PROCESSES FOR THE PRODUCTION OF IRON MICROCAPSULES AND FORTIFIED DAIRY BEVERAGES, PRODUCTS AND USES

Description

1 / 10 “PROCESSES FOR THE PRODUCTION OF IRON MICROCAPSULES AND FORTIFIED DAIRY BEVERAGES, PRODUCTS AND USES”

[01] The technology deals with a process for microencapsulation of ferrous sulfate heptahydrate by the complex coacervation method using whey protein isolate and gum arabic as wall components. It also deals with the manufacturing process of a fermented milk beverage fortified with iron microcapsules, in order to increase the bioavailability of this mineral for the consumer. It also refers to the use of this beverage as food and / or ingredient in the production of other food products for the prevention and treatment of childhood malnutrition.

[02] Recent research indicates that 144 million children under 5 years of age suffered from nutritional atrophy in 2019, equivalent to 21.3% of this segment of the world population (FAO; IFAD; UNICEF; WFP; WHO, 2022. (FAO; IFAD; UNICEF; WFP; WHO. The State of Food Security and Nutrition in the World 2022. [sl])). Similarly, in 2020, there was an increase of 118 million undernourished people, affected by the impact of the pandemic, with the effects extending beyond 2020 and potentially reaching more than 30 million by 2030, in addition to the previous number (GLOBAL NUTRITION REPORT, 2021). Furthermore, in 2021, the number of children suffering from undernourishment increased even further, reaching 149.2 million (FAO; IFAD; UNICEF; WFP; WHO, 2022).

[03] Food fortification with minerals is an effective strategy to combat malnutrition and therefore prevent childhood anemia, providing a more balanced diet for children. The Petition 870240109582, dated 12 / 23 / 2024, p. 10 / 24 2 / 10 Fortified foods should be low-cost, easily accessible to the entire population, and also have high biological value, good stability, and availability for families. Iron is an essential micronutrient for vital metabolic processes. Hence the importance of absorbing it in sufficient quantities so that it performs its function and prevents one of the main diseases caused by mineral deficiency, which is iron-deficiency anemia.

[04] However, fortifying foods with minerals, such as iron, presents a major challenge for the food industry, as the direct addition of iron to food matrices can reduce its stability and bioavailability, as well as cause organoleptic changes in the final product. Iron encapsulation is a relevant alternative to improve the bioavailability of this mineral, in addition to protecting ferrous ions from the gastrointestinal lumen, controlling release kinetics, and masking the unpleasant aroma, color, and taste caused by ferrous ions.

[05] In the state of the art there are several methods for microencapsulation of iron for food fortification. Such as the extrusion method used by Duffy, C et al. (Duffy, C. et al. Preparation of novel chitosan iron microgel beads for fortification applications. Food Hydrocolloids, 84, 608-615, 2018) to fortify infant formula with different iron salts encapsulated using chitosan as a wall material.

[06] Ghorbani, S. et al. (Formulation and assessing characteristics of probiotic ice cream fortified with free and encapsulated iron. Journal of Food Measurement and Characterization, 17, 1, 499-507, 2023) modified the extrusion method to microencapsulate ferrous sulfate in sodium alginate for use of the microcapsules to fortify ice cream. Petition 870240109582, dated 12 / 23 / 2024, page 11 / 24 3 / 10

[07] Sadiq, IH et al. (Sadiq, IH et al. Study of the physicochemical, rheological and sensory properties of yoghurt fortified with microencapsulation iron. Iraqi Journal of Agricultural Sciences, 50, 5, 2019) microencapsulated ferrous sulfate in sodium alginate by the spray dryer method for use of the microcapsules to fortify milk to be used in the production of yogurt.

[08] Gaigher, B. et al. (Gaigher, B. et al. Formulations with microencapsulated Fe-peptides improve in vitro bioaccessibility and bioavailability. Current Research in Food Science, 5, 687-697, 2022.) reveals the use of microencapsulated iron using the spray dryer method with maltodextrin and polydextrin to fortify dry chocolate, strawberry, and tangerine drinks.

