Polysaccharide-protein composite stable high-bioavailability ferrous microemulsion as well as preparation method and application thereof

Abstract

The invention relates to the field of preparation of micro-emulsions, in particular to a polysaccharide-protein composite stable high-bioavailability ferrous micro-emulsion as well as a preparation method and application thereof. The preparation method of the polysaccharide-protein composite stable high-bioavailability ferrous microemulsion comprises the following steps: adding mineral substances into water, mixing, and carrying out first stirring treatment to obtain a core material solution; adding starch sodium octenylsuccinate, whey protein isolate and polysaccharides into water, mixing, and sequentially carrying out heating treatment and second stirring treatment to obtain a wall material solution; mixing the core material solution, the wall material solution and an emulsifier, and sequentially carrying out third stirring treatment and homogeneous emulsification treatment to obtain an emulsion; and carrying out spray drying treatment on the emulsion to obtain the product. The polysaccharide-protein compounded stable high-bioavailability ferrous microemulsion has excellent embedding rate and storage stability, can effectively protect ferrous ions and reduce the generation of ferric ions, and is beneficial to improving the bioavailability of ferrous iron.

Description

Technical Field

[0001] The present invention relates to the field of preparation of microemulsions, and particularly to a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, a preparation method thereof, and an application thereof. Background Art

[0002] Iron is one of the essential trace elements in the human body. In the human body, it mainly participates in the synthesis of hemoglobin in the form of ferrous (Fe2+), promotes the transportation of oxygen, and the energy metabolism of cells, etc. Iron deficiency may lead to health problems such as anemia and decreased immunity, especially more common in pregnant women, infants, and some special populations.

[0003] At present, the prior art usually uses ferrous sulfate, ferrous gluconate, ferrous fumarate, etc. as raw materials to prepare iron supplements. However, during the storage process of the iron supplements prepared by traditional methods, the ferrous (Fe 2+ ) is extremely easy to be oxidized to ferric iron (Fe3+), resulting in a decrease in the bioavailability of ferrous. Summary of the Invention

[0004] The present invention provides a preparation method of a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion. Through this method, a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion with excellent encapsulation efficiency and storage stability can be prepared. The polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion can effectively protect ferrous ions, reduce the generation of ferric ions, and improve the bioavailability of ferrous.

[0005] The present invention also provides a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, which is prepared by the above preparation method. Therefore, this microemulsion has excellent encapsulation efficiency and storage stability, can effectively protect ferrous ions, reduce the generation of ferric ions, and improve the bioavailability of ferrous.

[0006] The present invention also provides an application of the above polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion in the preparation of iron supplements. Through research by the inventors of the present invention, it is shown that the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion has excellent encapsulation efficiency and storage stability, can effectively protect ferrous ions, reduce the generation of ferric ions, and improve the bioavailability of ferrous; and through animal experiments, it is shown that using this polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion can significantly increase the titanium content in the serum of mice, indicating that using this polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion can greatly improve the absorption and utilization of ferrous and effectively improve the bioavailability of ferrous; therefore, this polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion can be used to prepare iron supplements.

[0007] The first aspect of the present invention provides a method for preparing a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, comprising the following steps:

[0008] Mix minerals in water and obtain a core material solution through a first stirring treatment;

[0009] Mix sodium octenyl succinate starch, whey protein isolate and polysaccharides in water, and obtain a wall material solution through a heating treatment and a second stirring treatment in sequence;

[0010] Mix the core material solution, the wall material solution and an emulsifier, and obtain an emulsion through a third stirring treatment and a homogenization emulsification treatment in sequence;

[0011] Perform spray drying treatment on the emulsion to obtain the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion;

[0012] The minerals are at least one of ferrous sulfate heptahydrate and ferrous gluconate.

[0013] In the method for preparing the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion as described above, in the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, the mass ratio of minerals, sodium octenyl succinate starch, whey protein isolate, polysaccharides and emulsifier is (1-20):(3-20):(1-15):(0.5-5):(1-2).

