increments of the particular micronutrient or micronutrient blend, 2 to 25 increments, 2 to 20 increments, 2 to 10 increments, or 2 to 5 increments.

[0053] In some preferred embodiments, the HMF micronutrient supplementation kit comprises or consists of at least three micronutrients, wherein each of the at least three micronutrients are different micronutrients. Therefore, the HMF micronutrient supplementation kit comprises or consists of: a first dosage unit, wherein said first dosage unit comprises or consists of a first micronutrient; a second dosage unit, wherein said second dosage unit comprises or consists of a second micronutrient; and a third dosage unit, wherein said third dosage unit comprises or consists of a third micronutrient.

[0054] In some preferred embodiments, the HMF micronutrient supplementation kit comprises or consists of at least three micronutrients, wherein each of the at least three micronutrients are different micronutrients. Therefore, the HMF micronutrient supplementation kit comprises or consists of: a plurality of first dosage units, wherein each of the first dosage units comprise or consist of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; a plurality of second dosage units, wherein each of the second dosage units comprise or consist of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; and a plurality of third dosage units, wherein each of the third dosage units comprise or consist of a third micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0055] In a preferred embodiment, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units each comprising or consisting of sodium, a plurality of second dosage units each comprising or consisting of zinc, and a plurality of third dosage units each comprising or consisting of vitamin C.

[0056] In some preferred embodiments, the HMF micronutrient supplementation kit comprises or consists of at least four micronutrients, wherein each of the at least four micronutrients are different micronutrients. Therefore, the HMF micronutrient supplementation kit comprises or consists of: a first dosage unit, wherein said first dosage unit comprises or consists of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; a second dosage unit, wherein said second dosage unit comprises or consists of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; a third dosage unit, wherein said third dosage unit comprises or consists of a third micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; and a fourth dosage units, wherein said fourth dosage unit comprises or consists of a fourth micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0057] In some preferred embodiments, the HMF micronutrient supplementation kit comprises or consists of at least four micronutrients, wherein each of the at least four micronutrients are different micronutrients. Therefore, the HMF micronutrient supplementation kit comprises or consists of: a plurality of first dosage units, wherein each of the first dosage units comprise or consist of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; a plurality of second dosage units, wherein each of the second dosage units comprise or consist of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; a plurality of third dosage units, wherein each of the third dosage units comprise or consist of a third micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol; and a plurality of fourth dosage units, wherein each of the fourth dosage units comprise or consist of a fourth micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0058] In a preferred embodiment, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units each comprising or consisting of sodium, a pluralityof second dosage units each comprising or consisting of zinc, a plurality of third dosage units each comprising or consisting of vitamin C, and a plurality of fourth dosage units each comprising or consisting of chloride.

[0059] In some preferred embodiments, the HMF micronutrient supplementation kit comprises or consists of a plurality of first dosage units each comprising or consisting of sodium, a plurality of second dosage units each comprising or consisting of chloride, a plurality of third dosage units each comprising or consisting of zinc, and a plurality of fourth dosage units each comprising or consisting of magnesium.

[0060] In some embodiments, in addition to the at least two micronutrients selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, a further micronutrient is added, such as potassium. In some embodiments, including wherein the at least two micronutrients are provided as a blend or as separate discrete dosage units, each dosage unit may comprise potassium in an amount of 0.1 to 50 mg, 1 to 50 mg, 1 to 40 mg, 1 to 30 mg, 1 to 25 mg, or 1 to 20 mg. In some embodiments, each dosage unit may comprise potassium in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise potassium in the form of potassium chloride, potassium citrate, potassium bicarbonate, potassium phosphate, and / or any other suitable form of potassium.

[0061] A preterm infant will be regularly monitored for signs of a vitamin deficiency, and if any symptoms are identified, receive the appropriate vitamins.

[0062] According to a second aspect of the invention, there is provided a nutritional composition for administration to an infant, wherein the composition comprises or consists of:(a) human milk fortifier; and(b) a first micronutrient and a second micronutrient, wherein each of the first and second micronutrients are added to the composition as a micronutrient supplementation.

[0063] The nutritional composition may be administered to a preterm infant, in particular to a VLBW infant.

[0064] The composition may comprise human milk. The human milk may be MOM or DHM.

[0065] The at least two micronutrients are provided by the HMF micronutrient supplementation kit of the first aspect of the invention.

[0066] According to a third aspect of the invention, there is provided a method of supporting the growth of an infant by administering a composition comprising or consisting of:(a) human milk fortifier; and(b) a micronutrient supplementation comprising or consisting of at least two micronutrients, wherein the infant is deficient in the at least two micronutrients.

[0067] In some embodiments of the method, the infant is a preterm infant, in particular a VLBW infant.

[0068] In some embodiments of the method, the composition comprises human milk. In some embodiments of the method, the human milk is MOM or DHM.

