Polypeptide composition for regulating menstruation and nourishing blood and preparation method thereof

Abstract

The invention relates to the technical field of functional food, in particular to a polypeptide composition for regulating menstruation and nourishing blood and a preparation method thereof, and the polypeptide composition comprises the following raw materials: a Chinese wolfberry extract, an astaxanthin-fish oil co-coated preparation, a red algae oligosaccharide iron complex, maltodextrin and folic acid. In the polypeptide composition for regulating menstruation and nourishing blood and the preparation method thereof, the astaxanthin-fish oil co-coated preparation stably wraps the strong antioxidant astaxanthin and the fish oil through a complex coacervation microencapsulation technology, so that excessive free radicals generated by menstrual physiological stress can be efficiently eliminated, inflammatory response of uterine tissues can be relieved, and discomfort symptoms such as dysmenorrhea and the like can be relieved; according to the red algae oligosaccharide iron complex, oligosaccharide is used as a natural carrier of an iron element and is absorbed in an oligosaccharide iron chelating form in intestinal tracts, the absorption way of traditional ferric salt is avoided, the bioavailability is greatly improved, the iron element can be efficiently and mildly supplemented, and iron-deficiency anemia is improved.

Description

Technical Field

[0001] This invention relates to the field of functional food technology, and more specifically, to a polypeptide composition for regulating menstruation and nourishing blood, and its preparation method. Background Technology

[0002] Menstrual irregularities, anemia, and related discomforts are common health problems that plague many women. Currently, while traditional Chinese medicine formulas (such as Siwu Tang) have certain therapeutic effects, they are slow to take effect, have poor taste, and are inconvenient to carry. Modern nutritional supplements (such as iron supplements and vitamins) have limited functions, and iron supplements are prone to causing gastrointestinal irritation and metallic taste, have low bioavailability, and provide a poor user experience. In recent years, with the development of nutrition science and food engineering, compound functional foods have gradually become a new trend.

[0003] However, the active ingredients (such as unsaturated fatty acids and carotenoids) are chemically unstable and are prone to oxidative degradation during processing and storage. Furthermore, the physical properties (such as oil solubility / water solubility) of different ingredients vary greatly, making it difficult to ensure the uniformity and stability of the product through simple mixing, and thus difficult to achieve synergistic effects and targeted delivery. In view of this, we propose a peptide composition for regulating menstruation and nourishing blood, as well as its preparation method. Summary of the Invention

[0004] The purpose of this invention is to provide a polypeptide composition for regulating menstruation and nourishing blood, and its preparation method, in order to solve the problems mentioned in the background art, such as the unstable chemical properties of active ingredients (e.g., unsaturated fatty acids, carotenoids), which are prone to oxidative degradation during processing and storage, and the large differences in the physical properties (e.g., oil solubility / water solubility) of different components, making it difficult to ensure the uniformity and stability of the product through simple mixing, and making it difficult to achieve synergistic effects and targeted delivery.

[0005] This invention provides a polypeptide composition for regulating menstruation and nourishing blood, comprising the following raw materials: wolfberry extract, astaxanthin-fish oil co-encapsulation preparation, red algae oligosaccharide iron complex, maltodextrin and folic acid; The astaxanthin-fish oil co-encapsulated formulation is prepared by combining an active oil phase with phospholipids, emulsifying the emulsion through a hydrophobic ceramic membrane, and then encapsulating it using a gelatin-gum arabic composite coagulation method.

[0006] Preferably, the ingredients are: 5-15 parts by weight of wolfberry extract, 10-20 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 15-25 parts by weight of red algae oligosaccharide iron complex, 30-50 parts by weight of maltodextrin, and 0.5-1.5 parts by weight of folic acid.

[0007] Preferably, the preparation method of the astaxanthin-fish oil co-encapsulated formulation is as follows: Astaxanthin oil extract and fish oil were mixed at a mass ratio of 1:10-15, and vitamin E and rosemary extract were added. The mixture was stirred at 50-100 rpm under nitrogen protection at 30-40°C to obtain the oil phase. The oil phase and lecithin were mixed at a mass ratio of 1:1-1.5 and stirred at 100-200 rpm for 1-2 hours under nitrogen protection at 50-60℃ to obtain the phospholipid complex oil phase. Dissolve 1-2% of Tween 80 (by mass of the phospholipid complex oil phase) in deionized water and preheat to 50-55°C to obtain a continuous phase. Extrude the phospholipid complex oil phase into the continuous phase through a hydrophobic ceramic membrane with a pore size of 0.8-1.2 μm under a pressure of 0.1-0.3 MPa to obtain the primary emulsion. The colostrum was added to the composite wall material solution while stirring at 100-200 rpm, with the mass ratio of the phospholipid complex oil phase to the composite wall material being 1:1.0-2.0. The temperature was maintained at 40-45℃, and 0.1 mol / L citric acid solution was added dropwise to adjust the pH to 4.0-4.5, and the mixture was stirred for 30-45 min. The temperature was lowered to 5-10℃, and 0.05-0.1% of transglutaminase by the mass of the colostrum was added for cross-linking for 2-4 h. After the reaction was completed, the wet microcapsules were collected through a 200-mesh sieve and washed 2-3 times with deionized water. The wet microcapsules were redispersed in deionized water with a solid content of 20%-30% to form a suspension. The suspension was sprayed into liquid nitrogen to obtain frozen particles; the frozen particles were dried at temperatures below -40°C and below 10 Pa to obtain an astaxanthin-fish oil co-encapsulated formulation.

[0008] Preferably, the amount of vitamin E added is 0.1-0.3% of the total mass of the oil phase, and the amount of rosemary extract added is 0.05-0.1% of the total mass of the oil phase.

[0009] Preferably, the composite wall material solution is obtained by mixing a 3% gelatin solution and a 3% gum arabic solution at a mass ratio of 1:1.