[0014] In the method for preparing the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion as described above, the polysaccharides are at least one of chitosan and sucrose;

[0015] And / or, the emulsifier is at least one of mono- and diglycerol fatty acid esters and Tween-80.

[0016] In the method for preparing the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion as described above, the pressure of the homogenization emulsification treatment is 200 bar to 700 bar, the temperature is 40 °C to 45 °C, and the number of times of the homogenization emulsification treatment is 2 to 6 times.

[0017] In the method for preparing the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion as described above, the rotation speed of the first stirring treatment is 200 rpm to 500 rpm, and the time is 50 min to 100 min.

[0018] In the method for preparing the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion as described above, the rotation speed of the second stirring treatment is 200 rpm to 300 rpm, and the time is 20 min to 50 min.

[0019] The preparation method of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion as described above, wherein the temperature of the heat treatment is 45°C to 55°C.

[0020] The preparation method of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion as described above, wherein the emulsion is subjected to spray drying treatment, including: using a spray dryer to perform spray drying treatment on the emulsion, setting the inlet air temperature of the spray dryer to 175°C to 185°C, the outlet air temperature to 80°C to 90°C, the centrifugal rotation speed to 250 rpm to 300 rpm, and the feeding rotation speed to 15 rpm to 20 rpm.

[0021] The second aspect of the present invention provides a polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion, which is prepared by the preparation method of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion as described above.

[0022] The third aspect of the present invention provides an application of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion as described above in the preparation of iron supplements.

[0023] Compared with the prior art, the solution of the present invention has at least the following effects:

[0024] The preparation method of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion provided by the present invention uses minerals (the minerals are at least one of ferrous sulfate heptahydrate and ferrous gluconate), sodium octenyl succinate starch, whey protein isolate, polysaccharides, and emulsifiers as raw materials. Through this method, a polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion with excellent encapsulation efficiency and storage stability can be prepared. This polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion can effectively protect ferrous ions, reduce the generation of ferric ions, and is beneficial to improving the bioavailability of ferrous. Detailed Embodiments

[0025] To make the objectives, technical solutions, and advantages of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0026] In the embodiments of the present invention, where specific technologies or conditions are not indicated, they shall be carried out according to the technologies or conditions described in the literature in the field or according to the product specifications. Reagents or instruments not indicated by the manufacturer can all be conventional products obtained through commercial purchase.

[0027] It should be noted that the descriptions involving "first", "second", etc. in the present invention are used to distinguish similar objects, rather than to describe a specific order or sequence, and thus should not be construed as a limitation to the present invention.

[0028] The first aspect of the present invention provides a method for preparing a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, comprising the following steps:

[0029] Mix the mineral in water and obtain the core material solution through the first stirring treatment;

[0030] Mix sodium starch octenyl succinate, whey protein isolate and polysaccharides in water, and obtain the wall material solution through heating treatment and the second stirring treatment in sequence;

[0031] Mix the core material solution, the wall material solution and the emulsifier, and obtain the emulsion through the third stirring treatment and the homogenization emulsification treatment in sequence;

[0032] Perform spray drying treatment on the emulsion to obtain the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion;

[0033] The mineral is at least one of ferrous sulfate heptahydrate and ferrous gluconate.

[0034] The present invention does not make special limitations on the specific sources of the above raw materials, and they can be obtained through commercial channels.

[0035] The present invention does not make special limitations on the rotation speed and time of the above third stirring treatment, and can be selected according to actual needs.

