Anti-aging composition containing paeonia ostii peptide and preparation method of anti-aging composition

Abstract

The invention discloses an anti-aging composition containing paeonia ostii flower peptide and a preparation method thereof.The anti-aging composition is prepared from, by mass, 9-11 parts of paeonia ostii flower peptide, 4-6 parts of compound plant flavone, 1.5-2 parts of chitosan oligosaccharide, 0.4-0.5 part of sodium ascorbyl phosphate and 0.5-1 part of microencapsulated beta-glucan. The method comprises the following steps: extracting protein through an isoelectric precipitation method, carrying out continuous flow ultrasonic-enzymolysis, and finally carrying out three-stage membrane treatment on a paeonia ostii peptide stock solution prepared in S12 to obtain the paeonia ostii peptide, so that a strong alkaline environment is avoided, the protein yield is increased by 12%, the enzymolysis efficiency is increased by 40%, the time is shortened to 2 hours, the energy consumption is reduced by 67%, and the activity retention rate is greater than 98%; through subcritical water extraction, the total yield of flavonoids is increased from 65% to 89%, ethanol and a surfactant are completely eliminated, the extraction period is shortened from 2 hours to 15 minutes, then polysaccharide and pigment impurities are removed through stepped elution, the composite plant flavonoids are prepared, and the purity of the total flavonoids is increased from 65% to 92%.

Description

Technical Field

[0001] The invention relates to the technical field of compositions, and in particular to an anti-aging composition containing peony flower peptide and a preparation method thereof. Background Art

[0002] Aging is a continuous process of change. The international mainstream believes that free radicals damage human cells, thus causing many diseases and accelerating human aging. From generation to quenching, free radicals have a profound impact on the formation of lipofuscin, mutation of mitochondrial DNA, induction of cell apoptosis and protein synthesis, the relationship between free radicals and aging, and the diseases that free radicals can cause. Therefore, how to delay aging has been proposed, providing a certain scientific basis for humans to slow down their own aging. Anti-aging does not mean to increase people's lifespan, but to fight against the factors that shorten lifespan caused by the continuous generation of free radicals in the human body due to continuous contact with the outside world, including breathing (oxidation reaction), external pollution, radiation exposure and other factors.

[0003] According to the publication number: CN118286384A-An anti-aging composition containing Danfeng peony flower peptide and its preparation method, the reference patent has the following problems: 1. Risk of damage during protein extraction. The strong alkaline environment of pH 8-10 is used to extract protein, which may cause oxidation and degradation of some sensitive amino acids (such as tryptophan), affecting the biological activity of the final product. The enzyme step uses 80-90°C, which may denature some heat-sensitive peptides (such as peptide chains containing thiol groups) and reduce antioxidant activity; 2. Ingredient stability and compatibility issues. Chitosan oligosaccharides (cationic polysaccharides) are not suitable for the extraction of proteins. Sugar) and sodium ascorbyl phosphate (anion) may combine through electrostatic interaction to form a precipitate. Trehalose only protects the peptide segment and does not take into account the oxidative sensitivity of the composite flavonoids. The flavonoids extracted with 40% ethanol are easily degraded by moisture absorption after drying; 3. Bottleneck of large-scale production: ultrasonic pretreatment has high energy consumption and lasts for 20-30 minutes at a power of 600-700W. The heat dissipation and energy consumption costs of the equipment increase dramatically during large-scale production. Moreover, both Danfeng peony peptide and composite flavonoids adopt freeze-drying process, and the energy consumption per kilogram of material is 3-5 times higher than that of spray drying, which has a high cost and affects large-scale production. Summary of the invention

[0004] In order to overcome the above-mentioned shortcomings, the present invention provides an anti-aging composition containing Danfeng peony flower peptide and a preparation method thereof.

[0005] The present invention achieves the above-mentioned purpose through the following technical solutions:

[0006] An anti-aging composition containing phoenix peony flower peptide comprises the following raw materials by weight: 9-11 parts of phoenix peony flower peptide, 4-6 parts of compound plant flavonoids, 1.5-2 parts of chitosan oligosaccharide, 0.4-0.5 parts of sodium ascorbyl phosphate and 0.5-1 parts of microencapsulated beta-glucan.