[0010] On the other hand, the present invention provides a method for preparing a menstrual regulating and blood-nourishing polypeptide composition, which includes the following steps: S1.1 Weigh the following raw materials in parts by weight: 5-15 parts by weight of wolfberry extract, 10-20 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 15-25 parts by weight of red algae oligosaccharide iron complex, 30-50 parts by weight of maltodextrin and 0.5-1.5 parts by weight of folic acid. S1.2 Add wolfberry extract to deionized water at a volume ratio of 1:10 and stir at 200-300 rpm to form a slurry; heat the slurry to 50-55℃, adjust the pH to 7.0-7.5, add trypsin at 1-2% of the mass of wolfberry extract, and react for 2-4 hours. After enzymatic hydrolysis, the slurry is heated to 85-90℃ and held for 5-10 minutes; then cooled to below 40℃ to obtain wolfberry polypeptide enzymatic hydrolysate. S1.3 Add red algae oligosaccharide iron complex and folic acid to the wolfberry polypeptide hydrolysate, and stir at 300-400 rpm to form an aqueous phase mixture. The astaxanthin-fish oil co-encapsulated preparation was mixed evenly with maltodextrin accounting for 1 / 3 of the total maltodextrin to obtain a powder mixture; S1.4 Add the powder mixture to the aqueous mixture and process it with a high-speed shear disperser at a speed of 10,000-15,000 rpm for 3-5 minutes to form a preliminary emulsion; Then it was transferred to a high-pressure homogenizer and homogenized 2-3 times under a pressure of 30-40MPa to obtain a stable emulsion; The stabilized emulsion was transferred to a vacuum concentration tank and concentrated under conditions of temperature ≤60℃ and vacuum degree ≥-0.08MPa. S1.5. The concentrated slurry is spray-dried, and the dried powder is sieved through an 80-100 mesh sieve to obtain a polypeptide composition for regulating menstruation and nourishing blood.

[0011] Preferably, in step S1.3, the preparation method of the red algae oligosaccharide iron complex is as follows: Pulverize the dried red algae, add deionized water at a material-to-liquid ratio of 1:15-20, extract in a water bath at 80-85℃ for 2-3 hours, filter and collect the supernatant to obtain crude extract of red algae polysaccharides. Adjust the pH of the crude extract of red algae polysaccharides to 4.5-5.5, add cellulase at an enzyme-to-substrate ratio of 1:50-100, and react at 45-50℃ for 6-8 hours to obtain red algae oligosaccharide solution. Ferrous gluconate solution was added to the red algae oligosaccharide solution, and the pH was adjusted to 7.5-8.5 with 0.05 mol / L sodium hydroxide solution. The mixture was stirred at 300-400 rpm for 2-3 hours in a water bath at 60-70℃. After the reaction was completed, the mixture was dialyzed through a dialysis bag with a molecular weight cutoff of 500-1000 Da. The solution was then frozen at -40℃ and freeze-dried under vacuum conditions below 10 Pa to obtain the red algae oligosaccharide iron complex.

[0012] Preferably, the mass ratio of ferrous gluconate to oligosaccharides in the red algae oligosaccharide solution is 1:10-15.

[0013] Preferably, in step S1.5, the solids content of the concentrated slurry is 40%-50%.

[0014] Preferably, in step S1.5, the inlet air temperature is 120-140℃ and the outlet air temperature is 60-70℃.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: In this invention, the polypeptide composition for regulating menstruation and nourishing blood and its preparation method, the astaxanthin-fish oil co-encapsulation preparation uses composite coagulation microencapsulation technology to stably encapsulate the strong antioxidant astaxanthin and fish oil, which can effectively remove excess free radicals generated by physiological stress during menstruation, reduce the inflammatory response of uterine tissue, thereby relieving discomfort symptoms such as dysmenorrhea, and greatly protect unsaturated fatty acids and astaxanthin from oxidative damage, ensuring their biological activity; the red algae oligosaccharide iron complex utilizes oligosaccharides as a natural carrier of iron, and is absorbed through the oligosaccharide iron chelation form in the intestine, bypassing the traditional iron salt absorption pathway, greatly improving bioavailability, and can efficiently and gently supplement the iron necessary for hematopoiesis, improving iron deficiency anemia. Detailed Implementation

[0016] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0017] This invention provides a polypeptide composition for regulating menstruation and nourishing blood, comprising the following raw materials: wolfberry extract, astaxanthin-fish oil co-encapsulation preparation, red algae oligosaccharide iron complex, maltodextrin and folic acid; The astaxanthin-fish oil co-encapsulated formulation is prepared by combining an active oil phase with phospholipids, emulsifying the emulsion through a hydrophobic ceramic membrane, and then encapsulating it using a gelatin-gum arabic composite coagulation method.

[0018] Goji berry extract (50% purity), CAS No.: 11-40-5, purchased from Fufeng Sinote Biotechnology Co., Ltd.; main active ingredients include goji berry polysaccharides, betaine, carotenoids and flavonoids, etc.

[0019] Astaxanthin oil extract (5% purity) was purchased from Hangzhou Linran Biotechnology Co., Ltd.; the main active ingredients include astaxanthin, ketocarotenoids, etc.

[0020] Fish oil (EPA+DHA≥70%) CAS No.: 8016-13-5, purchased from Jishui County Hongxing Natural Medicinal Fragrance Oil Factory.

[0021] Trypsin was purchased from Jinan Yuno Chemical Co., Ltd.; dried red algae was purchased from Qingdao Seaweed Biotechnology Co., Ltd.

[0022] Vitamin E CAS No.: 2074-53-5, Lecithin CAS No.: 8002-43-5, Transglutaminase CAS No.: 80146-85-6, Gelatin CAS No.: 9000-70-8, Gum Arabic CAS No.: 9000-01-5, Ferrous Gluconate CAS No.: 299-29-6, Cellulase CAS No.: 9012-54-8, Maltodextrin CAS No.: 9050-36-6, Folic Acid CAS No.: 59-30-3, purchased from Shanghai Yuanye Biotechnology Co., Ltd.

[0023] Rosemary extract (purity 10%) CAS No.: 84604-14-8, purchased from Jiangsu Aofu Biotechnology Co., Ltd.; the main active ingredients include phenolic diterpenoids, rosmarinic acid, flavonoids (apigenin, hesperidin) and triterpenoids (ursolic acid), etc.

[0024] The composite wall material solution is obtained by mixing a 3% gelatin solution and a 3% gum arabic solution at a mass ratio of 1:1.