[0069] The at least two micronutrients are provided by the HMF micronutrient supplementation kit of the first aspect of the invention.

[0070] An HCP when noticing signs / symptoms of a micronutrient deficiency in an infant, or when noticing deficient levels of a micronutrient during a routine screen, will be able to readily select the appropriate micronutrient or micronutrient blend from the HMF micronutrient supplementation kit and add this directly to the MOM / HMF or DHM / HMF. The HCP may become aware of a micronutrient deficiency by the results of an electrolyte blood test. The micronutrient deficiency may be noted during a routine screen by way of the result of a blood and / or urine electrolyte test. Accordingly, in some embodiments of the invention, the method may therefore include the step of measuring electrolytes in the blood of an infant.

[0071] In some embodiments, the method is a non-therapeutic method, supporting normal growth rather than treating a clinical condition relating to impaired growth.

[0072] In some embodiments of the method, a HMF may be provided with a HMF micronutrient supplementation kit that includes at least 10 stick packs, with each stick pack including a powder micronutrient blend of, for example 1 mg sodium salt and 1 mg zinc salt. The sodium salt may be sodium chloride. The zinc salt may be zinc sulphate. The powder from one or more stick packs can be dissolved directly into the MOM, DHM, MOM / HMF, and / or DHM / HMF if the infant requires supplementation with sodium, chloride and zinc. It is envisaged that each stick pack may include higher amounts of the micronutrients, non-limiting examples include 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg or 10 mg. It is also envisaged that each stick pack may also include micronutrients in multiples of 5 mg, for example 5 mg, 10 mg, 15 mg and 20 mg. It is also envisaged that each stick pack may also include micronutrients in multiples 10mg, for example 10 mg, 20 mg, 30 mg, 40 mg and 50 mg.

[0073] In an alternative embodiment, the HMF may be provided with a HMF micronutrient supplementation kit that includes at least 10 stick packs of 1 mg sodium salt and at least 10 stick packs of 1 mg zinc salt. The sodium salt may be sodium chloride. The zinc salt may be zinc sulphate. If the infant only requires 1 mg sodium supplementation, the HCP can dissolve the contents of 1 stick pack of sodium salt into the MOM, DHM, MOM / HMF, and / or DHM / HMF. If the infant requires 2 mg sodium supplementation, the HCP can dissolve the contents of 2 stick packs of sodium salt into the MOM, DHM, MOM / HMF, and / or DHM / HMF. If the infant requires 1mg sodium supplementation and 1 mg of zinc supplementation, the HCP can dissolve the contents of 1 stick pack of sodium, and 1 stick pack of zinc into the MOM, DHM, MOM / HMF, and / or DHM / HMF.

[0074] In an alternative embodiment, the HMF may be provided with a HMF micronutrient supplementation kit that includes: 10 stick packs x 1 mg vitamin A and 10 stick packs x 10 mg vitamin C. If the infant only requires 1 mg vitamin A supplementation, the HCP can dissolve the contents of 1 stick pack of vitamin into the MOM / HMF or DHM / HMF. If the infant requires 2 mg vitamin A supplementation, the HCP can dissolve the contents of 2 stick packs of vitamin A into the MOM / HMF or DHM / HMF. If the infant requires 1 mg vitamin A supplementation and 10 mg of vitamin C supplementation, the HCP can dissolve the contents of 1 stick pack of vitamin A, and 1 stick pack of vitamin C into the MOM / HMF or DHM / HMF.

[0075] According to a fourth aspect of the invention, there is provided a human milk fortifier (HMF) micronutrient supplementation kit comprising or consisting of a single micronutrient, wherein the HMF micronutrient supplementation kit is for use in supplementing the levels of the micronutrient within Mother’s Own Milk (MOM), Donor Human Milk (DHM), a mixture of MOM and HMF (MOM / HMF, and / or within a mixture of DHM and HMF (DHM / HMF).

[0076] The HMF micronutrient supplementation kit according to the fourth aspect, wherein the single micronutrient is selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA.

[0077] In some embodiments, the single micronutrient is provided as a discrete dosage unit. In some embodiments, the HMF micronutrient supplementation kit comprises a plurality of dosage units wherein each of the dosage units comprises or consists of a micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

[0078] In some embodiments, the dosage unit comprises a first dosage unit and a second dosage unit. The first and second dosage units comprise the same micronutrient but in different dosage amounts. For example, the first dosage unit may comprise or consist of about 1 mg of a micronutrient and the second dosage unit may comprise or consist of about 5 mg unit of the same micronutrient. Accordingly, in some embodiments, the HMF micronutrient supplementation kit according to the fourth aspect comprises or consists of: a first dosage unit comprising or consisting of a micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a second dosage unit comprising or consisting of the same micronutrient as the first dosage unit.