[0007] Preferably, the preparation method of the Danfeng peony flower peptide is as follows:

[0008] S11. Using Danfeng peony flowers as raw materials, extract proteins by the isoelectric point precipitation method. Mix Danfeng peony flower proteins with water at a ratio of 1:25 (g / mL) to obtain a Danfeng peony flower protein solution. Avoid a strongly alkaline environment, and the protein yield is increased by 12%;

[0009] S12. Continuous flow ultrasonic-enzymatic hydrolysis. Subject the Danfeng peony flower protein solution obtained in step S11 to ultrasonic pretreatment, composite enzyme hydrolysis, enzyme inactivation, cooling, and centrifugation to obtain a crude Danfeng peony flower peptide solution;

[0010] S13. Subject the crude Danfeng peony flower peptide solution obtained in S12 to three-stage membrane treatment to obtain Danfeng peony flower peptide.

[0011] Preferably, the ultrasonic pretreatment uses a 50 kHz high-frequency continuous flow device with a power density of 15 W / mL and a treatment time of 10 minutes.

[0012] Preferably, in the composite enzyme hydrolysis and enzyme inactivation, add acid protease and flavor protease to the Danfeng peony flower protein solution that has undergone ultrasonic pretreatment, adjust to pH = 5.0 (the intersection point of the optimal pH of the two enzymes), and then carry out a dynamic circulation reaction at 55 °C. The degree of hydrolysis reaches 35%, the enzymatic hydrolysis efficiency is increased by 40%, and the time is shortened to 2 h.

[0013] Preferably, in step S13, first remove the unhydrolyzed proteins through a 100 kDa ultrafiltration membrane, then enrich the active peptide segments through a 10 kDa nanofiltration membrane, and finally concentrate to a solid content of ≥25% through a reverse osmosis membrane. The energy consumption is reduced by 67%, and the activity retention rate is >98%.

[0014] Preferably, the preparation method of the composite plant flavonoids is as follows:

[0015] S21. Subcritical water extraction. Crush Galinsoga parviflora, Taraxacum mongolicum, and Perilla frutescens leaves to obtain a composite plant powder; then add subcritical water at a ratio of 40:1 of subcritical water to the composite plant powder, maintain at a temperature of 120 °C ± 5 (higher than the boiling point of water but lower than the flavonoid decomposition temperature) and a pressure of 3 MPa (maintaining the liquid water state) for 15 minutes, and then add 0.1% β-cyclodextrin to encapsulate the thermosensitive components to obtain flavonoids. The total flavonoid yield is increased from 65% to 89%, ethanol and surfactants are completely eliminated, and the extraction cycle is compressed from 2 hours to 15 minutes;

[0016] S22. Macroporous resin purification. Select AB-8 type resin (specific surface area 580 m 2Dynamic adsorption was carried out (average pore size 13.5 nm), the sample loading flow rate was 2 BV / h (1 BV = resin volume), the sample loading concentration was 1.2 mg flavonoids / mL, the adsorption rate was > 95%, and then through stepwise elution, polysaccharides and pigment impurities were removed, and finally compound plant flavonoids were obtained.

[0017] Preferably, the mixing ratio of the ground Galinsoga parviflora, Taraxacum mongolicum and Perilla frutescens leaves is 5:3:2, and it is ultrafinely ground to D90 ≤ 50 μm to increase the specific surface area.

[0018] Preferably, the stepwise elution includes pretreatment elution, main elution and fine elution. The pretreatment elution is carried out at a flow rate of 3 BV / h in an ethanol solution with a concentration of 30% to remove polysaccharides. The main elution is carried out at a flow rate of 1.5 BV / h in an ethanol solution with a concentration of 70% to remove quercetin / luteolin. The fine elution is carried out at a flow rate of 2 BV / h in an ethanol solution with a concentration of 90% to remove apigenin.