[0036] The object of the present invention for preparation is a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion. Specifically, minerals are added to water and mixed to obtain a first mixture, and the first mixture is subjected to a first stirring treatment to obtain a core material solution. The first stirring treatment is to make the minerals and water fully and evenly mixed; sodium octenyl succinate starch, whey protein isolate, and polysaccharides are added to water and mixed to obtain a second mixture, and the second mixture is successively subjected to a heating treatment and a second stirring treatment to obtain a wall material solution. The heating treatment is to make sodium octenyl succinate starch, whey protein isolate, and polysaccharides fully dissolve in water without damaging each component, and the second stirring treatment is to make sodium octenyl succinate starch, whey protein isolate, and polysaccharides fully and evenly mixed with water; the core material solution, the wall material solution, and an emulsifier are mixed to obtain a third mixture, and the third mixture is successively subjected to a third stirring treatment and a homogenization emulsification treatment to obtain an emulsion. The third stirring treatment is to make the core material solution, the wall material solution, and the emulsifier fully and evenly mixed, and the homogenization emulsification treatment is to improve the dispersibility of ferrous ions in the emulsion, which is beneficial to the subsequent preparation of a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion with excellent storage stability; the emulsion is subjected to spray drying treatment to quickly evaporate the water on the surface of the emulsion through heat exchange, so that ferrous ions are effectively encapsulated and the oxidation of ferrous ions is reduced, thereby obtaining a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion.

[0037] Through the above method, the present invention can prepare a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion that can effectively protect ferrous ions and has excellent encapsulation rate and storage stability.

[0038] In a specific embodiment, in the above polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, the mass ratio of minerals, sodium octenyl succinate starch, whey protein isolate, polysaccharides, and emulsifier is (1-20):(3-20):(1-15):(0.5-5):(1-2). For example, the mass ratio of minerals, sodium octenyl succinate starch, whey protein isolate, polysaccharides, and emulsifier is 1:3:1:0.5:1, 20:20:15:5:2, 9:5:2:1:1.5, 9:3:1:0.5:1, etc.

[0039] When the parameters of the mass ratio of minerals, sodium octenyl succinate starch, whey protein isolate, polysaccharides, and emulsifier in the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion are within the above range, each substance can be fully matched, and a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion with excellent encapsulation rate and storage stability can be prepared. The polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion can effectively protect ferrous ions and reduce the generation of ferric ions.

[0040] In a specific embodiment, the above polysaccharides are at least one of chitosan and sucrose.

[0041] When the polysaccharides are the above substances, the polysaccharides can crosslink or electrostatically adsorb with sodium octenyl succinate starch and whey protein isolate, thereby increasing the thickness of the wall material, more effectively protecting ferrous ions, and preventing the oxidation of ferrous ions.

[0042] In a specific embodiment, the above emulsifier is at least one of mono- and diglycerol fatty acid esters and Tween-80.

[0043] When the emulsifier is the above substance, it can maintain the stability of the emulsion, so that after the emulsion is spray-dried, a high-bioavailability ferrous microemulsion with polysaccharide-protein composite stability can be obtained, which can effectively protect ferrous ions and has excellent entrapment efficiency and storage stability.

[0044] In a specific embodiment, the pressure of the above homogenization and emulsification treatment is 200 bar to 700 bar, the temperature is 40 °C to 45 °C, and the number of times of homogenization and emulsification treatment is 2 to 6 times.

[0045] When the parameters of the pressure, temperature, and number of times of homogenization and emulsification treatment are each within the above ranges, ferrous ions can be evenly dispersed in the emulsion, and the entrapment efficiency of ferrous ions can be improved.

[0046] Exemplarily, the pressure of the above homogenization and emulsification treatment can be any one of 200 bar, 300 bar, 400 bar, 500 bar, 600 bar, 700 bar or a range composed of any two of them;

[0047] The temperature can be any one of 40 °C, 41 °C, 42 °C, 43 °C, 44 °C, 45 °C or a range composed of any two of them;

[0048] The number of times of homogenization and emulsification treatment can be any one of 2 times, 3 times, 4 times, 5 times, 6 times or a range composed of any two of them.

[0049] In a specific embodiment, the rotation speed of the above first stirring treatment is 200 rpm to 500 rpm, and the time is 50 min to 100 min.

[0050] When the parameters of the rotation speed and time of the first stirring treatment are each within the above ranges, the minerals and water are more fully mixed and homogenized.

