Strong-absorption water-soluble lutein compound preparation and application thereof in eyesight protection functional food

Abstract

The invention discloses a strong-absorption water-soluble lutein compound preparation and application thereof in food with an eyesight protection function. The xanthophyll-dipotassium glycyrrhizinate water-soluble granules are water-soluble granules formed by molecular self-assembly of xanthophyll, dipotassium glycyrrhizinate, triterpenic acid, protein and dietary polysaccharide. Compared with untreated xanthophyll, the xanthophyll compound preparation with strong absorption has the advantages that the xanthophyll has good water solubility and stability, the intestinal absorption efficiency is improved by 20-35 times, and eyesight protection can be effectively realized. Meanwhile, the embedding rate is high, the safety is good, and the production cost is low.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, and in particular to a highly absorbable water-soluble lutein composite preparation and application thereof in vision-protecting functional foods. Background Art

[0002] Lutein not only plays an important role in promoting the development of infant optic nerves, but also has a variety of functional characteristics such as preventing myopia, preventing and treating age-related macular degeneration, and anti-aging, and has broad application prospects in the food industry. However, as a fat-soluble nutritional active substance, lutein has the disadvantages of poor water solubility, instability under light / heat and acid-base conditions, and bioavailability. Therefore, how to simultaneously improve the solubility, processing stability and intestinal absorption efficiency of lutein has become a key technical problem that needs to be solved in the development of food nutrition preparations and functional foods using lutein. The currently used macromolecule (such as protein, polysaccharide and its complex) embedding technology can solve the processing stability and digestion stability of lutein, and has made a positive contribution to promoting the industrial application of lutein. However, the existing technology involves the use of a large number of chemical reagents, such as the use of chemical emulsifiers such as Tween to dissolve fat-soluble components, and the use of acid-base regulation to promote the interaction between fat-soluble substances and proteins / polysaccharides, which poses a food safety hazard. Therefore, green, safe and efficient processing methods have become key technologies urgently needed for industrialization. In addition, the reason for the low intestinal absorption efficiency of lutein is not only due to its poor stability during digestion, but more importantly, lutein is strictly regulated by lipid transport proteins during trans-intestinal epithelial transport. Therefore, how to effectively regulate intestinal epithelial transport proteins on the basis of encapsulation processing to promote the efficient absorption of lutein has become another technical bottleneck problem that the industry needs to solve urgently. Summary of the invention

[0003] Aiming at the industrial application difficulties of lutein due to its poor solubility in water, poor stability and low oral absorption rate, the purpose of the present invention is to disclose a method for preparing a lutein compound preparation with good water solubility, strong stability and high absorption rate through the innovative joint use of synergistic absorption-promoting nutrient compatibility technology, pulse electric field coupled high-voltage homogenization technology and low-temperature ion-driven self-assembly technology, and to use it for the production of vision protection functional foods. The processing method has high embedding rate, good safety and low production cost, and the lutein compound preparation product has strong stability and high absorption rate, and the vision protection functional food developed with the lutein compound preparation as the core functional ingredient has good efficacy.

[0004] To achieve the above object, the present invention provides a highly absorbable lutein composite preparation, characterized in that the highly absorbable lutein composite preparation is a water-soluble particle formed by molecular self-assembly of lutein, dipotassium glycyrrhizinate, triterpenic acid, protein, and dietary polysaccharide, wherein the mass ratio of lutein, dipotassium glycyrrhizinate, and triterpenic acid is 1:10:5 to 1:50:1.

[0005] Furthermore, the highly absorbable lutein compound preparation is in the form of capsules, tablets, granules, oral liquid or beverage.

[0006] The present invention also provides a method for preparing the highly absorbable lutein composite preparation, which is characterized by comprising the following steps:

[0007] S1. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles: After dissolving lutein, dipotassium glycyrrhizinate and triterpenic acid (solvents may be used to assist dissolution, such as anhydrous ethanol or a menthol-thymol mixture (molar ratio of 1:1)), stirring evenly and ultrasonicating to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles; the ultrasonication here not only has the effect of mixing, but also can promote the self-assembly of these components into smaller aggregates, which is conducive to the subsequent protein encapsulation;

[0008] Preferably, the mass ratio of lutein, dipotassium glycyrrhizinate and triterpene acid is 1:10:5 to 1:50:1, and the ultrasonic time is 2±0.5 min;

[0009] S2. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex: Add lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles to the protein solution treated by the electric field, preferably, the mass ratio of lutein to protein is 1: (25 ± 5); mix; homogenize the mixed solution with a high-pressure homogenizer to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution with protein as the shell and lutein, dipotassium glycyrrhizinate and triterpenic acid as the core; denature the protein solution, there are many methods, such as pH adjustment, acid-base reagents will be used; ultrasound can also be used, but it is not stable enough, and the heat generation is large, which will destroy the thermosensitive active ingredients. Comprehensive comparison, the pulsed electric field has the advantages of being green, efficient and stable. The protein treated by the electric field will undergo sufficient partial denaturation, thereby promoting the self-assembly of the partially denatured protein with aggregates such as saponin and dipotassium glycyrrhizinate, and adsorbing on the surface of the aggregate to achieve the purpose of encapsulation. The high-pressure homogenization can break up the self-assembled aggregates and reduce their particle size, thereby improving the solubility of the system and the stability in aqueous solution.