[0025] The hydrophobic ceramic membrane is preferably a hydrophobically modified α-alumina tubular microfiltration membrane. The specific process steps are as follows: the cleaned membrane is placed in a vacuum drying oven and dried at 100-120℃ for at least 2 hours; under the protection of dry nitrogen, 1-5% (v / v) of octadecyltrichlorosilane is added to n-hexane and gently stirred to mix evenly; the dried α-alumina membrane is immersed in the octadecyltrichlorosilane solution, the container is sealed, and the membrane is reacted at 40-60℃ for 12-24 hours; after the reaction, the membrane is removed with tweezers and ultrasonically cleaned in fresh anhydrous toluene and anhydrous ethanol for 5-10 minutes each; the cleaned membrane is then placed back in a vacuum drying oven and dried at 60-80℃ for 4-6 hours.

[0026] Example 1: A method for preparing a polypeptide composition for regulating menstruation and nourishing blood, comprising the following steps: S1.1 Weigh the following raw materials in parts by weight: 5 parts by weight of wolfberry extract, 10 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 15 parts by weight of red algae oligosaccharide iron complex, 30 parts by weight of maltodextrin and 0.5 parts by weight of folic acid. S1.2 Add wolfberry extract to deionized water at a volume ratio of 1:10 and stir at 300 rpm to form a slurry; heat the slurry to 50°C, adjust the pH to 7.0, add trypsin at 1% of the mass of wolfberry extract, and react for 4 hours. After enzymatic hydrolysis, the slurry was heated to 85°C and held for 10 minutes; then cooled to below 40°C to obtain wolfberry polypeptide enzymatic hydrolysate. S1.3 Add red algae oligosaccharide iron complex and folic acid to the wolfberry polypeptide hydrolysate, and stir at 400 rpm to form an aqueous mixture. The astaxanthin-fish oil co-encapsulated preparation was mixed evenly with maltodextrin accounting for 1 / 3 of the total maltodextrin to obtain a powder mixture; S1.4 Add the powder mixture to the aqueous mixture and process it with a high-speed shear disperser at 10,000 rpm for 5 minutes to form a preliminary emulsion; It was then transferred to a high-pressure homogenizer and homogenized three times at a pressure of 40 MPa to obtain a stable emulsion. The stabilized emulsion was transferred to a vacuum concentration tank and concentrated to a solids content of 40% under conditions of temperature ≤60℃ and vacuum degree ≥-0.08MPa. S1.5. The concentrated slurry is spray-dried at an inlet air temperature of 120°C and an outlet air temperature of 60°C. The dried powder is then sieved through an 80-mesh sieve to obtain a polypeptide composition for regulating menstruation and nourishing blood.

[0027] The preparation method of red algae oligosaccharide iron complex is as follows: The dried red algae were crushed and added to deionized water at a material-to-liquid ratio of 1:15. The mixture was then extracted in an 80°C water bath for 2 hours. The supernatant was collected by filtration to obtain a crude extract of red algae polysaccharides. The pH of the crude extract of red algae polysaccharides was adjusted to 4.5, and cellulase was added at an enzyme to substrate ratio of 1:50. The mixture was reacted at 45°C for 8 hours to obtain the red algae oligosaccharide solution. Ferrous gluconate solution (mass ratio 1:10) was added to the red algae oligosaccharide solution, and the pH was adjusted to 7.5 with 0.05 mol / L sodium hydroxide solution. The mixture was stirred at 400 rpm for 3 h in a 60 °C water bath. After the reaction was completed, the mixture was dialyzed through a dialysis bag with a molecular weight cutoff of 500 Da. The solution was then frozen at -40 °C and freeze-dried under vacuum conditions below 10 Pa to obtain the red algae oligosaccharide iron complex.

[0028] The preparation method of astaxanthin-fish oil co-encapsulated formulation is as follows: Astaxanthin oil extract and fish oil were mixed at a mass ratio of 1:10. Vitamin E and rosemary extract, which accounted for 0.1% of the total mass of the oil phase, were added. The mixture was stirred at 100 rpm under nitrogen protection at 40°C to obtain the oil phase. The oil phase and lecithin were mixed at a mass ratio of 1:1 and stirred at 200 rpm for 2 hours under nitrogen protection at 50°C to obtain the phospholipid complex oil phase. Tween 80, accounting for 1% of the mass of the phospholipid complex oil phase, was dissolved in deionized water and preheated to 50°C to obtain a continuous phase. The phospholipid complex oil phase was then extruded into the continuous phase through a hydrophobic ceramic membrane with a pore size of 0.8 μm under a pressure of 0.1 MPa to obtain the primary emulsion. The colostrum was added to the composite wall material solution while stirring at 100 rpm, with the mass ratio of the phospholipid complex oil phase to the composite wall material being 1:1. The temperature was maintained at 40℃, and the pH was adjusted to 4.0 by adding 0.1 mol / L citric acid solution dropwise, followed by stirring for 30 min. The temperature was then lowered to 10℃, and 0.05% (by mass) of transglutaminase was added to induce cross-linking for 4 h. After the reaction was completed, the wet microcapsules were collected through a 200-mesh sieve and washed three times with deionized water. The wet microcapsules were then redispersed in deionized water with a solid content of 20%, forming a suspension. The suspension was sprayed into liquid nitrogen to obtain frozen particles; the frozen particles were dried at temperatures below -40°C and below 10 Pa to obtain an astaxanthin-fish oil co-encapsulated formulation.

[0029] Example 2: The difference between this example and Example 1 is that the pH was adjusted to 8.0 using a 0.05 mol / L sodium hydroxide solution.

[0030] Example 3: The difference between this example and Example 1 is that the pH was adjusted to 8.5 using a 0.05 mol / L sodium hydroxide solution.