[0051] Exemplarily, the rotation speed of the above first stirring treatment can be any one of 200 rpm, 250 rpm, 300 rpm, 350 rpm, 400 rpm, 450 rpm, 500 rpm or a range composed of any two of them;

[0052] The time can be any one of 50 min, 60 min, 70 min, 80 min, 90 min, 100 min or a range composed of any two of them.

[0053] In a specific embodiment, the rotation speed of the above-mentioned second stirring treatment is 200 rpm to 300 rpm, and the time is 20 min to 50 min.

[0054] When the parameters of the rotation speed and time of the second stirring treatment are respectively within the above ranges, sodium octenyl succinate starch, whey protein isolate and polysaccharides are fully mixed and homogenized with water.

[0055] Exemplarily, the rotation speed of the above-mentioned second stirring treatment can be any one of 200 rpm, 250 rpm, 300 rpm or a range composed of any two of them;

[0056] The time can be any one of 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min or a range composed of any two of them.

[0057] In a specific embodiment, the temperature of the above-mentioned heating treatment is 45 °C to 55 °C.

[0058] The present invention does not particularly limit the time of the heating treatment, as long as sodium octenyl succinate starch, whey protein isolate and polysaccharides can be fully dissolved in water.

[0059] When the parameter of the temperature of the heating treatment is within the above range, sodium octenyl succinate starch, whey protein isolate and polysaccharides are more fully dissolved in water without damaging each component.

[0060] Exemplarily, the temperature of the above-mentioned heating treatment can be any one of 45 °C, 46 °C, 47 °C, 48 °C, 49 °C, 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C or a range composed of any two of them.

[0061] In a specific embodiment, the above-mentioned spray drying treatment of the emulsion includes: using a spray dryer to perform spray drying treatment on the emulsion, setting the inlet air temperature of the spray dryer to 175 °C to 185 °C, the outlet air temperature to 80 °C to 90 °C, the centrifugal rotation speed to 250 rpm to 300 rpm, and the feeding rotation speed to 15 rpm to 20 rpm.

[0062] The present invention performs spray drying treatment on the emulsion through the above process. When the parameters of the inlet air temperature, outlet air temperature, centrifugal rotation speed, and feed rotation speed of the spray dryer are within the above ranges respectively, the water on the surface of the emulsion is quickly evaporated through heat exchange, the ferrous ions are effectively embedded, the oxidation of ferrous ions is reduced, which is beneficial to the preparation of a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion that can effectively protect ferrous ions and has excellent embedding rate and storage stability.

[0063] Exemplarily, the above inlet air temperature can be any one of 175°C, 176°C, 177°C, 178°C, 179°C, 180°C, 181°C, 182°C, 183°C, 184°C, 185°C or the range composed of any two of them;

[0064] The outlet air temperature can be any one of 80°C, 81°C, 82°C, 83°C, 84°C, 85°C, 86°C, 87°C, 88°C, 89°C, 90°C or the range composed of any two of them;

[0065] The centrifugal rotation speed can be any one of 250 rpm, 260 rpm, 270 rpm, 280 rpm, 290 rpm, 300 rpm or the range composed of any two of them;

[0066] The feed rotation speed can be any one of 15 rpm, 16 rpm, 17 rpm, 18 rpm, 19 rpm, 20 rpm or the range composed of any two of them.

[0067] The second aspect of the present invention provides a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion, which is prepared by the preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion described above. Therefore, this microemulsion has excellent embedding rate and storage stability, can effectively protect ferrous ions, reduce the generation of ferric ions, and improve the bioavailability of ferrous.

[0068] The third aspect of the present invention provides an application of the above polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion in the preparation of iron supplements.

[0069] The present invention will be described in detail for its embodiments through the following examples and comparative examples.

[0070] Example 1 (using ferrous sulfate heptahydrate and ferrous gluconate as raw materials)

[0071] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example includes the following steps:

[0072] (1) By mass parts, 5 parts of ferrous sulfate heptahydrate and 4 parts of ferrous gluconate are added to 50 parts of water and mixed to obtain a first mixture. Then, the first mixture is stirred at 300 rpm for 60 min to obtain a core material solution.