[0010] Preferably, the electric field treated protein solution is a 2±0.5 mass % protein aqueous solution treated with a pulsed electric field, so that the protein solution changes from a clear state to a turbid state, and the protein is partially denatured;

[0011] S3. Preparation of lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex: first add antifreeze peptide and then dietary polysaccharide to the lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein complex aqueous solution; mix well and place the mixed solution in a cold storage; when the temperature of the mixed solution drops to a low temperature below zero, adjust the pH of the system to acidic to allow the polysaccharide to adsorb to the surface of the protein complex, thereby obtaining a lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex suspension;

[0012] Preferably, the concentration of antifreeze peptide added is 1±0.3 mass %; the final concentration of dietary polysaccharide added is 0.5±0.2 mass %; the mixture is mixed and the acidic pH is 4.0 at a low temperature of -2°C below zero.

[0013] Furthermore, the method also includes centrifugation and freeze-drying to obtain a highly absorbable lutein compound preparation dry powder.

[0014] Furthermore, the protein is at least one of bovine serum albumin, ovalbumin, whey protein, peanut protein and soy protein.

[0015] Furthermore, the dietary polysaccharide is at least one of pectin, hyaluronic acid, fucoidan and galactan sulfate.

[0016] Furthermore, in the step S2, the intensity of the pulse electric field is 30±5 kV / cm, and the treatment time is 6 to 8 minutes.

[0017] Furthermore, in the step S2, the pressure of high-pressure homogenization is 60 MPa to 120 MPa, and the homogenization time is 1 to 5 min.

[0018] Furthermore, in the step S3, the molecular weight of the antifreeze peptide is 500Da to 2000Da.

[0019] The present invention also protects the use of the highly absorbable water-soluble lutein compound preparation or the highly absorbable water-soluble lutein compound preparation prepared by the preparation method in vision-protecting functional foods.

[0020] The embedding rate of lutein in the preparation step (3) of the lutein composite preparation is ≥98%. The particle size of the lutein composite preparation is 500-800 nm. The lutein composite preparation is heated at 121° C. for 15 minutes, and the lutein retention rate is ≥90%, ensuring that the lutein will not be degraded in the processing environment such as sterilization, thereby ensuring the functional quality of the product. The oral absorption efficiency of lutein in the highly absorbable lutein composite preparation is increased by 20-35 times.

[0021] The present invention innovatively adopts the nutritional compatibility technology to promote intestinal absorption. By screening and matching dietary ingredients that can upregulate the expression of transmembrane transport proteins responsible for lutein and help lutein form digestive micelles, the efficiency of lutein transport across the intestinal epithelium is effectively improved. At the same time, pulse electric field coupled high-voltage homogenization technology and low-temperature ion-driven self-assembly technology are used to physically encapsulate lutein and its compatible ingredients, and simultaneously improve its water solubility, processing stability and digestion stability, solving its application problems in the food industry. Therefore, the highly absorbable lutein composite preparation prepared by the present invention has the characteristics of good absorption effect, easy water solubility and high stability. The vision protection functional food prepared with this raw material as the core functional ingredient has good effect, is green and safe.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] The present invention innovatively combines absorption-promoting nutrient compatibility technology, pulse electric field coupled high-voltage homogenization technology, and low-temperature ion-driven self-assembly technology to prepare a water-soluble lutein composite preparation with an outer shell of protein and dietary polysaccharides and an inner core of lutein, dipotassium glycyrrhizinate, and triterpene acid. Compared with the prior art, the beneficial effects of the present invention are mainly reflected in:

[0024] (1) High encapsulation efficiency. In view of the fact that lutein is sensitive to acid and alkali and easily decomposed by heat, during the encapsulation process of lutein, pulse electric field coupled high-voltage homogenization technology is used, without the use of heating and acid-base reagents, so that the processing environment of protein encapsulated lutein is always at a low temperature and neutral pH; at the same time, low-temperature ion-driven self-assembly technology is used to further modify the surface molecules of protein particles, and low temperature conditions (such as -2°C) are used during the processing process, thereby effectively reducing the damage to lutein caused by processing conditions, and the encapsulation rate of lutein reaches more than 98%.