[09] Among the different methods of mineral encapsulation, complex coacervation consists of the electrostatic interaction, generally, between a protein and a polysaccharide. Document BR1020220204314, entitled “Process for obtaining zinc microcapsules, process for obtaining fortified dairy beverage, product and uses”, whose priority date is 07 / 13 / 2020, discloses a zinc microencapsulation process by the complex coacervation method using gelatin and gum arabic as wall components. The difference between the two technologies lies in the protein used as wall material, which in the present invention was whey protein isolate. The microencapsulated active core also differs; in this invention, it is iron. Furthermore, the microencapsulation method of the present invention involves the preparation of aqueous and oily phases, in which the proportion of active core is greater than the proportion of wall material, unlike the aforementioned document.

[10] However, no studies using complex coacervation for iron encapsulation were found in the state of the art. Petition 870240109582, dated 12 / 23 / 2024, page 12 / 24 4 / 10 using a protein and a polysaccharide as wall materials. Similarly, no reports were found of the fortification of fermented milk beverages with microencapsulated iron using the complex coacervation method, employing protein and polysaccharide as encapsulating materials. DETAILED DESCRIPTION OF THE TECHNOLOGY

[11] The technology deals with a process for microencapsulation of ferrous sulfate heptahydrate by the complex coacervation method using whey protein isolate and gum arabic as wall components. It also deals with the manufacturing process of a fermented milk beverage fortified with iron microcapsules, in order to increase the bioavailability of this mineral for the consumer. It also refers to the use of this beverage as food and / or ingredient in the production of other food products for the prevention and treatment of childhood malnutrition.

[12] The process of obtaining iron microcapsules comprises the following steps: a. Prepare the external aqueous phase by dissolving ferrous sulfate heptahydrate in water at a concentration of 1030% w / v; b. Prepare the oil phase by mixing an emulsifier selected from the group comprising soy lecithin, sodium alginate, glycerol in oil, the oil being selected from the group comprising soybean oil, corn oil, in an emulsifier:oil ratio of 0.1:99.9-0.15:99.85 w / w; c. Maintain the mixture obtained in step “b” under magnetic stirring at 180-200 rpm for 25-30 minutes at a temperature between 35-40°C; d. Let the mixture obtained in step “c” rest for 25-30 minutes; Petition 870240109582, dated 12 / 23 / 2024, page 13 / 24 5 / 10 e. Prepare a simple emulsion by dispersing, drop by drop, the external aqueous phase obtained in step “a” into the oily phase obtained in step “d”, using a metal disperser coupled to a mechanical propeller stirrer at 5,000 to 10,000 rpm, for 4 to 8 minutes; f. Prepare the internal aqueous phase by dissolving a selected protein from the group comprising albumin, gelatin, whey protein isolate, in water under magnetic stirring, at a temperature of 38-42°C, at a concentration of 1-3% w / v; g. Prepare a double emulsion by dispersing the single emulsion obtained in “e” in the internal aqueous phase obtained in “f” using a metal disperser coupled to a mechanical propeller stirrer at 5,000 to 10,000 rpm for 4 to 8 minutes; h. Prepare a solution of a polysaccharide selected from the group comprising sodium alginate, starch, dextrin, gum arabic, guar gum, xanthan gum, maltodextrin, chitosan, in water at a concentration between 0.5-1.5% w / v; i. Coacervate the polysaccharide solution obtained in “h” by adding it drop by drop to the double emulsion obtained in “g”; j. Adjust the pH obtained in “i” to 3.8-4.2 and adjust the temperature to 38-42°C; k. Cool the mixture obtained in “j” to room temperature; l. Decant the substance obtained in “k”, discard the supernatant and recover the decanted coacervate; m. Freeze the coacervate obtained in “l”; n. Dry the product obtained in “m” by freeze-drying to obtain dry microcapsules. Petition 870240109582, dated 12 / 23 / 2024, page 14 / 24 6 / 10

[13] The process for obtaining iron microcapsules is characterized by the oily phase being prepared in step “b” by mixing 0.1-0.15% (w / w) soy lecithin in 99.85-99.9% (w / w) soybean oil.