[0079] In some embodiments, the HMF micronutrient supplementation kit of the fourth aspect comprises or consists of: a plurality of first dosage units, wherein each of the first dosage units comprises or consists of a micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a plurality of second dosage units, wherein each of the second dosage units comprises or consists of the same micronutrient as each of the first dosage units.

[0080] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise sodium in an amount of 0.1 to 50 mg, 1 to 50 mg, 1 to 40 mg, 1 to 30 mg, 1 to 25 mg, or 5 to 25 mg. In some embodiments, each dosage unit may comprise sodium in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise sodium in the form of sodium chloride, sodium citrate, sodium carbonate, and / or any other suitable form of sodium. In a preferred embodiment, the HMF micronutrient supplementation kit comprises sodium in the form of sodium chloride.

[0081] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise chloride in an amount of 0.1 to 50 mg, 1 to 50 mg, 1 to 40 mg, 1 to 30 mg, 1 to 25 mg, or 5 to 25 mg. In some embodiments, each dosage unit may comprise chloride in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise chloride in the form of sodium chloride, potassium chloride, calcium chloride, magnesium chloride, choline chloride, and / or any other suitable form of chloride.

[0082] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise magnesium in an amount of 0.01 to 5 mg, 0.05 to 5 mg, 0.05 to 2.5 mg, 0.1 to 2.5 mg, 0.1 to 2 mg, or 0.1 to 1.5 mg. In some embodiments, each dosage unit may comprise magnesium in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.25 mg, 0.5 mg, 0.75 mg, 1 mg, 1.5 mg, 2.5 mg, or 5 mg. In some embodiments, each dosage unit may comprise magnesium in the form of magnesium chloride, magnesium phosphate dibasic, magnesium oxide, and / or any other suitable form of magnesium.

[0083] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise zinc in an amount of 0.01 to 5 mg, 0.01 to 2.5 mg, 0.01 to 2 mg, 0.01 to 1 mg, 0.05 to 1 mg, 0.1 to 1 mg, 0.1 to 0.75 mg, or 0.1 to 0.5 mg. In some embodiments, each dosage unit may comprise zinc in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.2 mg, 0.25 mg, 0.3 mg, 0.4 mg, 0.5 mg, 0.75 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg or 5 mg. In some embodiments, each dosage unit may comprise zinc in the form of zinc sulphate, zinc sulphate monohydrate, and / or any other suitable form of zinc.

[0084] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise vitamin A in an amount of 0.01 to 5 mg, 0.01 to 2.5 mg, 0.01 to 2 mg, 0.01 to 1 mg, 0.01 to 0.5 mg, 0.01 to 0.25 mg, or 0.05 to 0.25 mg. In some embodiments, each dosage unit may comprise vitamin A in an amount of 0.01 mg, 0.05 mg, 0.075 mg, 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.5 mg, 1 mg, 1.5 mg, 2 mg, 2.5 mg or 5 mg. In some embodiments, each dosage unit may comprise vitamin A in the form of vitamin A palmitate (liquid and / or beadlet), beta-carotene, and / or any other suitable form of vitamin A.

[0085] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise vitamin C in an amount of 0.1 to 50 mg, 0.1 to 40 mg, 0.1 to 30 mg, 0.1 to 25 mg, 1 to 25 mg, 1 to 20 mg, or 1 to 15 mg. In some embodiments, each dosage unit may comprise vitamin C in an amount of 0.1 mg, 0.5 mg, 1 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg. In some embodiments, each dosage unit may comprise vitamin C in the form of ascorbic acid, sodium ascorbate, and / or any other suitable form of vitamin C.

[0086] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise DHA in an amount of 0.1 to 50 mg, 0.1 to 40 mg, 0.1 to 30 mg, 0.1 to 25 mg, 0.5 to 25 mg, 0.5 to 20 mg, or 0.5 to 15 mg. In some embodiments, each dosage unit may comprise DHA in an amount of 0.1 mg, 0.5 mg, 1 mg, 2 mg, 2.5 mg, 5 mg, 7.5 mg, 8 mg, 10 mg, 15 mg, 20 mg, 25 mg, or 50 mg.

[0087] In some embodiments, including wherein the single micronutrient is provided as discrete dosage units, each dosage unit may of the HMF micronutrient supplementation kit of the fourth aspect may comprise inositol in an amount of 0.01 to 50 mg, 0.01 to 40 mg, 0.01 to 30 mg, 0.01 to 25 mg, 0.01 to 20 mg, 0.01 to 10 mg, or 0.05 to 10 mg. In some embodiments, each dosage unit may comprise inositol in an amount of 0.01 mg, 0.05 mg, 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 7.5 mg, 10 mg, 20 mg, 25 mg, or 50 mg.