[0031] Determination of iron content: Accurately weigh a certain mass (m, approximately 0.1 g) of the complex powder into an Erlenmeyer flask, add a mixed acid (e.g., nitric acid-perchloric acid, 10:1, v / v) and heat to digest until the solution is clear and transparent. After cooling, transfer to a volumetric flask (V1, e.g., 100 mL) and dilute to volume. Accurately pipette a certain volume (V2, e.g., 2 mL) of the digestion solution into another volumetric flask (V3, e.g., 50 mL), and add 1 mL of 10% hydroxylamine hydrochloride solution, pH... 5 mL of 4.5% acetate-sodium acetate buffer solution and 2 mL of 0.15% o-phenanthroline solution were diluted to the mark with water, shaken well, and allowed to stand for 15 minutes. The absorbance (A) was measured at 510 nm using a UV-Vis spectrophotometer. The concentration C0 (μg / mL) of the test solution was found from the standard curve (concentration C - absorbance A) plotted with ferrous ammonium sulfate standard solution. The iron content was calculated as follows: Iron content (%) = (C0 × V3 × V1 × 10) / (V3 × V1 × 10) / (V3 × V1 × 10) / (V3 × V1 × 10) / (V3 × V1 × V1 × V2 × V3 × V1 × V2 ... -6 ) / (m×V2)×100%.

[0032] Determination of complexation rate: Accurately weigh a certain mass (m1) of the complex powder, dissolve it in water, and place it in a dialysis bag with a molecular weight cutoff of 500-1000 Da. Dialyze in deionized water for 24 hours, changing the water multiple times during this period to ensure that the free iron is completely dialyzed out. Make up the dialysate to a certain volume (V1), and determine the iron concentration (C) in the dialysate using the method described above for determining iron content. free ); Take another sample of equal mass (m1) accurately, digest it, and determine its total iron content using the same method described above; calculate the complexation rate: Free iron content (%) = (C free ×V1×10 -6 ) / m1×100%; Complexation rate (%)=(1-free iron content% / total iron content%)×100%.

[0033] Table 1 Performance data of red algae oligosaccharide iron complex Iron content Complexation rate Example 1 9.5% 85% Example 2 10.8% 95% Example 3 8.0% 70% Example 2 shows that under weakly alkaline conditions of pH=8.0, both the iron content and complexation rate are relatively high.

[0034] This pH environment ensures that the hydroxyl and carboxyl groups in the red algae oligosaccharides are fully deprotonated, giving them a strong chelating ability; it also maximizes the inhibition of Fe. 2+ Oxidized into Fe 3+ And there is a tendency to form ferric hydroxide precipitate.

[0035] Therefore, iron ions can coordinate with oligosaccharides efficiently and stably to form complete complex structures, resulting in high effective iron content and very few uncomplexed free iron ions.

[0036] In Example 1, although complexation can still occur when the pH is at the lower limit of the range (7.5), the iron content and complexation rate are both lower than those at pH=8.0.

[0037] The alkalinity of the environment was slightly weak, and some coordinating groups were not completely deprotonated, so their complexing ability did not reach the optimal state, resulting in a decrease in reaction efficiency. Some iron ions were not effectively complexed and were therefore removed in the subsequent dialysis purification step, resulting in a decrease in the iron content and complexation rate of the final product.

[0038] In Example 3, when the pH was further increased to 8.5, both the iron content and the complexation rate decreased significantly.

[0039] An excessively high pH environment greatly accelerates the reaction of Fe in the solution. 2+ It reacts with oxygen to produce Fe 3+ The rate of iron precipitate formation is reduced, leading to the formation of a reddish-brown Fe(OH)3 precipitate. These precipitates cannot participate in the complexation reaction, resulting in a decrease in the actual iron source that can form complexes, and thus a significant reduction in the measured iron content.

[0040] Example 4: A method for preparing a polypeptide composition for regulating menstruation and nourishing blood, comprising the following steps: S1.1 Weigh the following raw materials in parts by weight: 15 parts by weight of wolfberry extract, 20 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 25 parts by weight of red algae oligosaccharide iron complex, 50 parts by weight of maltodextrin and 1.5 parts by weight of folic acid. S1.2 Add wolfberry extract to deionized water at a volume ratio of 1:10 and stir at 300 rpm to form a slurry; heat the slurry to 55°C, adjust the pH to 7.5, add trypsin at 2% of the mass of wolfberry extract, and react for 4 hours. After enzymatic hydrolysis, the slurry was heated to 90°C and held for 5 minutes; then cooled to below 40°C to obtain wolfberry polypeptide enzymatic hydrolysate. S1.3 Add red algae oligosaccharide iron complex and folic acid to the wolfberry polypeptide hydrolysate, and stir at 400 rpm to form an aqueous mixture. The astaxanthin-fish oil co-encapsulated preparation was mixed evenly with maltodextrin accounting for 1 / 3 of the total maltodextrin to obtain a powder mixture; S1.4 Add the powder mixture to the aqueous mixture and process it with a high-speed shear disperser at 10,000 rpm for 5 minutes to form a preliminary emulsion; It was then transferred to a high-pressure homogenizer and homogenized three times at a pressure of 40 MPa to obtain a stable emulsion. The stabilized emulsion was transferred to a vacuum concentration tank and concentrated to a solids content of 50% under conditions of temperature ≤60℃ and vacuum degree ≥-0.08MPa. S1.5. The concentrated slurry is spray-dried at an inlet air temperature of 120°C and an outlet air temperature of 60°C. The dried powder is then sieved through a 100-mesh sieve to obtain a polypeptide composition for regulating menstruation and nourishing blood.

[0041] The preparation method of red algae oligosaccharide iron complex is as follows: The dried red algae were crushed and added to deionized water at a material-to-liquid ratio of 1:20. The mixture was then extracted in a water bath at 85°C for 3 hours. The supernatant was collected by filtration to obtain a crude extract of red algae polysaccharides. The pH of the crude extract of red algae polysaccharides was adjusted to 5.5, and cellulase was added at an enzyme to substrate ratio of 1:100. The mixture was reacted at 50°C for 8 hours to obtain the red algae oligosaccharide solution. Ferrous gluconate solution (mass ratio 1:15) was added to the red algae oligosaccharide solution, and the pH was adjusted to 8.0 with 0.05 mol / L sodium hydroxide solution. The reaction was carried out in a 60℃ water bath with stirring at 400 rpm for 2-3 h. After the reaction was completed, the solution was dialyzed through a dialysis bag with a molecular weight cutoff of 1000 Da. The solution was then frozen at -40℃ and freeze-dried under vacuum conditions below 10 Pa to obtain the red algae oligosaccharide iron complex.