[0025] (2) Strong oral absorption effect. In view of the fact that lutein is closely regulated by lipid transport proteins during transmembrane transport, the present invention innovatively adopts absorption-promoting nutritional combination technology, selects dipotassium glycyrrhizinate and triterpene acids from a variety of dietary ingredients for combination with lutein, and effectively improves the intestinal absorption efficiency of lutein by regulating the expression of lipid transport proteins responsible for the absorption of lutein across the intestinal epithelium, thereby increasing the oral absorption efficiency of lutein by 20 to 35 times.

[0026] (3) The product has good vision protection effect. By greatly improving the oral absorption rate of lutein, the physiological and nutritional functions of lutein in vision protection are effectively improved, and the developed product has good vision protection effect. DETAILED DESCRIPTION

[0027] Embodiments of the present invention are described in detail below, and the examples of the embodiments are intended to be used to explain the present invention, but should not be construed as limiting the present invention. In the embodiments, if specific techniques or conditions are not specified, they are carried out according to the techniques or conditions described in the literature in this area or according to the product specification. If the manufacturer is not specified for reagents or instruments used, they are all conventional products that can be obtained commercially.

[0028] A highly absorbable lutein composite preparation, characterized in that the highly absorbable lutein composite preparation is a water-soluble particle formed by molecular self-assembly of lutein, dipotassium glycyrrhizinate, triterpenic acid, protein, and dietary polysaccharide, wherein the mass ratio of lutein, dipotassium glycyrrhizinate, and triterpenic acid is 1:10:5 to 1:50:1.

[0029] Furthermore, the highly absorbable lutein compound preparation is in the form of capsules, tablets, granules, oral liquid or beverage.

[0030] The present invention also provides a method for preparing the highly absorbable lutein composite preparation, which is characterized by comprising the following steps:

[0031] S1. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles: After dissolving lutein, dipotassium glycyrrhizinate and triterpenic acid (solvents may be used to assist dissolution, such as anhydrous ethanol or a menthol-thymol mixture (molar ratio of 1:1)), stirring evenly and ultrasonicating to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles; the ultrasonication here not only has the effect of mixing, but also can promote the self-assembly of these components into smaller aggregates, which is conducive to the subsequent protein encapsulation;

[0032] Preferably, the mass ratio of lutein, dipotassium glycyrrhizinate and triterpene acid is 1:10:5 to 1:50:1, and the ultrasonic time is 2±0.5 min;

[0033] S2. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex: Add lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles to the protein solution treated by the electric field, preferably, the mass ratio of lutein to protein is 1: (25 ± 5); mix; homogenize the mixed solution with a high-pressure homogenizer to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution with protein as the shell and lutein, dipotassium glycyrrhizinate and triterpenic acid as the core; denature the protein solution, there are many methods, such as pH adjustment, acid-base reagents will be used; ultrasound can also be used, but it is not stable enough, and the heat generation is large, which will destroy the thermosensitive active ingredients. Comprehensive comparison, the pulsed electric field has the advantages of being green, efficient and stable. The protein treated by the electric field will undergo sufficient partial denaturation, thereby promoting the self-assembly of the partially denatured protein with aggregates such as saponin and dipotassium glycyrrhizinate, and adsorbing on the surface of the aggregate to achieve the purpose of encapsulation. The high-pressure homogenization can break up the self-assembled aggregates and reduce their particle size, thereby improving the solubility of the system and the stability in aqueous solution.

[0034] Preferably, the electric field treated protein solution is a 2±0.5 mass % protein aqueous solution treated with a pulsed electric field, so that the protein solution changes from a clear state to a turbid state, and the protein is partially denatured;

[0035] S3. Preparation of lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex: first add antifreeze peptide and then dietary polysaccharide to the lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein complex aqueous solution; mix well and place the mixed solution in a cold storage; when the temperature of the mixed solution drops to a low temperature below zero, adjust the pH of the system to acidic to allow the polysaccharide to adsorb to the surface of the protein complex, thereby obtaining a lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex suspension;

[0036] Preferably, the concentration of antifreeze peptide added is 1±0.3 mass %; the final concentration of dietary polysaccharide added is 0.5±0.2 mass %; the mixture is mixed and the acidic pH is 4.0 at a low temperature of -2°C below zero.

[0037] Furthermore, the method also includes centrifugation and freeze-drying to obtain a highly absorbable lutein compound preparation dry powder.

[0038] Furthermore, the protein is at least one of bovine serum albumin, ovalbumin, whey protein, peanut protein and soy protein.