[0088] In some embodiments, the single micronutrient is provided in a powder or a liquid form. In some embodiments, the single micronutrient is provided in a dosage unit selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet. In a preferred embodiment, the single micronutrient is a powder in a dosage unit. A powder format advantageously enables the micronutrient blend to be dissolved directly into MOM, DHM, a MOM / HMF mixture, and / ora DHM / HMF mixture. Otherformats are envisaged, such as a liquid format, which can also be readily incorporated into the MOM, DHM, MOM / HMF mixture, and / or a DHM / HMF mixture.

[0089] In some embodiments, the HMF micronutrient supplementation kit of the fourth aspect comprises a plurality of dosage units, wherein each dosage unit comprises the single micronutrient. In some embodiments, the HMF micronutrient supplementation kit comprises at least two dosage units, at least five dosage units, at least 10 dosage units, at least 20 dosage units, at least 24 dosage units, at least 25 dosage units, or at least 50 dosage units.In some embodiments, the HMF micronutrient supplementation kit comprising a single micronutrient comprises 1 to 100 dosage units, 1 to 50 dosage units, 1 to 25 dosage units, 1 to 24 dosage units, 5 to 100 dosage units, 5 to 50 dosage units, 5 to 25 dosage units, 10 to 100 dosage units, 10 to 50 dosage units, or 10 to 25 dosage units.Brief Description of the Drawings

[0090] The accompanying Figures, which are incorporated into and constitute part of this specification, are illustrative of the invention.

[0091] Figure 1 : Comparison of sodium (Na) content in in early (D7, n = 36, blue circles) and mature (D28, n = 28, red circles) preterm human milk and in donor human milk (DHM, n = 15, green squares). Black bar represents mean for each group. For all graphs, *P < 465 0.05, **P< 0.01 , ***P < 0.001 , ****p < 0.0001. Analysis by 1-way ANOVA with Dunnett’s 466 correction for multiple comparisons.

[0092] Figure 2: Comparison of chloride (Cl) content in early (D7, n = 36, blue circles) and mature (D28, n = 28, red circles) preterm human milk and in donor human milk (DHM, n = 15, green squares). Black bar represents mean for each group. For all graphs, *P < 465 0.05, **P< 0.01 , ***P < 0.001 , ****p < 0.0001. Analysis by 1-way ANOVA with Dunnett’s 466 correction for multiple comparisons.

[0093] Figure 3: Comparison of magnesium (Mg) content in early (D7, n = 36, blue circles) and mature (D28, n = 28, red circles) preterm human milk and in donor human milk (DHM, n = 15, green squares). Black bar represents mean for each group. For all graphs, *P < 465 0.05, **P < 0.01 , ***p < 0.001 , 0.0001. Analysis by 1 -way ANOVA with Dunnett’s 466 correction for multiple comparisons.

[0094] Figure 4: Comparison of zinc (Zn) content in early (D7, n = 36, blue circles) and mature (D28, n = 28, red circles) preterm human milk and in donor human milk (DHM, n = 15, green squares). Black bar represents mean for each group. For all graphs, *P < 4650.05, **P < 0.01 , ***P < 0.001 , ****p < 0.0001. Analysis by 1-way ANOVA with Dunnett’s 466 correction for multiple comparisons.

[0095] Figure 5: Comparison of inositol content in donor human milk (n=15) with unpasteurised term human milk at five distinct lactation stages (n=30 per each week of lactation). Density curves for DHM and each time point represent sample-to-sample variability. The median for DHM is represented by the vertical dashed line with 95% quantiles shown by dotted lines. DHM is most consistent with unpasteurised term milk from 13-26 weeks lactation (approximately 3-6 months).Detailed Description of The Preferred Embodiments

[0096] Reference will now be made in detail to embodiments of the invention, one or more examples are set forth below. Each example is provided by way of explanation of the invention, not limitation of the invention. In fact, it will be apparent to those in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. For instance, features illustrated or described as part of one embodiment can be used on another embodiment to yield a still further embodiment.

[0097] As used herein the term “infant’ means a human subject ranging in age from birth to not more than one year and includes infants from 0 to 12 months corrected age. The phrase “corrected age" means an infant’s chronological age minus the amount of time that the infant was born premature. Therefore, the corrected age is the age of the infant if it had been carried to full term. The term infant includes full-term infants, preterm infants, low birth weight infants,very low birth weight infants, and extremely low birth weight infants. “Preterm" means an infant born before the end of the 37thweek of gestation. “Full-term" means an infant born after the end of the 37thweek of gestation.

[0098] The term “human milk fortifier1’, also known as “human milk supplementation", refers to an additive for human milk. In some embodiments, the human milk fortifier or supplementation may improve the nutritional content of certain components of human milk.

[0099] The term “powder" as used herein refers to dry particles produced by the grinding, crushing, or disintegration of a solid substance. The particles may also be referred to as granules or particulates.