[0042] The preparation method of astaxanthin-fish oil co-encapsulated formulation is as follows: Astaxanthin oil extract and fish oil were mixed at a mass ratio of 1:15. Vitamin E and rosemary extract at 0.3% of the total mass of the oil phase and 0.1% of the total mass of the oil phase were added. The mixture was stirred at 100 rpm under nitrogen protection at 40°C to obtain the oil phase. The oil phase and lecithin were mixed at a mass ratio of 1:1.5 and stirred at 200 rpm for 2 hours under nitrogen protection at 60°C to obtain the phospholipid complex oil phase. Tween 80, accounting for 2% of the mass of the phospholipid complex oil phase, was dissolved in deionized water and preheated to 55°C to obtain a continuous phase. The phospholipid complex oil phase was then extruded into the continuous phase through a hydrophobic ceramic membrane with a pore size of 1.2 μm under a pressure of 0.3 MPa to obtain the primary emulsion. The colostrum was added to the composite wall material solution under stirring at 200 rpm, with the mass ratio of the phospholipid complex oil phase to the composite wall material being 1:1.0. The temperature was maintained at 45℃, and 0.1 mol / L citric acid solution was added dropwise to adjust the pH to 4.5, and the mixture was stirred for 45 min. The temperature was then lowered to 10℃, and 0.05% (by mass) of transglutaminase was added to cross-link the colostrum for 4 h. After the reaction was completed, the wet microcapsules were collected through a 200-mesh sieve and washed three times with deionized water. The wet microcapsules were then redispersed in deionized water with a solid content of 30% to form a suspension. The suspension was sprayed into liquid nitrogen to obtain frozen particles; the frozen particles were dried at temperatures below -40°C and below 10 Pa to obtain an astaxanthin-fish oil co-encapsulated formulation.

[0043] Example 5: The difference between this example and Example 4 is that the mass ratio of the phospholipid complex oil phase to the composite wall material is 1:1.5.

[0044] Example 6: The difference between this example and Example 4 is that the mass ratio of the phospholipid complex oil phase to the composite wall material is 1:2.0.

[0045] Example 7: The difference between this example and Example 4 is that the amount of transglutaminase added accounts for 0.07% of the colostrum mass.

[0046] Example 8: The difference between this example and Example 4 is that the amount of transglutaminase added is 0.1% of the colostrum mass.

[0047] Encapsulation efficiency determination: Weigh a certain mass (W) total The sample powder (approximately 0.1 g) was ultrasonically broken up with an appropriate amount of organic solvent (such as n-hexane) to completely dissolve and extract all astaxanthin; the volume was then adjusted to a certain volume (V). totalAfter centrifugation, the supernatant was collected, and its concentration was determined by HPLC (C0). total ), calculate the total astaxanthin mass (M) total =C total ×V total ); and accurately weigh out an equal mass (W) surface Take the powder, add a small amount of mild solvent that cannot penetrate the wall material (such as cold ethanol), gently shake for 1-2 minutes to elute only the free astaxanthin on the surface; after rapid filtration or centrifugation, determine the astaxanthin concentration (C) in the eluent. surface ), calculate the surface astaxanthin mass (M) surface ); Calculate the encapsulation efficiency (%) = (M total -M surface ) / M total ×100%.

[0048] Determination of drug loading: The above steps for determining the total content have yielded the total astaxanthin mass (M). total ) and sample powder mass (W total ); Calculate drug loading (%) = M total / W total ×100%.

[0049] Determination of oxidative stability: The powder was spread evenly in an open petri dish and placed in a constant temperature oven at 60℃; samples were taken every 24 hours; the samples were processed and the POV value was determined according to the method in GB 5009.227-2016 "National Food Safety Standard for Determination of Peroxide Value in Food"; the curve of POV value change over time was plotted to obtain the number of days when the POV value reached 20 meq / kg in the accelerated test at 60℃.

[0050] Table 2 Performance data of astaxanthin-fish oil co-encapsulated formulations Encapsulation efficiency Drug loading Oxidative stability Example 4 85% 18.0% 12 days Example 5 92% 15.5% 18 days Example 6 95% 12.5% 22 days Example 7 89% 18.0% 16 days Example 8 91% 18.0% 25 days Compared with Examples 4, 5, and 6, the encapsulation efficiency was significantly improved as the wall material ratio increased (from 1:1.0 to 1:2.0). This is because more wall material provides more sufficient encapsulation material, which can more completely cover all oil droplets and reduce the exposure of surface oil.

[0051] As the proportion of wall material increases, the drug loading decreases significantly; this is because the proportion of active core (oil phase) in the total product mass decreases, while the proportion of inactive wall material increases.

[0052] Stability increases significantly with the increase of wall material; thicker wall material and higher encapsulation rate together form a more effective physical barrier, better isolating oxygen and slowing down the oxidation rate of the internal oil phase.

[0053] Compared with Examples 4, 7, and 8, the encapsulation efficiency steadily increased as the amount of enzyme increased (from 0.05% to 0.1%). This is because a more complete cross-linking reaction forms a stronger and denser capsule wall, which is more resistant to mechanical damage during subsequent washing and drying processes, reducing the leakage of the core material.

[0054] The drug loading remains essentially unchanged because the cross-linking process is a modification of the wall material and does not change the mass ratio of the core (oil phase) to the wall material in the total feed. Therefore, the content of the effective ingredients in the final product remains unchanged.

[0055] Stability increases dramatically with increasing enzyme content; the covalent cross-linking catalyzed by TG enzyme forms a solid reinforced concrete structure, which is much denser than the brick wall structure that relies solely on electrostatic interaction, and can greatly delay oxygen permeation, thus providing excellent oxidative protection.