[0039] Furthermore, the dietary polysaccharide is at least one of pectin, hyaluronic acid, fucoidan and galactan sulfate.

[0040] Furthermore, in the step S2, the intensity of the pulse electric field is 30±5 kV / cm, and the treatment time is 6 to 8 minutes.

[0041] Furthermore, in the step S2, the pressure of high-pressure homogenization is 60 MPa to 120 MPa, and the homogenization time is 1 to 5 min.

[0042] Furthermore, in the step S3, the molecular weight of the antifreeze peptide is 500Da to 2000Da.

[0043] Example 1

[0044] Preparation of lutein compound preparation:

[0045] S1. Lutein, dipotassium glycyrrhizinate, and triterpenic acid were weighed in a mass ratio of 1:10:5, dissolved with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles;

[0046] S2. The bovine serum albumin was dissolved in deionized water, and the protein concentration was 1.5 mass %; the protein aqueous solution was treated with a pulsed electric field of 35 kV / cm for 6 min, and then lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles were added to make the mass ratio of lutein to protein 1:20, and mixed with a stirrer; the mixed solution was homogenized at a pressure of 60 MPa for 5 min using a high-pressure homogenizer to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution;

[0047] S3. Add antifreeze peptide with an average molecular weight of 500Da to a concentration of 0.7% by mass; then add pectin to a final concentration of 0.3% by mass, mix well and place in a -2°C cold storage; when the temperature of the mixed solution drops to -2°C, adjust the pH of the system to 4.0 to allow pectin to adsorb to the surface of the protein complex, and obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein-polysaccharide complex suspension, with a lutein encapsulation efficiency of 98.3% and a particle size of 500nm;

[0048] S4. Centrifuge and freeze-dry to obtain a lutein compound preparation dry powder. The lutein compound preparation dry powder was redissolved in deionized water and heated at 121° C. for 15 minutes. The retention rate of lutein was 91.8%.

[0049] Preparation of vision protection functional food: Mix the lutein compound preparation with appropriate amount of magnesium stearate and starch, sieve and fill into hard capsules using a fully automatic capsule filling machine. The filling amount of each capsule is 0.5g, and each capsule contains 40mg of lutein, so as to obtain capsule functional food.

[0050] Example 2

[0051] Preparation of lutein compound preparation:

[0052] S1. Lutein, dipotassium glycyrrhizinate, and triterpenic acid were weighed in a mass ratio of 1:25:3, dissolved with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles;

[0053] S2. The egg white albumin was dissolved in deionized water, and the protein concentration was 2 mass %; the protein aqueous solution was treated with a pulsed electric field with an intensity of 30 kV / cm for 7 minutes, and then lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles were added to make the mass ratio of lutein to protein 1:25, and mixed with a stirrer; the mixed solution was homogenized with a high-pressure homogenizer at a pressure of 80 MPa for 4 minutes to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution;

[0054] S3. Add antifreeze peptide with an average molecular weight of 1000Da to a concentration of 1% by mass; then add fucoidan to a final concentration of 0.5% by mass, mix well and place in a -2°C cold storage; when the temperature of the mixed solution drops to -2°C, adjust the pH of the system to 4.0 to allow fucoidan to adsorb to the surface of the protein complex, and obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein-polysaccharide complex suspension, with a lutein encapsulation efficiency of 98.5% and a particle size of 700nm;

[0055] S4. Centrifuge and freeze-dry to obtain a lutein compound preparation dry powder. The lutein compound preparation dry powder was redissolved in deionized water and heated at 121° C. for 15 minutes. The retention rate of lutein was 90.0%.

[0056] Preparation of vision protection functional food: Mix the lutein compound preparation with an appropriate amount of one or more auxiliary materials selected from dextrin, starch, microcrystalline cellulose and lactose, moisten with an appropriate amount of 70% (v / v) ethanol, make a soft material, granulate through a 30-mesh sieve, dry at 70-80°C, sieve the whole granules through a 60-mesh sieve, and press into tablets, each tablet weighing 0.25g and containing 20mg lutein, thus obtaining a tablet-type functional food.