[0019] A preparation method of an anti-aging composition containing Paeonia ostii flower peptide as described above. Weigh Paeonia ostii flower peptide, compound plant flavonoids, chitosan oligosaccharide, sodium ascorbyl phosphate and microencapsulated β-glucan according to mass parts, and mix them evenly to obtain.

[0020] Use of the anti-aging composition containing Paeonia ostii flower peptide as described above in the preparation of anti-aging products.

[0021] The beneficial effects of the present invention are as follows: In the anti-aging composition containing Paeonia ostii flower peptide and its preparation method,

[0022] 1. Using Paeonia ostii flower as the raw material, proteins are extracted by the isoelectric point precipitation method, followed by continuous flow ultrasonic-enzymatic hydrolysis. Finally, the Paeonia ostii flower peptide stock solution prepared from S12 is subjected to three-stage membrane treatment to obtain Paeonia ostii flower peptide. This not only avoids a strong alkaline environment, increases the protein yield by 12%, but also improves the enzymatic hydrolysis efficiency by 40%, shortens the time to 2 h, reduces the energy consumption by 67% at the same time, and the activity retention rate is > 98%;

[0023] 2. Through subcritical water extraction, the total flavonoid yield is increased from 65% to 89%, ethanol and surfactants are completely eliminated, the extraction cycle is compressed from 2 hours to 15 minutes, and then through stepwise elution, polysaccharides and pigment impurities are removed, and finally compound plant flavonoids are obtained, and the total flavonoid purity is increased from 65% to 92%. Detailed Description of the Invention

[0024] Now, the present invention will be further described in detail.

[0025] Example 1:

[0026] An anti-aging composition containing peony flower peptides from Danfeng, calculated by mass fraction, the raw materials include the following components: 9-11 parts of peony flower peptides from Danfeng, 4-6 parts of compound plant flavonoids, 1.5-2 parts of chitosan oligosaccharide, 0.4-0.5 parts of sodium ascorbyl phosphate, and 0.5-1 of microencapsulated β-glucan.

[0027] Compared with the components of the reference patent, adjustments have been made respectively.

[0028] Ingredients Reference Patent This Patent Basis for Improvement Danfeng Peony Flower Peptide 8-10 9-11 Increase the proportion of active ingredients to enhance efficacy Compound Plant Flavonoids 3-5 4-6 Add the synergistic effect of flavonoids Chitosan oligosaccharide 1-2 1.5-2 Limit the molecular weight ≤ 3000Da to enhance transdermal absorption Sodium Ascorbyl Phosphate 0.3-0.5 0.4-0.5 Improve stability Microencapsulated β-glucan 0.5-1 New ingredient to enhance ingredient stability

[0029] Specifically, the preparation method of the peony flower peptides from Danfeng is as follows:

[0030] S11. Using peony flowers from Danfeng as raw materials, extract proteins by the isoelectric point precipitation method. Mix peony flower proteins from Danfeng with water at a ratio of 1:25 (g / mL) to obtain a peony flower protein solution from Danfeng, avoiding a strong alkaline environment, and the protein yield is increased by 12%;

[0031] S12. Continuous flow ultrasonic-enzymatic hydrolysis. The peony flower protein solution in step S11 is subjected to ultrasonic pretreatment, composite enzyme hydrolysis, enzyme inactivation, cooling, and centrifugation to obtain a crude solution of peony flower peptides from Danfeng;

[0032] S13. Subject the crude solution of peony flower peptides from Danfeng prepared in S12 to three-stage membrane treatment to obtain peony flower peptides from Danfeng.

[0033] Specifically, the ultrasonic pretreatment uses a 50 kHz high-frequency continuous flow device with a power density of 15 W / mL and a treatment time of 10 minutes.

[0034] Specifically, in the composite enzyme hydrolysis and enzyme inactivation, add acid protease and flavor protease to the peony flower protein solution after ultrasonic pretreatment, adjust to pH = 5.0 (the intersection point of the optimal pH of the two enzymes), and then carry out a dynamic circulation reaction at 55 °C. The degree of hydrolysis reaches 35%, the enzymatic hydrolysis efficiency is increased by 40%, and the time is shortened to 2 h.