[0056] Example 9: A method for preparing a polypeptide composition for regulating menstruation and nourishing blood, comprising the following steps: S1.1 Weigh the following raw materials in parts by weight: 10 parts by weight of wolfberry extract, 10 parts by weight of astaxanthin-fish oil co-encapsulated preparation, 15 parts by weight of red algae oligosaccharide iron complex, 40 parts by weight of maltodextrin and 1.0 part by weight of folic acid. S1.2 Add wolfberry extract to deionized water at a volume ratio of 1:10 and stir at 300 rpm to form a slurry; heat the slurry to 50°C, adjust the pH to 7.0, add trypsin at 1% of the mass of wolfberry extract, and react for 4 hours. After enzymatic hydrolysis, the slurry was heated to 90°C and held for 5 minutes; then cooled to below 40°C to obtain wolfberry polypeptide enzymatic hydrolysate. S1.3 Add red algae oligosaccharide iron complex and folic acid to the wolfberry polypeptide hydrolysate, and stir at 400 rpm to form an aqueous mixture. The astaxanthin-fish oil co-encapsulated preparation was mixed evenly with maltodextrin accounting for 1 / 3 of the total maltodextrin to obtain a powder mixture; S1.4 Add the powder mixture to the aqueous mixture and process it with a high-speed shear disperser at 10,000 rpm for 5 minutes to form a preliminary emulsion; It was then transferred to a high-pressure homogenizer and homogenized three times at a pressure of 40 MPa to obtain a stable emulsion. The stabilized emulsion was transferred to a vacuum concentration tank and concentrated to a solids content of 45% under conditions of temperature ≤60℃ and vacuum degree ≥-0.08MPa. S1.5. The concentrated slurry is spray-dried at an inlet air temperature of 120°C and an outlet air temperature of 60°C. The dried powder is then sieved through an 80-mesh sieve to obtain a polypeptide composition for regulating menstruation and nourishing blood.

[0057] The preparation method of red algae oligosaccharide iron complex is as follows: The dried red algae were crushed and added to deionized water at a ratio of 1:15. The mixture was then extracted in a water bath at 80°C for 3 hours. The supernatant was collected by filtration to obtain a crude extract of red algae polysaccharides. The pH of the crude extract of red algae polysaccharides was adjusted to 5.0, and cellulase was added at an enzyme to substrate ratio of 1:80. The mixture was reacted at 45°C for 8 hours to obtain the red algae oligosaccharide solution. Ferrous gluconate solution (mass ratio 1:12) was added to the red algae oligosaccharide solution, and the pH was adjusted to 8.0 with 0.05 mol / L sodium hydroxide solution. The reaction was carried out in a 60℃ water bath with stirring at 300 rpm for 3 h. After the reaction was completed, the solution was dialyzed through a dialysis bag with a molecular weight cutoff of 500 Da. The solution was then frozen at -40℃ and freeze-dried under vacuum conditions below 10 Pa to obtain the red algae oligosaccharide iron complex.

[0058] The preparation method of astaxanthin-fish oil co-encapsulated formulation is as follows: Astaxanthin oil extract and fish oil were mixed at a mass ratio of 1:12. Vitamin E and rosemary extract, which accounted for 0.2% of the total mass of the oil phase, were added. The mixture was stirred at 100 rpm under nitrogen protection at 40°C to obtain the oil phase. The oil phase and lecithin were mixed at a mass ratio of 1:1.2 and stirred at 200 rpm for 2 hours under nitrogen protection at 60°C to obtain the phospholipid complex oil phase. Tween 80, accounting for 2% of the mass of the phospholipid complex oil phase, was dissolved in deionized water and preheated to 55°C to obtain a continuous phase. The phospholipid complex oil phase was then extruded into the continuous phase through a hydrophobic ceramic membrane with a pore size of 1.0 μm under a pressure of 0.3 MPa to obtain the primary emulsion. The colostrum was added to the composite wall material solution while stirring at 200 rpm, with the mass ratio of the phospholipid complex oil phase to the composite wall material being 1:1.5. The temperature was maintained at 45℃, and 0.1 mol / L citric acid solution was added dropwise to adjust the pH to 4.5, and the mixture was stirred for 45 min. The temperature was then lowered to 10℃, and 0.1% of transglutaminase by the mass of the colostrum was added for cross-linking for 4 h. After the reaction was completed, the wet microcapsules were collected through a 200-mesh sieve and washed three times with deionized water. The wet microcapsules were then redispersed in deionized water with a solid content of 30% to form a suspension. The suspension was sprayed into liquid nitrogen to obtain frozen particles; the frozen particles were dried at temperatures below -40°C and below 10 Pa to obtain an astaxanthin-fish oil co-encapsulated formulation.

[0059] Example 10: The difference between this example and Example 9 is that 15 parts by weight of the astaxanthin-fish oil co-encapsulated preparation are used.

[0060] Example 11: The difference between this example and Example 9 is that 20 parts by weight of the astaxanthin-fish oil co-encapsulated preparation are used.

[0061] Example 12: The difference between this example and Example 9 is that 20 parts by weight of red algae oligosaccharide iron complex are used.

[0062] Example 13: The difference between this example and Example 9 is that 25 parts by weight of red algae oligosaccharide iron complex are used.

[0063] Determination of active ingredient content: Accurately weigh a certain mass (W) ast The complex powder (approximately 0.1 g) was subjected to vortexing and ultrasonic extraction with an appropriate amount of chloroform-methanol (2:1, v / v) mixed solvent to fully disrupt the structure and extract astaxanthin; after centrifugation, the supernatant was taken and diluted to a certain volume (V). ast Using a UV-Vis spectrophotometer with n-hexane as the solvent, the absorbance was measured at a wavelength of 480 nm. The content of astaxanthin was calculated based on its standard curve and extinction coefficient: Astaxanthin content (%) = (C... ast ×V ast ) / W ast ×100%.

[0064] Determination of solubility and dispersibility: Measure 100 mL of warm water (40℃±2℃) into a graduated beaker, weigh 5.0 g of product powder, add it all at once from the center of the vortex, and stir at 300 rpm; stop stirring, let stand for 30 minutes, and observe and record whether there is precipitation, layering or floating oil.

[0065] Table 3 Performance data of the peptide composition for regulating menstruation and nourishing blood Astaxanthin content Iron content Oxidative stability Precipitation rate Example 9 0.50% 2.25% 18 days 5.0% Example 10 0.75% 2.25% 15 days 6.5% Example 11 1.00% 2.25% 12 days 8.0% Example 12 0.50% 3.00% 17 days 4.0% Example 13 0.50% 3.75% 16 days 3.5% With the increase of the amount of astaxanthin-fish oil co-encapsulated formulation added (Examples 9, 10, 11), the final astaxanthin content in the product increased linearly; similarly, the increase of the amount of red algae oligosaccharide iron complex added (Examples 9, 12, 13) directly led to a linear increase in the iron content of the product.