[0057] Example 3

[0058] Preparation of lutein compound preparation:

[0059] S1. Lutein, dipotassium glycyrrhizinate, and triterpenic acid were weighed in a mass ratio of 1:40:2.5, dissolved with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles;

[0060] S2. The peanut protein powder was dissolved in deionized water, and the protein concentration was 2.5 mass %; the protein aqueous solution was treated with a pulsed electric field with an intensity of 25 kV / cm for 8 minutes, and then lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles were added to make the mass ratio of lutein to protein 1:30, and mixed with a stirrer; the mixed solution was homogenized with a high-pressure homogenizer at a pressure of 100 MPa for 2 minutes to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution;

[0061] S3. Add antifreeze peptide with an average molecular weight of 1500Da to a concentration of 1.3% by mass; then add sulfated galactan to a final concentration of 0.7% by mass, mix well and place in a -2°C cold storage; when the temperature of the mixed solution drops to -2°C, adjust the system pH to 4.0 to allow the sulfated galactan to adsorb onto the surface of the protein complex, and obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein-polysaccharide complex suspension, with a lutein encapsulation efficiency of 98.1% and a particle size of 800nm;

[0062] S4. Centrifuge and freeze-dry to obtain a lutein compound preparation dry powder. The lutein compound preparation dry powder was redissolved in deionized water and heated at 121° C. for 15 minutes. The retention rate of lutein was 93.7%.

[0063] Preparation of vision protection functional food: mix the lutein compound preparation with appropriate amount of one or more of soluble starch, dextrin, sucrose, lactose and mannitol, moisten with appropriate amount of 70% (v / v) ethanol, make a soft material, granulate through a 20-mesh sieve, dry at 70-80°C, and granulate with 60 mesh. The lutein content is 80mg / 12g (one small bag), and the functional food of granule type is obtained.

[0064] Example 4

[0065] Preparation of lutein compound preparation:

[0066] S1. Lutein, dipotassium glycyrrhizinate, and triterpenic acid were weighed in a mass ratio of 1:50:1, dissolved with a small amount of anhydrous ethanol, stirred evenly, and ultrasonicated for 2 minutes to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles;

[0067] S2. Dissolve soy protein powder in deionized water, the protein concentration is 2 mass %; the protein aqueous solution is treated with a pulsed electric field with an intensity of 30 kV / cm for 7 min, and then add lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles to make the mass ratio of lutein to protein 1:25, and mix with a stirrer; the mixed solution is homogenized with a high-pressure homogenizer at a pressure of 120 MPa for 1 min to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution;

[0068] S3. Add antifreeze peptide with an average molecular weight of 2000Da to a concentration of 1% by mass; then add hyaluronic acid to a final concentration of 0.5% by mass, mix well and place in a -2°C cold storage; when the temperature of the mixed solution drops to -2°C, adjust the pH of the system to 4.0 to allow hyaluronic acid to adsorb to the surface of the protein complex, and obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein-polysaccharide complex suspension, with a lutein encapsulation efficiency of 98.7% and a particle size of 720nm;

[0069] S4. Centrifuge and freeze-dry to obtain a lutein compound preparation dry powder. The lutein compound preparation dry powder was redissolved in deionized water and heated at 121° C. for 15 minutes. The retention rate of lutein was 92.5%.

[0070] Preparation of vision protection functional food: dissolve the lutein compound preparation in water with a lutein concentration of 40 mg / 100 mL, add appropriate amount of sweetener and stabilizer, mix well, homogenize, filter, fill, and sterilize to obtain a beverage-type functional food.

[0071] Example 5 (Evaluation of absorption effect)

[0072] Test samples:

[0073] Table 1 Absorption effect of tested samples

[0074]

[0075]

[0076] Experimental animals: SPF grade male ICR mice, 15-20 g, purchased from Shanghai Slake Experimental Animal Co., Ltd. Basic feed ingredients for mice: carbohydrate 64%, protein 21%, fat 4%, fiber 5%, water 6%. The above feeds were purchased from Shanghai Slake Experimental Animal Co., Ltd. The temperature of the feeding room was 23±2℃, and the relative humidity was 55±5%.

[0077] Animal grouping and treatment method: After 1 week of adaptive feeding, the mice were randomly divided into 20 groups according to body weight, with 7 mice in each group, including a blank group (gavage with deionized water) and the above 23 groups of test samples (gavage with test solution). The above 23 groups were dissolved in deionized water and used as the test solution. Each mouse was gavaged with 200 μL of test solution or deionized water each time, and the dose of lutein was 10 mg / kg. After 4 hours of gavage, the mouse eyeballs were removed to collect blood, and the changes in the content of lutein in plasma were analyzed by high performance liquid chromatography (HPLC). The content of lutein in plasma was calculated according to the standard curve method to reflect the intestinal absorption efficiency of lutein. The small intestine was taken and homogenized, and the total protein was extracted. The expression of lipid transport proteins SR-B1 and CD36 in the small intestine was analyzed by Elisa kit.