[0073] (2) By mass parts, 5 parts of sodium octenyl succinate starch, 2 parts of whey protein isolate, and 1 part of chitosan are added to 50 parts of water and mixed to obtain a second mixture. Then, the second mixture is heated in a water bath at 50 °C to obtain the second mixture after heat treatment. The second mixture after heat treatment is stirred at 200 rpm for 30 min to obtain a wall material solution.

[0074] (3) By mass parts, the core material solution in (1), the wall material solution in (2), 1 part of mono- and diglycerol fatty acid esters, and 0.5 part of Tween-80 are mixed to obtain a third mixture. After the third mixture is stirred evenly, the stirred third mixture is subjected to a homogenization and emulsification treatment under a high-pressure homogenizer. The pressure of the homogenization and emulsification treatment is 550 bar, the temperature is 45 °C, and the number of times of the homogenization and emulsification treatment is 3 times to obtain an emulsion.

[0075] (4) The emulsion is subjected to a spray drying treatment using a spray dryer. The inlet air temperature of the spray dryer is set at 180 °C, the outlet air temperature is 85 °C, the centrifugal rotation speed is 280 pm, and the feeding rotation speed is 18 rpm, thus obtaining a polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion.

[0076] Example 2 (using ferrous sulfate heptahydrate as the raw material)

[0077] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0078] (1) By mass parts, 9 parts of ferrous sulfate heptahydrate are added to 50 parts of water and mixed to obtain a first mixture. Then, the first mixture is stirred at 300 rpm for 60 min to obtain a core material solution.

[0079] Example 3 (using ferrous gluconate as the raw material)

[0080] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0081] (1) By mass parts, 9 parts of ferrous gluconate are added to 50 parts of water and mixed to obtain a first mixture. Then, the first mixture is stirred at 300 rpm for 60 min to obtain a core material solution.

[0082] Example 4

[0083] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0084] (3) The pressure for the homogenization and emulsification treatment is 200 bar.

[0085] Example 5

[0086] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0087] (3) The pressure for the homogenization and emulsification treatment is 700 bar.

[0088] Example 6 (without adding Tween-80)

[0089] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0090] (3) By mass, mix the core material solution in (1), the wall material solution in (2) and 1 part of mono- and diglycerol fatty acid esters to obtain a third mixture. After uniformly stirring the third mixture, obtain the stirred third mixture. Place the stirred third mixture under a high-pressure homogenizer for homogenization and emulsification treatment. The pressure for the homogenization and emulsification treatment is 550 bar, the temperature is 45 °C, and the number of times of the homogenization and emulsification treatment is 3 times to obtain an emulsion.

[0091] Example 7 (without adding mono- and diglycerol fatty acid esters)

[0092] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this example is basically the same as that in Example 1, except that:

[0093] (3) By mass, mix the core material solution in (1), the wall material solution in (2) and 0.5 part of Tween-80 to obtain a third mixture. After uniformly stirring the third mixture, obtain the stirred third mixture. Place the stirred third mixture under a high-pressure homogenizer for homogenization and emulsification treatment. The pressure for the homogenization and emulsification treatment is 550 bar, the temperature is 45 °C, and the number of times of the homogenization and emulsification treatment is 3 times to obtain an emulsion.

[0094] Comparative Example 1

[0095] The preparation method of the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsion provided in this comparative example is basically the same as that in Example 1, except that:

[0096] (2) Replace 5 parts of sodium starch octenyl succinate with 5 parts of gum arabic.

[0097] Comparative Example 2

[0098] The preparation method of the polysaccharide-protein composite-stabilized highly bioavailable ferrous microemulsion provided in this comparative example is basically the same as that of Example 1, except that:

[0099] (2) Replace 5 parts of sodium starch octenyl succinate with 5 parts of gelatin.