[14] The process of obtaining iron microcapsules is characterized by the aqueous phase being whey protein isolate in water 1-3% (m / v) in step “f”.

[15] The process for obtaining iron microcapsules is characterized by the polysaccharide solution in water being a 0.5-1.0% (w / v) solution of gum arabic in step “h”.

[16] The process of obtaining iron microcapsules is characterized by comprising encapsulating materials, a protein and a polysaccharide, and as core material, a form of iron.

[17] The iron microcapsules product is characterized by being produced by the complex coacervation method, using as encapsulating materials, 1.5% w / v whey protein isolate, 1% w / v gum arabic and 30% w / v ferrous sulfate heptahydrate.

[18] The process of obtaining milk beverage fortified with iron microcapsules is characterized by comprising the following steps: a. Mix whole milk (40-50%) and whey (50-60%); b. Add to the mixture obtained in “a” modified starch (0.8-1.0% w / v) and sugar (10-16% w / v); c. Heat the mixture obtained in “b” to 85-90°C for 10-12 minutes; d. Cool the mixture obtained in step “c” to a temperature between 40 and 45°C; e. Add to the mixture obtained in “d” a thermophilic lactic culture (1-1.5% w / v), selected from the group Lactobacillus and Streptococcus; Petition 870240109582, dated 12 / 23 / 2024, p. 15 / 24 7 / 10 f. Incubate the mixture obtained in “e” at 40-44°C for 4-8 hours; g. Cool the mixture obtained in “f” until it reaches a temperature between 5 and 10°C; h. Agitate the clot formed in “g” with a harp-type agitator at 25-30 rpm; I. Add, for each 1L of the mixture produced in “h”, 25% (m / v) of iron microcapsules; j. Bottle the beverage produced in “i” and store under refrigeration at a temperature of 4 to 5°C.

[19] The fortified dairy drink consists of 40-50% whole milk, 50-60% whey, 0.8-1% modified starch, 10-16% sugar; 1-1.5% of selected lactic culture from the Lactobacillus and Streptococcus group, characterized by comprising ferrous sulfate emulsified in soybean oil and soy lecithin and microencapsulated with whey protein isolate and gum arabic, in the proportion of 25% (m / v).

[20] The use of the fortified milk drink product is characterized as being used as food and / or as an ingredient in the preparation of other food products for the prevention and treatment of childhood malnutrition.

[21] The present invention can be better understood through the following examples, which are not limiting. EXAMPLE 1 - PRODUCTION PROCESS OF FERROUS SULFATE MICROCAPSULES

[22] The method used for the production of ferrous sulfate microcapsules employed whey protein isolate and gum arabic as wall materials, through complex coacervation. Encapsulation occurred in a double emulsion process, followed by cooling and freeze-drying. Initially, to prepare the external aqueous phase, ferrous sulfate heptahydrate was solubilized with a Petition 870240109582, dated 12 / 23 / 2024, page 16 / 24 8 / 10 concentration of 30% (w / v) in distilled water. Next, the oil phase was prepared by mixing 0.13% (w / w) soy lecithin in 99.87% (w / w) soybean oil. The mixture was kept under magnetic stirring at 200 rpm for 30 minutes at room temperature. Following the preparation of the phases, the simple emulsion was prepared, which corresponded to the dropwise dispersion of the external aqueous phase in the oil phase. The preparation of the double emulsion corresponded to the dispersion of the simple emulsion in the internal aqueous phase, which was prepared by dissolving whey protein isolate, the encapsulating material, in water at a concentration of 1.5% w / v with a magnetic stirrer at 200 rpm. The dispersions were carried out with a metal disperser coupled to a microgrinder (Dremel 3000) at 10,000 rpm for 4 minutes. Coacervation occurred with the slow addition of gum arabic solution (1% w / v) to the double emulsion. The pH was adjusted to 4 with 1 M HCl at a temperature of 40 °C.The coacervate was slowly cooled to 10 °C in an ice bath and subsequently left to stand for 24 hours at ± 8 °C for decantation. After phase separation, the supernatant was discarded, and the coacervates were frozen and subsequently dried by lyophilization. The microencapsulation technique described in this technology involves the electrostatic interaction between two polymers with opposite charges. In this case, coacervation is only possible if the pH values ​​are below the isoelectric point (pI) of whey protein isolate and above the dissociation constant (pKa) of gum arabic. The pI of whey protein isolate is approximately 5, meaning it has a positive charge at pH 5.<pl. A pKa da goma arábica é próxima de 3,5, ou seja, tem grupos carregados negativamente em pH >pKa. At the pH value used, whey protein isolate becomes positively charged and gum arabic negatively charged, thus forming coacervates.The pH value is therefore a crucial factor and... Petition 870240109582, dated 12 / 23 / 2024, page 17 / 24 9 / 10 limiting factor for microcapsule production using the complex coacervation technique. EXAMPLE 2 - PROCESS FOR OBTAINING A DAIRY BEVERAGE FORTIFIED WITH IRON MICROCAPSULES