[0066] Increasing the oil phase content reduces stability (Examples 9, 10, 11). When the weight of the astaxanthin-fish oil co-encapsulated formulation increases, the oxidative stability decreases significantly. This is because although the co-encapsulated formulation itself has a protective layer, the total amount of highly unsaturated fatty acids in the product increases, which means more easily oxidized substrates. At the same time, the high proportion of oily components will slightly affect the integrity of the wall material, making the overall product more susceptible to oxidation.

[0067] Increasing the iron content slightly affects stability (Examples 9, 12, 13). Although the iron in the red algae oligosaccharide iron complex has been complexed, greatly improving stability, increasing the amount added still introduces a slight risk of prooxidation, thus resulting in a slight decrease in stability.

[0068] Increasing the oil phase content will worsen the dispersibility (Examples 9, 10, 11). Astaxanthin-fish oil co-encapsulated formulations are hydrophobic oily powders. The higher the amount added, the stronger the overall hydrophobicity of the product. When added to water, these hydrophobic particles tend to aggregate and precipitate, resulting in an increased precipitation rate and poor dispersibility.

[0069] Increasing the content of iron complexes improves dispersibility (Examples 9, 12, 13). Red algae oligosaccharide iron complexes are hydrophilic components. Increasing their addition is equivalent to increasing the proportion of hydrophilic components in the product, thus making it easier to disperse evenly in water, reducing the sedimentation rate, and improving dispersibility.

[0070] Considering the astaxanthin content, iron content, stability, and solubility, Example 10, while ensuring the content of significant functional components, also possesses acceptable stability and dispersibility. Therefore, Example 10 is selected as the optimal example. Comparative Example 1: The difference between this example and Example 10 is that no astaxanthin-fish oil co-encapsulated formulation was added.

[0071] Comparative Example 2: The difference between this example and Example 10 is that no red algae oligosaccharide iron complex was added.

[0072] In vitro antioxidant activity assay: Trolox standard working solution and the test sample solution were diluted according to a predetermined gradient. 150 μL of sodium fluorescein working solution (70 nM) and 25 μL of Trolox standard solution or the test sample solution were added sequentially to each well of a 96-well black fluorescent plate (25 μL of PBS buffer was added to the blank control wells). The plate was gently shaken to mix. The plate was placed in a preheated fluorescence detector and incubated at 37°C for 10 minutes to allow temperature equilibration. After incubation, 25 μL of the solution was quickly added to each well using an autosampler or manually. Immediately begin monitoring fluorescence changes using AAPH solution (12 mM, final concentration approximately 12 mM); for each well (including blank and sample), calculate the total area under the fluorescence decay curve (AUC) using instrument software or manually; plot a standard curve with the concentrations of different Trolox standards on the x-axis and their corresponding net AUC values ​​on the y-axis; calculation formula: ORAC value (μmolTE / g) = (C × V × D) / M, where: C is the Trolox equivalent concentration (μM) calculated from the standard curve, V is the total volume of the sample solution (L), D is the dilution factor of the sample solution, and M is the mass of the sample (g).

[0073] Table 4 Performance data of the peptide composition for regulating menstruation and nourishing blood Astaxanthin content Iron content Oxidative stability Precipitation rate ORAC value Example 10 0.75% 2.25% 15 days 6.5% 850.5 μmol TE / g Comparative Example 1 0% 2.25% > 28 days 2.0% 105.2 μmol TE / g Comparative Example 2 0.75% 0% 20 days 9.5% 780.3 μmol TE / g Analysis of Comparative Example 1 (prepared formulation without added astaxanthin-fish oil): The astaxanthin content was 0%, completely losing the anti-inflammatory and antioxidant effects of this ingredient, and the antioxidant capacity decreased sharply, while the iron content remained unchanged.

[0074] It has excellent stability (>28 days), but under this condition it has lost its main core functions such as anti-oxidation and cannot meet the purpose of regulating menstruation and nourishing blood.

[0075] It exhibits the best dispersibility (sedimentation rate of only 2.0%) because the hydrophobic oily powder has been removed, resulting in a product with extremely strong overall hydrophilicity that can quickly and completely disperse and dissolve in water.

[0076] Comparative Example 2 (without added red algae oligosaccharide iron complex): The iron content was 0%, completely losing its key blood-nourishing effects, while the astaxanthin content remained unchanged.

[0077] The stability was better than that of the whole formulation, which indicates that the iron element in the red algae oligosaccharide iron complex, although it has been complexed by oligosaccharides, still has a slight negative impact on the overall stability of the system due to its inherent pro-oxidative properties. Removing it improves the stability.

[0078] The product exhibited the worst dispersibility (9.5% sedimentation rate). This is because the red algae oligosaccharide iron complex itself has good water solubility. Its absence leads to a relatively higher proportion of hydrophobic components (astaxanthin-fish oil co-encapsulated formulation) in the product, disrupting the hydrophilic-hydrophobic balance and causing more oily powder to aggregate and precipitate.

[0079] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely preferred examples and are not intended to limit the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A polypeptide composition for regulating menstruation and nourishing blood, characterized in that, It includes the following ingredients: wolfberry extract, astaxanthin-fish oil co-encapsulation preparation, red algae oligosaccharide iron complex, maltodextrin and folic acid; The astaxanthin-fish oil co-encapsulated formulation is prepared by combining an active oil phase with phospholipids, emulsifying the emulsion through a hydrophobic ceramic membrane, and then encapsulating it using a gelatin-gum arabic composite coagulation method.

2. The polypeptide composition for regulating menstruation and nourishing blood according to claim 1, characterized in that, The mixture consists of 5-15 parts by weight of wolfberry extract, 10-20 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 15-25 parts by weight of red algae oligosaccharide iron complex, 30-50 parts by weight of maltodextrin, and 0.5-1.5 parts by weight of folic acid.