[0035] Specifically, in step S13, first remove the unhydrolyzed proteins through a 100 kDa ultrafiltration membrane, then enrich the active peptide segments through a 10 kDa nanofiltration membrane, and finally concentrate to a solid content of ≥25% through a reverse osmosis membrane. The energy consumption is reduced by 67%, and the activity retention rate is >98%.

[0036] Specifically, the preparation method of the compound plant flavonoids is as follows:

[0037] S21. Subcritical water extraction: Grind galinsoga parviflora, dandelion, and perilla leaves to obtain a composite plant powder. Then, add subcritical water at a ratio of 40:1 to the composite plant powder, maintain it at a temperature of 120°C ± 5 (higher than the boiling point of water but lower than the flavonoid decomposition temperature) and a pressure of 3 MPa (to maintain the liquid water state) for 15 minutes. Then, add 0.1% β-cyclodextrin to encapsulate heat-sensitive components to obtain flavonoids. The total yield of flavonoids is increased from 65% to 89%, ethanol and surfactants are completely eliminated, and the extraction cycle is compressed from 2 hours to 15 minutes.

[0038] S22. Macroporous resin purification: Select AB-8 type resin (specific surface area 580 m 2 / g, average pore diameter 13.5 nm) for dynamic adsorption. The sample loading flow rate is 2 BV / h (1 BV = resin volume), the sample loading concentration is 1.2 mg flavonoids / mL, and the adsorption rate is > 95%. Then, perform stepwise elution to remove polysaccharides and pigment impurities, and finally obtain composite plant flavonoids.

[0039] Specifically, the mixing ratio of the ground galinsoga parviflora, dandelion, and perilla leaves is 5:3:2, and it is ultra-finely ground to D90 ≤ 50 μm to increase the specific surface area.

[0040] Specifically, the stepwise elution includes pretreatment elution, main elution, and fine elution. The pretreatment elution is performed in a 30% ethanol solution at a flow rate of 3 BV / h to remove polysaccharides. The main elution is performed in a 70% ethanol solution at a flow rate of 1.5 BV / h to remove quercetin / luteolin. The fine elution is performed in a 90% ethanol solution at a flow rate of 2 BV / h to remove apigenin.

[0041] A preparation method of an anti-aging composition containing peony flower peptides as described above. Weigh peony flower peptides, composite plant flavonoids, chitosan oligosaccharide, sodium ascorbyl phosphate, and microencapsulated β-glucan by mass fraction and mix them evenly to obtain the composition.

[0042] Comparative Example 1:

[0043] An anti-aging composition containing peony flower peptides. By mass fraction, the raw materials include the following components: 9 parts of peony flower peptides, 4 parts of composite plant flavonoids, 1.5 parts of chitosan oligosaccharide, 0.4 part of sodium ascorbyl phosphate, and 0.5 part of microencapsulated β-glucan. The preparation method is the same as that in Example 1.

[0044] Comparative Example 2:

[0045] An anti-aging composition containing Danfeng peony flower peptide, by mass, the raw materials include the following components: 10 parts of Danfeng peony flower peptide, 5 parts of complex plant flavonoids, 1.5 parts of chitosan oligosaccharide, 0.4 part of sodium ascorbyl phosphate, and 0.6 part of microencapsulated β-glucan. The preparation method is as the preparation method in Example 1.

[0046] Comparative Example 3:

[0047] An anti-aging composition containing Danfeng peony flower peptide, by mass, the raw materials include the following components: 11 parts of Danfeng peony flower peptide, 6 parts of complex plant flavonoids, 2 parts of chitosan oligosaccharide, 0.5 part of sodium ascorbyl phosphate, and 1 part of microencapsulated β-glucan. The preparation method is as the preparation method in Example 1.

[0048] The experimental comparison is as follows:

[0049] DPPH free radical scavenging rate comparison (2mg / mL solution)

[0050] Sample Clearance Rate (%) Comparative Example 1 76.5±2.1 Comparative Example 2 81.7±1.8 Comparative Example 3 84.6±1.5

[0051] Stability test

[0052]

[0053] In summary, the anti-aging composition containing Danfeng peony flower peptide of the present invention can play an anti-aging role.