[0100] All references to singular characteristics or limitations in the present disclosure shall include the corresponding plural characteristic or limitation, and vice versa, unless otherwise specified or clearly implied to the contrary, by the context in which the reference is made.

[0101] The compositions of the present disclosure and the HMF micronutrient supplementation kit can comprise, consist of, or consist essentially of any of the components, ingredients, micronutrients, dosage units, etc. described herein, as well as including any additional useful component, ingredient, micronutrient, dosage unit, etc.EXAMPLE 1 : COMPARISON OF MACRO- AND MICRONUTRIENT PROFILE OF COMMERCIAL DHM WITH EARLY AND MATURE PRE-TERM MILKMETHODOLOGY(a) Commercial DHM

[0102] Commercial DHM was purchased from Ni-Q (Portland, OR), Prolacta Bioscience (Duarte, CA), and the Human Milk Banking Association of North America (HMBANA; five locations provided 13 samples). Ni-Q HDM+ is a sterile milk product that is stored at room temperature while Prolacta HM undergoes “high temperature, short time” pasteurisation and HMBANA uses Holder Pasteurisation for all pooled DHM. All DHM was stored at -20°C (except shelf stable Ni-Q) and were shipped frozen overnight to Eurofins S-F Laboratories (New Berlin, Wl) for analysis. All sample analysis met the standards outlined by the Official Methods of Analysis (AOAC). The study was reviewed by the Augusta University Institutional Review Board and was registered with ClinicalTrials.gov (NCT05742815).(b) Preterm Human Milk Samples

[0103] Raw data from preterm human milk samples collected during a previous study were used for comparisons with DHM. The study was approved by the Augusta University Institutional Review Board (IRB). Eligible mothers were identified as women who delivered preterm (< 33 weeks), whose infant was admitted to the NICU at the Children’s Hospital of Georgia prior to day of life 7 and intended to provide expressed human milk to their infant. Written united consent was obtained from all mothers and assent was requested prior to each study sample to ensure continued willingness to participate and provide samples. Mothers were excluded from the study if 1) removing the study volume of (20mL) of expressed breastmilk one time per week contributed to inadequate milk supply to meet the nutritional needs of the infant, 2) the mother identified as vegan or vegetarian, 3) she restricted her daily caloric intake to <1200 calories per day, or 4) she was positive for the Human Immunodeficiency Virus (HIV). Expressed milk samples were collected on post-partum days 7, 14, 21 , and 28 between January and November 2019. Samples from post- partum day 7 (n = 36) were defined as “early” and post-partum day 28 (n = 28) as “mature” preterm human milk. Please see flow chart for details. Samples were collected in polypropylene bottles andmothers were instructed to fully empty both breasts every 3 hours. Each 24-hour collection was pooled in a single-use container and a 20-mL sample was extracted from the 24-hour pooled 76 sample for analysis.(c) Ion Content

[0104] Samples were analysed for magnesium (Mg), sodium (Na) and zinc (Zn) by ICP-OES in accordance with EPA method SW-846 601 OB (1996). Briefly, a portion of each sample was microwave digested with nitric acid and hydrochloride acid. Digested samples were analysed for the above metals and micronutrients by Inductively Coupled Plasma-Optical Emission Spectroscopy (ICP-OES). Chloride (Cl) analysis was analysed according to AOAC official method 963.05 and 986.26. Samples were acidified with nitric acid. Chloride was determined potentiometrically by either manual or auto titration with a silver nitrate solution.(d) Statistical Analysis

[0105] Raw data from early (day 7) and mature (day 28) preterm milk were compared to DHM data using one-way ANOVA with Dunnett’s correction for multiple comparisons using PRISM 9.5. Statistical significance was set at P < 0.05.RESULTS

[0106] FIGS. 1-4 illustrate that sodium (FIG. 1), chloride (FIG. 2), and zinc (FIG. 4) levels are reduced in DHM when compared with either early or mature preterm milk, whilst magnesium content was significantly higher in mature preterm milk but did not differ between early preterm milk and DHM (see, FIG. 3).

[0107] Transition to the extrauterine environment is associated with major changes of body water and salt composition in the premature baby.

[0108] The relatively low sodium content in DHM undergirds the slow growth observed in VLBW infants provided an exclusive DHM diet. Preterm human milk contains much higher sodium concentrations than observed in term human milk and sodium content appears to diminish in term human milk over time.

[0109] A recent meta-analysis conducted by the National Academy of Sciences2estimated mean sodium content in term HM is estimated to be 13.7 mg / dL (95% Cl: 11.8 - 15.6 mg / dL) during the first 6 months after birth and 11.1 mg / dL (95% Cl: 7.8 - 14.3 mg / dL) in later term human milk. The later values are most consistent with the data shown in FIG. 1 , reinforcing that commercially available DHM is conventionally derived from pools of mature term milk.