3. The polypeptide composition for regulating menstruation and nourishing blood according to claim 2, characterized in that, The preparation method of the astaxanthin-fish oil co-encapsulated formulation is as follows: Astaxanthin oil extract and fish oil were mixed at a mass ratio of 1:10-15, and vitamin E and rosemary extract were added. The mixture was stirred at 50-100 rpm under nitrogen protection at 30-40°C to obtain the oil phase. The oil phase and lecithin were mixed at a mass ratio of 1:1-1.5 and stirred at 100-200 rpm for 1-2 hours under nitrogen protection at 50-60℃ to obtain the phospholipid complex oil phase. Dissolve 1-2% of Tween 80 (by mass of the phospholipid complex oil phase) in deionized water and preheat to 50-55°C to obtain a continuous phase. Extrude the phospholipid complex oil phase into the continuous phase through a hydrophobic ceramic membrane with a pore size of 0.8-1.2 μm under a pressure of 0.1-0.3 MPa to obtain the primary emulsion. The colostrum was added to the composite wall material solution while stirring at 100-200 rpm, with the mass ratio of the phospholipid complex oil phase to the composite wall material being 1:1.0-2.

0. The temperature was maintained at 40-45℃, and 0.1 mol / L citric acid solution was added dropwise to adjust the pH to 4.0-4.5, followed by stirring for 30-45 minutes. The temperature was lowered to 5-10℃, and 0.05-0.1% (by mass) of transglutaminase was added for cross-linking for 2-4 hours. After the reaction was complete, the wet microcapsules were collected using a 200-mesh sieve and washed 2-3 times with deionized water. The wet microcapsules were redispersed in deionized water with a solid content of 20%-30%, forming a suspension. The suspension was sprayed into liquid nitrogen to obtain frozen particles; the frozen particles were dried at temperatures below -40°C and below 10 Pa to obtain an astaxanthin-fish oil co-encapsulated formulation.

4. The polypeptide composition for regulating menstruation and nourishing blood according to claim 3, characterized in that, The amount of vitamin E added is 0.1-0.3% of the total mass of the oil phase, and the amount of rosemary extract added is 0.05-0.1% of the total mass of the oil phase.

5. The polypeptide composition for regulating menstruation and nourishing blood according to claim 3, characterized in that, The composite wall material solution is obtained by mixing a 3% gelatin solution and a 3% gum arabic solution at a mass ratio of 1:

1.

6. A method for preparing a menstrual regulating and blood-nourishing polypeptide composition, used to prepare the menstrual regulating and blood-nourishing polypeptide composition as described in any one of claims 1-5, characterized in that, The preparation method of the menstrual regulating and blood-nourishing polypeptide composition is as follows: S1.1 Weigh the following raw materials in parts by weight: 5-15 parts by weight of wolfberry extract, 10-20 parts by weight of astaxanthin-fish oil co-encapsulation preparation, 15-25 parts by weight of red algae oligosaccharide iron complex, 30-50 parts by weight of maltodextrin and 0.5-1.5 parts by weight of folic acid. S1.2 Add wolfberry extract to deionized water at a volume ratio of 1:10 and stir at 200-300 rpm to form a slurry; heat the slurry to 50-55℃, adjust the pH to 7.0-7.5, add trypsin at 1-2% of the mass of wolfberry extract, and react for 2-4 hours. After enzymatic hydrolysis, the slurry is heated to 85-90℃ and held for 5-10 minutes; then cooled to below 40℃ to obtain wolfberry polypeptide enzymatic hydrolysate. S1.3 Add red algae oligosaccharide iron complex and folic acid to the wolfberry polypeptide hydrolysate, and stir at 300-400 rpm to form an aqueous phase mixture. The astaxanthin-fish oil co-encapsulated preparation was mixed evenly with maltodextrin accounting for 1 / 3 of the total maltodextrin to obtain a powder mixture; S1.4 Add the powder mixture to the aqueous mixture and process it with a high-speed shear disperser at a speed of 10,000-15,000 rpm for 3-5 minutes to form a preliminary emulsion; Then it was transferred to a high-pressure homogenizer and homogenized 2-3 times under a pressure of 30-40MPa to obtain a stable emulsion; The stabilized emulsion was transferred to a vacuum concentration tank and concentrated under conditions of temperature ≤60℃ and vacuum degree ≥-0.08MPa. S1.

5. The concentrated slurry is spray-dried, and the dried powder is sieved through an 80-100 mesh sieve to obtain a polypeptide composition for regulating menstruation and nourishing blood.

7. The method for preparing the menstrual regulating and blood-nourishing polypeptide composition according to claim 6, characterized in that, In step S1.3, the preparation method of the red algae oligosaccharide iron complex is as follows: Pulverize the dried red algae, add deionized water at a material-to-liquid ratio of 1:15-20, extract in a water bath at 80-85℃ for 2-3 hours, filter and collect the supernatant to obtain crude extract of red algae polysaccharides. Adjust the pH of the crude extract of red algae polysaccharides to 4.5-5.5, add cellulase at an enzyme-to-substrate ratio of 1:50-100, and react at 45-50℃ for 6-8 hours to obtain red algae oligosaccharide solution. Ferrous gluconate solution was added to the red algae oligosaccharide solution, and the pH was adjusted to 7.5-8.5 with 0.05 mol / L sodium hydroxide solution. The mixture was stirred at 300-400 rpm for 2-3 hours in a water bath at 60-70℃. After the reaction was completed, the mixture was dialyzed through a dialysis bag with a molecular weight cutoff of 500-1000 Da. The solution was then frozen at -40℃ and freeze-dried under vacuum conditions below 10 Pa to obtain the red algae oligosaccharide iron complex.

8. The method for preparing the menstrual regulating and blood-nourishing polypeptide composition according to claim 7, characterized in that, The mass ratio of ferrous gluconate to oligosaccharides in the red algae oligosaccharide solution is 1:10-15.

9. The method for preparing the menstrual regulating and blood-nourishing polypeptide composition according to claim 6, characterized in that, In S1.5, the solids content of the concentrated slurry is 40%-50%.

10. The method for preparing the menstrual regulating and blood-nourishing polypeptide composition according to claim 6, characterized in that, In S1.5, the inlet air temperature is 120-140℃ and the outlet air temperature is 60-70℃.