[0078] Experimental results:

[0079] Table 2. Lutein absorption effect evaluation table

[0080]

[0081]

[0082] HPLC analysis of lutein concentration in the serum of mice in each treatment group is shown in Table 2. The results show that the serum concentration of unencapsulated lutein after oral gavage was only 24.39 pmol / mL; when lutein was combined with dipotassium glycyrrhizinate, it showed a good absorption-promoting effect in the mass ratio range of 1:10 to 1:50, but the absorption effect of lutein was not further improved by further increasing dipotassium glycyrrhizinate. For example, when the mass ratio of lutein to dipotassium glycyrrhizinate was further increased to 1:60 or 1:70, there was no significant change in the absorption effect of lutein compared with 1:50 (P<0.05), indicating that the improvement of the absorption effect of lutein by increasing dipotassium glycyrrhizinate was ineffective. Correspondingly, the expression of SR-B1 and CD36, the transport proteins responsible for lutein across the intestinal epithelium, also increased accordingly, indicating that dipotassium glycyrrhizinate can promote the intestinal absorption of lutein by upregulating the expression of transport proteins. Further introduction of triterpenic acid revealed that when the ratio of dipotassium glycyrrhizinate was low, more triterpenic acid needed to be introduced. For example, when the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid was 1:10:5, the serum concentration of lutein reached 415.67 pmol / mL, an increase of 16.04 times. When the ratio of dipotassium glycyrrhizinate was high, relatively less triterpenic acid could be introduced. For example, when the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid was 1:50:1, the serum concentration of lutein reached 818.57 pmol / mL, an increase of 32.5 6 times; if the proportion of triterpenic acid is increased, when the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid is 1:50:3 or the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid is 1:50:5, although the serum concentration of lutein can reach 821.46±40.92pmol / mL and 823.49±39.34pmol / mL respectively, there is no significant difference compared with 818.57±32.67pmol / mL when the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid is 1:50:1 (P<0.05). This shows that when the proportion of dipotassium glycyrrhizinate is high, excessive introduction of triterpenic acid is not beneficial to improving the intestinal absorption of lutein. Therefore, the mass ratio of lutein, dipotassium glycyrrhizinate, and triterpenic acid is 1:10:5-1:50:1, which has a better absorption-promoting effect.

[0083] On this basis, the method of the present invention was used to molecularly embed lutein, dipotassium glycyrrhizinate and triterpenic acid, and the absorption effect of the lutein composite preparations of Examples 1 to 4 was evaluated. The results showed that when protein was used to encapsulate lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles, taking Example 1 as an example (the mass ratio of lutein + dipotassium glycyrrhizinate + triterpenic acid was 1:10:5), when the mass ratio of lutein to protein was 1:15 (Group 16: lutein serum concentration 414.76±17.36pmol / mL), compared with the effect without protein encapsulation (Group 12: lutein serum concentration 415.67±21.35pmol / mL), the absorption effect of lutein was not significantly improved (P<0.05); however, when the mass ratio of protein was further increased, the mass ratio of lutein to protein was increased to 1:20 and 1:30 (Groups 17 and 18), the serum concentration of lutein was increased to 474.13±18.14pmol / mL and 512.24±19.13pmol / mL, respectively, which had a significant improvement effect (P<0.05). However, when the protein concentration is further increased, the mass of lutein to bovine serum albumin reaches 1:35, and the serum concentration of lutein is no different from that at 1:30. This shows that the ratio of lutein to protein is in a reasonable range of 1:20-30. Further adsorption of polysaccharides on the protein shell also has an absorption-promoting effect. The difference between Example 1 (Group 20) and Group 17 is that Group 17 adsorbs pectin on the outside of the protein shell, thereby increasing the serum concentration of lutein from 474.13±18.14pmol / mL to 517.91±19.31pmol / mL, an increase of 9.23%, and has good thermal stability. After heating at 121°C for 15 minutes, the retention rate of lutein is 91.8%. After the lutein composite preparations of Examples 2 to 4 were gavaged into mice, their serum concentrations reached 682.92 pmol / mL, 756.09 pmol / mL, and 878.53 pmol / mL, respectively. Compared with the unencapsulated lutein, the absorption effect was increased by 27 times, 30 times, and 35.02 times, respectively. At the same time, the expression of the transport proteins SR-B1 and CD36 responsible for the absorption of lutein across the intestinal epithelium was also increased accordingly. This shows that the absorption-promoting nutrient compatibility technology, pulsed electric field coupled high-voltage homogenization technology, and low-temperature ion-driven self-assembly technology jointly adopted by the present invention can achieve efficient oral absorption of lutein by regulating the expression of intestinal epithelial transport proteins and improving the stability of lutein.

[0084] Example 6 (Evaluation of Vision Protection Efficacy)

[0085] Test samples: the samples obtained by grouping as 1, 4, 7, 12, 13, 20-24 in Table 1.