[0110] Multi-nutrient HMFs are added to HM to offset potential nutrient deficits resulting from an exclusive HM diet. However, multi-nutrient HMFs are designed to augment the composition of preterm mother’s own milk, which contains twice as much sodium as DHM. Thus, HMF- fortified DHM may not meet the daily sodium requirements for preterm infants.

[0111] Chloride (Cl’) is the major anion of the extracellular fluids. In preterm infants, chloride balance is influenced by gestational age and by interrelationship between sodium and chloride intake. Concentrations of chloride are significantly lower in DHM (30.9 mg / dL) compared with preterm MOM (69.5 mg / dL). Inadequate intake of chloride during infancy is associated with metabolic acidosis and unfavourable neonatal outcomes3.

[0112] Based on limited clinical data, enteral zinc intake of at least 2 mg / kg / day is recommended with some extremely preterm infants requiring as much as 3 mg / kg / day due to faster growth velocity. Zinc appears to play a particularly important role in linear growth with arecent meta-analysis suggesting that zinc supplementation increases length by nearly 2 cm in low-birth-weight infants (< 2,500 grams) by term-corrected age. Even more compelling, the cumulative mortality rate was significantly lower in the zinc-treatment group when compared with controls (RR 0.55, 95% Cl: 0.31 - 0.97). Although DHM contains very low quantities of zinc, preterm human milk does not contain sufficient zinc concentrations to reach the recommended minimum enteral intake of 2 mg / kg / day.EXAMPLE 2: ANALYSIS OF VITAMIN A IN DHMMETHODOLOGY(a) Commercial DHM

[0113] Commercial DHM was purchased from two for-profit milk banks (NiA, Prolacta Bioscience) and five not-for-profit milk banks (Human Milk Banking Association of North America) representing seven states across the US.(b) Analysis

[0114] Fifteen DHM samples from seven human milk banks were analysed. Samples were shipped to Eurofins SF Analytical Laboratories (New Berlin, Wl) and analysed for vitamin A concentration by AOAC 992.04, 992.06 and 2001.13 methodology using UHPLC.RESULTS

[0115] A mean vitamin A concentration of 119.6 ± 29.5 lU / dL measured within the DHM samples is consistent with reported concentrations in fresh term MOM milk.

[0116] HMFs are typically based on fortifying term HM. The inventors have identified that further supplementation of vitamin A levels will be required to meet the recommended daily intake of 365-1000 mg / 100 kcal of vitamin A for preterm infants whose sole source of nutrition is DHM / HMF and / or MOM / HMF.EXAMPLE 3: ANALYSIS OF VITAMIN C IN DHMMETHODOLOGY(a) Commercial DHM

[0117] Commercial DHM was purchased from two for-profit milk banks (NiA, Prolacta Bioscience) and five not-for-profit milk banks (Human Milk Banking Association of North America) representing seven states across the US.(b) Analysis

[0118] Fifteen DHM samples from seven human milk banks were analysed. Samples were shipped to Eurofins SF Analytical Laboratories (New Berlin, Wl) and were analysed for vitamin C concentration by AOAC 2012.22 methodology using UHPLC.RESULTS

[0119] Vitamin C levels in all fifteen DHM samples were below the level of detection (i.e. <2.5 mg / dL).

[0120] As vitamin C was not detectable in DHM, concentrations will be insufficient for preterm nutrition. Therefore, preterm infants receiving DHM require a supplemental source to provide 100% of their recommended daily intake of 18-50 mg / 100 kcal of vitamin C.

[0121] HMFs are typically based on fortifying term HM. Whilst bovine-milk derived HMF, such as Enfamil HMF (Mead Johnson), includes an amount of vitamin C, the inventors have identified that further supplementation of the vitamin C level in these HMFs will be required for preterm infants. Additionally, the inventors have identified that human-milk derived HMF, such as Prolacta (Prolacta Bioscience), does not contain any vitamin C, and therefore the only source of vitamin C will be by the supplementation of vitamin C from the HMF micronutrient supplementation kit.EXAMPLE 4: ANALYSIS OF DOCOSAHEXAENOIC ACID (DHA) IN DHMMETHODOLOGY(a) Commercial DHM

[0122] Commercial DHM was purchased from two for-profit milk banks (NiA, Prolacta Bioscience) and five not-for-profit milk banks (Human Milk Banking Association of North America) representing seven states across the US.(b) Analysis

[0123] Fifteen DHM samples from seven human milk banks were analysed. Samples were shipped to Eurofins SF Analytical Laboratories (New Berlin, Wl) and were analysed for via gas chromatography using AOAC 996.06 and AOCS Ce official methods.