[0100] Comparative Example 3 (without adding whey protein isolate)

[0101] The preparation method of the polysaccharide-protein composite-stabilized highly bioavailable ferrous microemulsion provided in this comparative example is basically the same as that of Example 1, except that:

[0102] (2) By mass, add 5 parts of sodium starch octenyl succinate and 1 part of chitosan to 50 parts of water and mix to obtain a second mixture. Then heat the second mixture in a water bath at 50 °C to obtain the heat-treated second mixture. Stir the heat-treated second mixture at 200 rpm for 30 min to obtain the wall material solution.

[0103] Comparative Example 4 (without adding chitosan)

[0104] The preparation method of the polysaccharide-protein composite-stabilized highly bioavailable ferrous microemulsion provided in this comparative example is basically the same as that of Example 1, except that:

[0105] (2) By mass, add 5 parts of sodium starch octenyl succinate and 2 parts of whey protein isolate to 50 parts of water and mix to obtain a second mixture. Then heat the second mixture in a water bath at 50 °C to obtain the heat-treated second mixture. Stir the heat-treated second mixture at 200 rpm for 30 min to obtain the wall material solution.

[0106] Performance test

[0107] (1) Storage stability test experiment

[0108] Place the sample in a sealed container and store it at -18 °C, 4 °C, room temperature, and 45 °C for 180 days. Measure the particle size of the sample at room temperature at the initial state and after 180 days, respectively.

[0109] Take the polysaccharide-protein composite-stabilized highly bioavailable ferrous microemulsions in Examples 1-7 and Comparative Examples 1-4 as samples, and detect the particle size of each sample. The results are shown in Table 1.

[0110] Table 1 Results of particle size stability test

[0111]

[0112] As can be seen from Table 1, the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion in Example 1 of the present invention exhibits good particle size stability. The particle size change ranges of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsions in Example 1, Example 2, Example 3, Example 5, Example 6, Example 7, Example 4, Comparative Example 4, Comparative Example 3, Comparative Example 2, and Comparative Example 1 increase in sequence, and the particle size stabilities decrease in sequence. Among them, the particle size and the particle size change range of the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion in Example 1 are the smallest, indicating that the polysaccharide-protein complex-stabilized high-bioavailability ferrous microemulsion in Example 1 of the present invention has excellent storage stability.

[0113] (2) Encapsulation efficiency test

[0114] Encapsulation efficiency test method:

[0115] Determination of total iron content: Take 50 mg of the sample and place it in a 10 mL centrifuge tube. Add 0.02 g of papain, 0.02 g of amylase, and 1 mL of water. After mixing, heat it in a water bath at 50 °C for 30 min. After taking it out and cooling, adjust the pH = 3 with hydrochloric acid, heat it to boiling for 10 minutes, cool it, add 0.2 mL of a hydrochloric acid hydroxylamine solution with a mass fraction of 10%, shake well, add 0.2 mL of an o-phenanthroline solution with a mass fraction of 0.15% and 0.2 mL of a sodium acetate buffer solution with a concentration of 2 mol / L (adjust the pH to 5.0 with the sodium acetate buffer solution) after 1 minute, and finally make up the volume to 10 mL (constant volume) with water. After 10 minutes, detect the iron ion concentration in the sample at 510 nm with a spectrophotometer, and calculate the total iron content according to the following formula:

[0116] Total iron content = (iron ion concentration × constant volume) / sample mass;

[0117] Determination of the encapsulated iron content: Mix 50 mg of the sample with 10 mL of an ethanol aqueous solution with a mass fraction of 50%, shake well and let stand for 30 minutes, then filter to obtain the solid. After drying the solid, transfer it to a 10 mL centrifuge tube, add 0.02 g of papain, 0.02 g of amylase and 1 mL of water, mix well and heat in a water bath at 50 °C for 30 min. After taking it out and cooling, adjust the pH to 3 with hydrochloric acid, heat and boil for 10 minutes. After cooling, add 0.2 mL of a hydrochloric acid hydroxylamine solution with a mass fraction of 10%, shake well, and then add 0.2 mL of an o-phenanthroline solution with a mass fraction of 0.15% and 0.2 mL of a sodium acetate buffer solution with a concentration of 2 mol / L (adjust the pH to 5.0 with the sodium acetate buffer solution) after 1 minute. Finally, make up the volume to 10 mL with water (the final volume). After 10 minutes, detect the iron ion concentration in the sample by spectrophotometer at 510 nm, and calculate the encapsulated iron content according to the following formula:

[0118] Encapsulated iron content = (iron ion concentration × final volume) / mass of the sample;

[0119] The encapsulation efficiency is obtained by the following formula:

[0120] Encapsulation efficiency = (encapsulated iron content / total iron content) × 100%.