[23] For the production of the dairy beverage, a mixture containing 40% v / v whole milk and 60% v / v whey was used. Modified starch (0.8% w / v) and sucrose (10% w / v); thermophilic lactic culture (1% w / v) and microencapsulated ferrous sulfate (25% w / v) were added to this mixture. All ingredients, except iron, were mixed, placed in an industrial yogurt maker and heated to 85 °C for 10 min. The mixture was then cooled to 45 °C, at which point the 1-1.5% w / v lactic culture was added. When the pH reached 4.6, the mixture was cooled and maintained at 5 ± 2 °C throughout the curd breakdown process. According to the Brazilian National Health Surveillance Agency (ANVISA), to be classified as a fortified food, the liquid product must contain 15% of the Recommended Daily Intake (RDI) of iron per 100 mL.Therefore, for every 100 mL of the milk beverage, 2.5% (w / v) of iron microcapsules were added to reach an average value of 1.65 mg of iron / 100 mL of milk beverage, which represents 15% of the highest Recommended Daily Intake (RDA) of iron for children aged 0 to 5 years. This value was estimated based on the RDA of iron (for early childhood) recommended by ANVISA (Brazilian Health Regulatory Agency), the Food and Drug Administration (FDA / USA), and the European Food Safety Authority (EFSA / Europe). The milk beverages were packaged in 200 mL plastic containers and stored under refrigeration at 5 °C ± 1 °C.

[24] The developed technique has good encapsulation efficiency (up to 56%), good yield (up to 61%), and is practical and easy to handle. In addition, the use of microencapsulated iron provides stability during processing and shelf life. Petition 870240109582, dated 12 / 23 / 2024, page 18 / 24 10 / 10 of the final product and during processing, it masks the unpleasant sensory characteristics of iron sulfate. Petition 870240109582, dated 12 / 23 / 2024, page 19 / 24

Claims

1 / 4 CLAIMS 1. PROCESS FOR OBTAINING IRON MICROCAPSULES, characterized by comprising the following steps: a. Preparing the external aqueous phase by dissolving iron sulfate heptahydrate in water at a concentration of 1030% w / v; b. Preparing the oily phase by mixing an emulsifier selected from the group comprising soy lecithin, sodium alginate, glycerol in oil, the oil being selected from the group comprising soybean oil, corn oil, in an emulsifier:oil ratio of 0.1:99.9-0.15:99.85 w / w; c. Maintaining the result obtained in “b” under magnetic stirring at 180-200 rpm for 25-30 minutes at a temperature between 35-40°C; d. Maintaining the result obtained in “c” at rest for 25-30 minutes; e. Prepare a simple emulsion by dispersing, drop by drop, the external aqueous phase obtained in step “a” into the oily phase obtained in step “d”, using a metal disperser coupled to a mechanical propeller stirrer at 5,000 to 10,000 rpm, for 4 to 8 minutes; f.Prepare the internal aqueous phase by solubilizing a selected protein from the group comprising albumin, gelatin, whey protein isolate, in water under magnetic stirring, at a temperature of 38-42°C, at a concentration of 1-3% w / v; g. Prepare a double emulsion by dispersing the single emulsion obtained in “e” in the internal aqueous phase obtained in “f” with a metal disperser coupled to a mechanical propeller stirrer at 5,000 to 10,000 rpm, for 4 to 8 minutes; Petition 870240109582, dated 12 / 23 / 2024, page 20 / 24 2 / 4 h. Prepare a solution of a polysaccharide selected from the group comprising sodium alginate, starch, dextrin, gum arabic, guar gum, xanthan gum, maltodextrin, chitosan, in water at a concentration between 0.5-1.5% w / v; i. Coacervate the polysaccharide solution obtained in “h” by adding it dropwise to the double emulsion obtained in “g”; j. Adjust the pH of the solution obtained in “i” to 3.8-4.2 and adjust the temperature to 38-42°C; k. Cool the solution obtained in “j” to room temperature; l.Decant the material obtained in step “k”, discard the supernatant and recover the decanted coacervate; m. Freeze the coacervate obtained in step “l”; n. Dry the material obtained in step “m” by lyophilization to obtain the dry microcapsules.