[0124] Levels were compared to DHA levels from unpasteurised, term HM (n=357) collected at 4 weeks’ lactation from the GEHM study. DHA was extracted from GEHM samples using a modified Bligh-Dyer and analysed via gas chromatography by Cincinnati Children’s Hospital Medical Center.RESULTS

[0125] Two (2) of fifteen DHM samples fell below the DHA quantification limit (i.e. <0.05mg / g).

[0126] As shown in Table 1 , DHA levels in the remaining thirteen DHM samples (DHA mg / dL: mean = 8.76, SD = 3.71) were similar to term HM samples (DHA mg / dL: mean = 8.72, SD = 8.98). However, DHA in ten of thirteen DHM samples was below the minimum recommendation for preterm infants (0.5 — 1 % of total fatty acids).Table 1 : DHA values for DHM and GEHM samples, expressed as mg / dL and as percentage of total fatty acidsEXAMPLE 5: ANALYSIS OF INOSITOL IN DHM(a) Commercial DHM

[0127] Commercial DHM was purchased from two for-profit milk banks (NiA, Prolacta Bioscience) and five not-for-profit milk banks (Human Milk Banking Association of North America) representing seven states across the US.(b) Analysis

[0128] Fifteen DHM samples from seven human milk banks were analysed. Samples were shipped to Eurofins SF Analytical Laboratories (New Berlin, Wl) and were analysed for free myo-inositol via HPAEC-PAD (AOAC 2012.12)

[0129] Levels of inositol in DHM was compared to levels in unpasteurised, term milk (n=150) collected from 10 mothers from each of three populations (Mexico, China, U.S.) over lactation (2-52 weeks) from the GEHM study.RESULTS

[0130] Mean inositol measured in DHM was 13.7 mg / dL and ranged 8.8-18.6 mg / dL. These levels are consistent with mean 13.5 (4.3-25.8) mg / dL inositol for term HM across lactation stages. However, with significant decrease in inositol level in MOM over lactation stage, the inositol level in DHM is most consistent with the level in 13-wk term HM (see FIG. 5). DHM inositol is lower than early-lactation MOM with 24% less inositol than mean 2-wk milk (18.0 mg / dL). Eight of fifteen DHM samples had lower inositol level than mean 3-month MOM and two had a lower inositol level than 6-month milk, pointing to donor lactation stage impacts.

[0131] Non-limiting examples of HMF micronutrient supplementation kits according to the first aspect of the invention are provided below in which a container (e.g. a tub or a box) contains a plurality of stick packs.Kit 1 :Kit 12:Kit 22:Kit 32:Kit 42:Kit 52:Kit 62:Kit 72:Kit 82:Kit 92:Kit 102:References1. Osterman M, Hamilton B, Martin JA, Driscoll AK, Valenzuela CP. Births: Final Data for 306 2020. Natl. Vital Stat. Rep. 2021 ;70(17):1-50.2. Edition ed. In: Oria M, Harrison M, Stallings VA, eds. Dietary Reference Intakes for Sodium and Magnesium. Washington (DC), 2019.3. Christoph Fusch. Water, Sodium, Potassium, and Chloride. Koletzko B, Cheah F-C, Domelldf M, Poindexter BB, Vain N, van Goudoever JB (eds): Nutritional Care of Preterm Infants. Scientific Basis and Practical Guidelines. World Rev. Nutr. Diet. Basel, Karger, 2021 , vol 122, pp 103-121.4. Paquette. A.F, et al. The human milk component myo-inositol promotes neuronal connectivity. PNAS, 2023 vol 120 (30).

Claims

Claims1 . A human milk fortifier (HMF) micronutrient supplementation kit comprising at least two micronutrients, wherein the at least two micronutrients are selected from the group consisting of sodium, chloride, zinc, magnesium, vitamin A, vitamin C, inositol and docosahexaenoic acid (DHA).

2. The HMF micronutrient supplementation kit of claim 1 , wherein the HMF micronutrient supplementation kit is for use in supplementing the levels of the at least two micronutrients within Mother’s Own Milk (MOM), Donor Human Milk (DHM), a mixture of MOM and HMF (MOM / HMF), and / or within a mixture of DHM and HMF (DHM / HMF).

3. The HMF micronutrient supplementation kit of claim 1 or 2, wherein the at least two micronutrients are provided as a blend in a dosage unit.

4. The HMF micronutrient supplementation kit of claim 3, wherein the blend is provided in the form of a powder or a liquid.

5. The HMF micronutrient supplementation kit of claim 3 or 4, wherein the dosage unit is selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet.

6. The HMF micronutrient supplementation kit of any of claims 3 to 5, wherein the HMF micronutrient supplementation kit comprises a plurality of dosage units, preferably 2 to 100 dosage units.

7. The HMF micronutrient supplementation kit of claim 1 or 2, wherein each of the at least two micronutrients is provided separately as discrete dosage units, and wherein the HMF micronutrient supplementation kit comprises: a first dosage unit, wherein said first dosage unit comprises a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA, and a second dosage unit, wherein said second dosage unit comprises a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA.