[0121] Take the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsions in Examples 1-7 and Comparative Examples 1-4 of the present invention as samples respectively, and conduct encapsulation efficiency tests on the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsions in Examples 1-7 and Comparative Examples 1-4 of the present invention respectively according to the above encapsulation efficiency test method. The test results are shown in Table 2.

[0122] Table 2 Encapsulation efficiency test results

[0123]

[0124]

[0125] As can be seen from Table 2, compared with the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsions in Comparative Examples 1-4, the polysaccharide-protein composite-stabilized high-bioavailability ferrous microemulsions in Examples 1-7 of the present invention have excellent encapsulation efficiency, with the highest reaching 99.2%.

[0126] (3) Stability test of ferrous ions

[0127] Place 50 g of the sample in a beaker and store it at room temperature for 10 days. At 0 day, 5 days and 10 days of storage respectively, detect the content of ferrous ions and ferric ions in the sample, and then calculate the proportion of the content of ferrous ions and the proportion of the content of ferric ions in the sample respectively through the following formula;

[0128] Content of ferrous ions = Content of ferrous ions / (Content of ferrous ions + Content of ferric ions) × 100%;

[0129] Content of ferric ions = Content of ferric ions / (Content of ferrous ions + Content of ferric ions) × 100%;

[0130] Taking the polysaccharide - protein complex - stabilized highly bio - available ferrous microemulsions in Examples 1 - 7 and Comparative Examples 1 - 4 of the present invention as samples respectively, ferrous ion stability tests were carried out, and the results are shown in Table 3.

[0131] Table 3 Results of the proportion of ferrous ion content and the proportion of ferric ion content

[0132]

[0133]

[0134] As can be seen from Table 3, after storing at room temperature for 10 days, the ferrous ion stability of the polysaccharide - protein complex - stabilized highly bio - available ferrous microemulsions in Examples 1 - 7 of the present invention is better than that of the polysaccharide - protein complex - stabilized highly bio - available ferrous microemulsions in Comparative Examples 1 - 4. Comparing Example 1 with Comparative Examples 1 - 2, it is found that when 5 parts of sodium octenyl succinate starch are replaced by 5 parts of arabic gum or gelatin, after storing at room temperature for 10 days, the content difference of ferrous ions (Fe 2+ ) is large, indicating that obvious oxidation reactions occur when using arabic gum or gelatin, causing more ferrous ions (Fe 2+ ) to be oxidized to ferric ions (Fe 3+ ). Comparing Example 1 of the comparative example with Comparative Examples 3 - 4, it is found that when whey protein isolate or chitosan is not added, after storing at room temperature for 10 days, the content difference of ferrous ions (Fe 2+ ) is relatively large, indicating that the absence of whey protein isolate or chitosan will lead to an exacerbation of the oxidation reaction and an increase in the content of generated ferric ions (Fe 3+ ). The above results show that the polysaccharide - protein complex - stabilized highly bio - available ferrous microemulsion provided in the examples of the present invention has excellent stability, can effectively protect ferrous ions, reduce the generation of ferric ions, and improve the bio - availability of ferrous.