[0086] Sample treatment: Dissolve the formulated product directly in deionized water at a mass-to-volume ratio of 1:10 (if necessary, a co-solvent can be used to assist dissolution, such as anhydrous ethanol or a menthol-thymol mixture (molar ratio 1:1)), which is the test solution.

[0087] Experimental animals and feeding environment: SPF-grade blue rabbits, half male and half female, weighing 2.0±0.2kg, were provided by the Animal Center of Beijing Tiantan Biological Products Co., Ltd. The indoor ventilation conditions were good, with normal day and night changes (8:00am-8:00pm), relative humidity of 55±5%, and room temperature: 23±2℃.

[0088] Animal grouping and treatment methods: The blue rabbits were adaptively fed for one week and fed freely. After one week, they were randomly divided into 11 groups (7 experimental animals in each group) according to body weight, namely the normal group, the model group and 9 intervention groups (the above test samples). Feeding continued for 2 weeks at the following doses: the normal group was gavaged with 2.5 mL of water per day, the model group was gavaged with 2.5 mL of water per day, and each intervention group was gavaged with lutein at a dose of 50 mg / kg / d. After 2 weeks, a light experiment was conducted. A homemade light box was used for the light experiment: the light box had a size of 50×40×30 cm, with mirrors on the bottom and sides, and a white cold light source fluorescent tube was installed on each of the four corners of the side, so that the light intensity in all directions in the light box was 15000±1000 lux (measured by TES-1332A illuminometer). The normal group continued to be fed conventionally. After the blue rabbits in the model group and the intervention group were dark-adapted in a 60-100 lux light environment for 24 hours, the compound tropicamide eye drops were used to dilate the pupils of both eyes. After 20 minutes, they were placed in a homemade light box for illumination. A blue rabbit was placed in each light box for 2 hours, and the activities of the blue rabbits were not restricted. The ventilation was maintained during the entire illumination process to ensure that the temperature in the light box was maintained at a relatively constant temperature of 21-24°C. After the illumination treatment, the experimental animals were returned to the animal room for dark adaptation. The animals in each group were continued to be fed for 7 days according to the aforementioned gavage dose and feeding method, and then the kit method was used to analyze the changes in the total antioxidant capacity of the eyes. The maximum mixed reaction ERG value of the retina of each group of animals was detected by the electro-visual physiology ERG to evaluate the changes in retinal function.

[0089] Experimental results:

[0090] Table 3 Vision protection efficacy evaluation table

[0091]

[0092]

[0093] The results are shown in Table 3. Excessive light irradiation has a great damage to the eyes. The total antioxidant capacity of the eyes and the maximum mixed reaction ERG of the retina decreased by 62.8% and 58.0%, respectively. When lutein is used alone for intervention, it can have a certain vision protection function, but the effect is not ideal. Compared with the model group, the total antioxidant capacity of the eyes and the maximum mixed reaction ERG of the retina only increased by 18.3% and 19.4%, respectively. When lutein is combined with dipotassium glycyrrhizinate to improve the oral absorption efficiency of lutein (as shown in Table 2), its vision protection function is further improved. Further combined with triterpene acid to further improve the oral absorption efficiency of lutein (as shown in Table 2), the vision protection function of lutein is further improved. For example, when the mass ratio of lutein to dipotassium glycyrrhizinate and monoterpene acid is 1:50:1, compared with the model group, the total antioxidant capacity of the eyes and the maximum mixed reaction ERG of the retina increased by 1.2 times and 1.0 times, respectively. Further intervention evaluation was performed on the lutein composite preparations of Examples 1 to 4 of the present invention, and all showed good eye protection function. When the mass ratio of lutein to dipotassium glycyrrhizinate and monoterpene acid was 1:50:1, when not embedded (i.e., Group 13), the total antioxidant capacity of the eyes was 1.59 ± 0.02 mmol / g protein, and the maximum mixed reaction ERG of the retina was 249.34 ± 6.85 mV. When embedded (i.e., Group 23), the total antioxidant capacity of the eyes was 1.78 ± 0.03 mmol / g protein, and the maximum mixed reaction ERG of the retina was 279.34 ± 6.51 mV. It can be seen that even if the dosage of the three is the same, the total antioxidant capacity of the eyes and the maximum mixed reaction ERG of the retina of the product using the process of the present invention are significantly improved.