2. PROCESS, according to claim 1, characterized in that, in step “b”, the oil phase is prepared by mixing 0.1-0.15% (w / w) of soy lecithin in 99.85-99.9% (w / w) of soybean oil.

3. PROCESS, according to claim 1, characterized in that, in step “f”, the aqueous phase is whey protein isolate in water 1-3% (w / v).

4. PROCESS, according to claim 1, characterized in that, in step “h”, the polysaccharide solution in water is a 0.5-1% (w / v) gum arabic solution.

5. IRON MICROCAPSULES PRODUCT obtained as defined in any one of claims 1 to 5, characterized by comprising encapsulating materials, a protein from the group comprising albumin, gelatin, whey protein isolate, in water, at a concentration of 1.5% (w / v); a polysaccharide from the group Petition 870240109582, dated 12 / 23 / 2024, page 21 / 24 3 / 4 comprising sodium alginate, starch, gum arabic, guar gum, xanthan gum, maltodextrin, chitosan, at a concentration of 1% (w / v); and as core material, ferrous sulfate hephihydrate at a concentration of 30% (w / v).

6. PROCESS FOR OBTAINING A FORTIFIED DAIRY BEVERAGE, using the iron microcapsules defined in any of claims 6 and 7, characterized by comprising the following steps: a. Mixing whole milk (40-50%) and whey (50-60%); b. Adding modified starch (0.8-1.0% w / v) and sugar (10-16% w / v) to the mixture obtained in “a”; c. Heating the mixture obtained in “b” to 85-90°C for 10-12 minutes; d. Cooling the mixture obtained in “c” to a temperature between 40 and 45°C; e. Adding a thermophilic lactic culture (1-1.5% w / v), selected from the Lactobacillus and Streptococcus groups, to the mixture obtained in “d”; f. Incubating the mixture obtained in “e” at 40-44°C for 4-8 hours; g. Cool the mixture obtained in “f” until it reaches a temperature between 5 and 10°C; h. Stir the curd formed in “g” with a harp-type stirrer at 25-30 rpm; i. Add, for each 1L of the mixture produced in “h”, 25% (m / v) of iron microcapsules; j.Bottle the beverage produced in container “i” and store under refrigeration at a temperature of 4 to 5°C.

7. FORTIFIED DAIRY BEVERAGE PRODUCT, obtained by the process defined in claim 8, consisting of 40-50% whole milk, 50-60% whey, 0.8-1% modified starch, 10-16% sugar; 1-1.5% of selected lactic culture from the Lactobacillus and Streptococcus group, characterized by comprising ferrous sulfate emulsified in soybean oil and soy lecithin and microencapsulated with whey protein isolate and gum arabic, in a proportion of 25% (m / v).

8. USE OF THE PRODUCTS defined in any of claims 6, 7 and 9, characterized by being in the preparation of food products for the prevention and treatment of childhood malnutrition. Petition 870240109582, dated 12 / 23 / 2024, pp. 23 / 24