[0135] (4) Animal experiments

[0136] Thirty-six 8-week-old male C57BL / 6 mice were randomly divided into 12 groups (3 mice in each group). The polysaccharide-protein complex-stabilized highly bioavailable ferrous microemulsions in Examples 1-7 and Comparative Examples 1-4 were diluted to a final concentration of 4 mg / L as the experimental groups, and the blank control group was normal saline. They were gavaged once a day for 7 consecutive days, and the gavage volume was calculated at 10 μL / g body weight. Before gavage and on the 7th day, 100 μL of blood was collected from the tail vein of the mice. After heparin anticoagulation, the serum was separated, and the iron content in the mouse serum before gavage and the iron content in the mouse serum on the 7th day were detected by the Ferene method respectively. The detection results are shown in Table 4.

[0137] The increase ratio of iron content = (iron content on the 7th day - iron content on the 0th day) / iron content on the 0th day × 100%, where the iron content on the 0th day refers to the iron content in the mouse serum before gavage, and the iron content on the 7th day refers to the iron content in the mouse serum on the 7th day.

[0138] Table 4 Detection results

[0139]

[0140]

[0141] As can be seen from Table 4, after 7 days, the iron content in the mouse serum of the blank control group (normal saline) only increased by 0.2%, while the use of the polysaccharide-protein complex-stabilized highly bioavailable ferrous microemulsions in Examples 1-7 of the present invention significantly increased the iron content in the mouse serum, with an increase of 34.28% - 68.68%. Among them, the microemulsion in Example 1 had the most significant improvement effect, reaching 68.68%, and its improvement effect was significantly better than that of Comparative Examples 1-4. The above results show that the polysaccharide-protein complex-stabilized highly bioavailable ferrous microemulsion in Example 1 has the best effect on increasing the iron content in the mouse serum, indicating that the use of the polysaccharide-protein complex-stabilized highly bioavailable ferrous microemulsion provided in the examples of the present invention can greatly improve the absorption and utilization of ferrous iron and effectively improve the bioavailability of ferrous iron.

[0142] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions recorded in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability, characterized in that: The following steps are involved: Adding minerals into water and mixing, and obtaining a core material solution through a first stirring process; Sodium starch octenylsuccinate, whey protein isolate and polysaccharides are added into water and mixed, and then subjected to heating treatment and a second stirring treatment in sequence to obtain a wall material solution; The core material solution, the wall material solution and the emulsifier are mixed, and subjected to a third stirring process and a homogenizing emulsification process in sequence to obtain an emulsion; The emulsion is subjected to spray drying treatment to obtain the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability; The mineral is at least one of ferrous sulfate heptahydrate and ferrous gluconate.

2. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: In the polysaccharide-protein composite stabilized high-bioavailability ferrous microemulsion, the mass ratio of minerals, sodium starch octenylsuccinate, whey protein isolate, polysaccharides and emulsifier is (1-20): (3-20): (1-15): (0.5-5): (0.5-2).

3. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The polysaccharide is at least one of chitosan and sucrose; And / or, the emulsifier is at least one of mono- and di-glycerol fatty acid esters and Tween-80.

4. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The pressure of the homogenization and emulsification treatment is 200 bar to 700 bar, the temperature is 40° C. to 45° C., and the number of times of the homogenization and emulsification treatment is 2 to 6 times.

5. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The first stirring treatment is performed at a rotation speed of 200 rpm to 500 rpm and for a time of 50 min to 100 min.

6. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The second stirring treatment has a rotation speed of 200 rpm to 300 rpm and a time of 20 min to 50 min.

7. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The temperature of the heating treatment is 45°C to 55°C.

8. The method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability according to claim 1, characterized in that: The emulsion is spray-dried, comprising: using a spray dryer to spray-dry the emulsion, setting the inlet air temperature of the spray dryer to 175°C-185°C, the outlet air temperature to 80°C-90°C, the centrifugal speed to 250rpm-300rpm, and the feed speed to 15rpm-20rpm.

9. A polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability, characterized in that: The invention is prepared by the method for preparing the polysaccharide-protein composite stabilized ferrous microemulsion with high bioavailability as claimed in any one of claims 1 to 8.

10. Use of the polysaccharide-protein complex-stabilized ferrous microemulsion with high bioavailability according to claim 9 in the preparation of an iron supplement.

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