[0094] When the mass ratio of lutein to dipotassium glycyrrhizinate and monoterpene acid was 1:10:5, when not embedded (i.e., group 12), the total antioxidant capacity of the eyes was 1.38±0.03mmol / g protein, and the maximum mixed reaction ERG of the retina was 207.52±6.21mV. When embedded (i.e., group 20), the total antioxidant capacity of the eyes was 1.41±0.03mmol / g protein, and the maximum mixed reaction ERG of the retina was 217.42±7.66mV. It can be seen that even if the dosage of the three is the same, the total antioxidant capacity of the eyes and the maximum mixed reaction ERG of the retina of the product using the process of the present invention are significantly improved.

[0095] Combined with the results of promoting lutein absorption in Table 2, it can be seen that the effect of promoting oral absorption of lutein is significantly positively correlated with its eye protection function.

[0096] The above results indicate that the vision-protecting functional food disclosed in the present invention can effectively prevent vision damage caused by excessive light irradiation, increase the total antioxidant capacity of the eyes and improve the function of the retina.

[0097] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention without departing from the principles and intent of the present invention.

Claims

1. A highly absorbable lutein composite preparation, characterized in that: The highly absorbable lutein compound preparation is a water-soluble particle formed by molecular self-assembly of lutein, dipotassium glycyrrhizinate, triterpenic acid, protein, and dietary polysaccharide, wherein the mass ratio of lutein, dipotassium glycyrrhizinate, and triterpenic acid is 1:10:5 to 1:50:

1.

2. The highly absorbable lutein composite preparation according to claim 1, characterized in that: The highly absorbable lutein compound preparation is in the form of capsules, tablets, granules, oral liquid or beverage.

3. The method for preparing the highly absorbable lutein composite preparation according to claim 1, characterized in that: The steps include: S1. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles: Dissolve lutein, dipotassium glycyrrhizinate and triterpenic acid, stir evenly, and perform ultrasound to obtain lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles; Preferably, the mass ratio of lutein, dipotassium glycyrrhizinate and triterpene acid is 1:10:5 to 1:50:1, and the ultrasonic time is 2±0.5 min; S2. Preparation of lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex: adding lutein-dipotassium glycyrrhizinate-triterpenic acid fusion micelles to the protein solution treated by the electric field, preferably, the mass ratio of lutein to protein is 1:(25±5); mixing; homogenizing the mixed solution with a high-pressure homogenizer to obtain a lutein-dipotassium glycyrrhizinate-triterpenic acid-protein complex aqueous solution with protein as the shell and lutein, dipotassium glycyrrhizinate and triterpenic acid as the core; Preferably, the electric field treated protein solution is a 2±0.5 mass% protein aqueous solution treated with a pulsed electric field, so that the protein solution changes from a clear state to a turbid state, and the protein is partially denatured; S3. Preparation of lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex: first add antifreeze peptide and then dietary polysaccharide to the lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein complex aqueous solution; mix well and place the mixed solution in a cold storage; when the temperature of the mixed solution drops to a low temperature below zero, adjust the pH of the system to acidic to allow the polysaccharide to adsorb to the surface of the protein complex, thereby obtaining a lutein-dipotassium glycyrrhizinate-triterpenoid acid-protein-polysaccharide complex suspension; Preferably, the concentration after adding the antifreeze peptide is 1±0.3 mass %; the final concentration after adding the dietary polysaccharide is 0.5±0.2 mass %; Mix well, and at a sub-zero temperature of -2°C, the acidic pH is 4.

0.

4. The method for preparing the highly absorbable lutein composite preparation as claimed in claim 3, characterized in that: The method also includes centrifugation and freeze-drying to obtain a highly absorbable lutein compound preparation dry powder.

5. The method for preparing the highly absorbable lutein composite preparation as claimed in claim 3, characterized in that: The protein is at least one of bovine serum albumin, ovalbumin, whey protein, peanut protein and soy protein.

6. The method for preparing the highly absorbable lutein composite preparation as claimed in claim 3, characterized in that: The dietary polysaccharide is at least one of pectin, hyaluronic acid, fucoidan and galactan sulfate.

7. The method for preparing the highly absorbable lutein composite preparation as claimed in claim 3, characterized in that: In the step S2, the intensity of the pulse electric field is 30±5 kV / cm, and the treatment time is 6 to 8 minutes.

8. The method for preparing the highly absorbable lutein composite preparation as claimed in claim 3, characterized in that: In the step S2, the pressure of high-pressure homogenization is 60 MPa to 120 MPa, and the homogenization time is 1 to 5 minutes.

9. The method for preparing the highly absorbable lutein composite preparation according to claim 3, characterized in that: In the step S3, the molecular weight of the antifreeze peptide is 500Da to 2000Da.

10. Use of the highly absorbable water-soluble lutein compound preparation according to any one of claims 1-2 or the highly absorbable water-soluble lutein compound preparation prepared by the preparation method according to any one of claims 2-9 in vision-protecting functional foods.

Images