8. The HMF micronutrient supplementation kit of claim 7, wherein the at least two micronutrients are provided in the form of a powder, a liquid, or at least one micronutrient is provided in the form of a powder and at least one micronutrient is provided in the form of a liquid.

9. The HMF micronutrient supplementation kit of claim 7 or 8, wherein the first dosage unit and / or the second dosage unit is selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet.

10. The HMF micronutrient supplementation kit of any of claims 7 to 9, wherein the first dosage unit comprises a plurality of first dosage units and the second dosage unit comprises a plurality of second dosage units, preferably the HMF micronutrient supplementation kit comprises 2 to 100 first dosage units and 2 to 100 second dosage units.

11. The HMF micronutrient supplementation kit of any of claims 1 to 10, wherein the at least two micronutrients comprises sodium in an amount of 0.1 to 50 mg, chloride in an amount of 0.1 to 50 mg, zinc in an amount of 0.01 to 5 mg, magnesium in an amountof 0.01 to 5 mg, vitamin A in an amount of 0.01 to 5 mg, vitamin C in an amount of 0.1 to 50 mg, DHA in an amount of 0.1 to 50 mg, and / or inositol in an amount of 0.01 to 50 mg.

12. An infant nutritional composition, wherein the composition comprises:(a) human milk fortifier; and(b) at least two micronutrients, wherein the at least two micronutrients comprises a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA and a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA, wherein each of the first and second micronutrients are added to the composition as a micronutrient supplement.

13. The composition of claim 12, wherein the infant is a preterm infant, preferably a Very Low Birth Weight (VLBW) infant.

14. The composition of claim 12 or 13, wherein the composition further comprises human milk.

15. The composition of claim 14, wherein the human milk is MOM or DHM.

16. The composition of any of claims 12 to 15, wherein the at least two micronutrients are provided as a blend, preferably wherein the blend is a powder, more preferably wherein the powder is provided in a dosage unit selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet.

17. The composition of any of claim 12 to 15, wherein each of the at least two micronutrients is provided separately as a discrete dosage unit wherein: a first dosage unit comprises of a first micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA; and a second dosage unit comprises of a second micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA.

18. The composition of claim 17, wherein the first and second micronutrients are provided as a powder, and preferably the first and second dosage units are selected from the group consisting of a sachet, a stick pack or a dissolvable compressed powder tablet.

19. The composition of any of claims 12 to 18, wherein the at least two micronutrients comprises sodium in an amount of 0.1 to 50 mg, chloride in an amount of 0.1 to 50 mg, zinc in an amount of 0.01 to 5 mg, magnesium in an amount of 0.01 to 5 mg, vitamin A in an amount of 0.01 to 5 mg, vitamin C in an amount of 0.1 to 50 mg, DHA in an amount of 0.1 to 50 mg, and / or inositol in an amount of 0.01 to 50 mg.

20. A human milk fortifier (HMF) micronutrient supplementation kit comprising a single micronutrient, wherein the HMF micronutrient supplementation kit is for use in supplementing the levels of the micronutrient within Mother’s Own Milk (MOM), Donor Human Milk (DHM), a mixture of MOM and HMF (MOM / HMF), and / or within a mixture of DHM and HMF (DHM / HMF), wherein the single micronutrient is selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, inositol and DHA.

21. The HMF micronutrient supplementation kit of claim 20, wherein the single micronutrient is provided as a discrete dosage unit.

22. The HMF micronutrient supplementation kit of claim 21 , wherein the dosage unit comprises a plurality of dosage units wherein each of the dosage units comprises of a micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol.

23. The HMF micronutrient supplementation kit of claim 21 , wherein the dosage unit comprises a first dosage unit and a second dosage unit, wherein the first and second dosage units comprise the same single micronutrient but in different amounts.

24. The HMF micronutrient supplementation kit of claim 23, wherein the first dosage unit and second dosage unit comprise: a plurality of first dosage units, wherein each of the first dosage units comprises of a micronutrient selected from the group consisting of sodium, chloride, magnesium, zinc, vitamin A, vitamin C, DHA and inositol, and a plurality of second dosage units, wherein each of the second dosage units comprises of the same micronutrient as each of the first dosage units.

25. The HMF micronutrient supplementation kit of any of claims 20 to 24, wherein the single micronutrient comprises sodium in an amount of 0.1 to 50 mg, chloride in an amount of 0.1 to 50 mg, zinc in an amount of 0.01 to 5 mg, magnesium in an amount of 0.01 to 5 mg, vitamin A in an amount of 0.01 to 5 mg, vitamin C in an amount of 0.1 to 50 mg, DHA in an amount of 0.1 to 50 mg, or inositol in an amount of 0.01 to 50 mg.

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