[0054] Based on the inspiration of the present invention, through the above description, relevant staff can completely make various changes and modifications without departing from the technical idea of the present invention. The technical scope of the present invention is not limited to the content in the specification, and its technical scope must be determined according to the scope of the claims.

Claims

1. An anti-aging composition containing Paeonia ostii flower peptide, characterized in that: By mass parts, the raw materials include the following components: 9 - 11 parts of Danfeng peony flower peptide, 4 - 6 parts of compound plant flavonoids, 1.5 - 2 parts of chitosan oligosaccharide, 0.4 - 0.5 parts of sodium ascorbyl phosphate, and 0.5 - 1 part of microencapsulated β - glucan.

2. The anti-aging composition containing Paeonia ostii T. Hong & J. X. Zhang flower peptide according to claim 1, wherein: The preparation method of the Danfeng peony flower peptide is as follows: S11. Using Danfeng peony flower as the raw material, extract the protein by the isoelectric point precipitation method. Mix the Danfeng peony flower protein with water at a ratio of 1:25 (g / mL) to obtain the Danfeng peony flower protein solution. S12. Continuous - flow ultrasonic - enzymatic hydrolysis. Subject the Danfeng peony flower protein solution in step S11 to ultrasonic pretreatment, composite enzyme hydrolysis, enzyme inactivation, cooling, and centrifugation to obtain the crude Danfeng peony flower peptide solution. S13. Subject the crude Danfeng peony flower peptide solution prepared in S12 to three - stage membrane treatment to obtain the Danfeng peony flower peptide.

3. The anti-aging composition containing Paeonia ostii Feng Dan flower peptide according to claim 2, wherein: The ultrasonic pretreatment uses a 50 kHz high - frequency continuous - flow device with a power density of 15 W / mL and a treatment time of 10 minutes.

4. The anti-aging composition containing Paeonia ostii T. Hong & J. X. Zhang var. rockii (Stern) Pei & Hong peptide according to claim 2, wherein: In the composite enzyme hydrolysis and enzyme inactivation, add acid protease and flavor protease to the Danfeng peony flower protein solution that has undergone ultrasonic pretreatment, adjust to pH = 5.0, and then carry out dynamic circulation reaction at 55 °C. The degree of hydrolysis reaches 35%.

5. The anti-aging composition containing Paeonia ostii flower peptide according to claim 2, wherein: In step S13, first remove the unhydrolyzed protein through a 100 kDa ultrafiltration membrane, then enrich the active peptide segments through a 10 kDa nanofiltration membrane, and finally concentrate to a solid content of ≥25% through a reverse osmosis membrane.

6. The anti-aging composition containing Paeonia ostii T. Hong & J. X. Zhang flower peptide according to claim 1, wherein: The preparation method of the compound plant flavonoids is as follows: S21. Sub - critical water extraction. Crush Galinsoga parviflora, Taraxacum mongolicum, and Perilla frutescens leaves to obtain a compound plant powder. Then add sub - critical water at a ratio of 40:1 to the compound plant powder, maintain at a temperature of 120 °C ± 5 and a pressure of 3 MPa for 15 minutes, and then add 0.1% β - cyclodextrin to encapsulate the thermosensitive components to obtain flavonoids. S22. Macroporous resin purification. Select AB - 8 type resin for dynamic adsorption, with a sample loading flow rate of 2 BV / h and a sample loading concentration of 1.2 mg flavonoids / mL. The adsorption rate is > 95%. Then, through step - gradient elution, remove polysaccharides and pigment impurities to finally obtain the compound plant flavonoids.

7. The anti-aging composition containing Paeonia ostii Feng Dan peptide according to claim 6, wherein: The mixing ratio of the crushed Galinsoga parviflora, Taraxacum mongolicum, and Perilla frutescens leaves is 5:3:2, and they are ultra - micro - crushed to D90 ≤ 50 μm.

8. The anti-aging composition containing Paeonia ostii flower peptide according to claim 1, characterized in that: The step - gradient elution includes pretreatment elution, main elution, and fine elution. The pretreatment elution is carried out in a 30% ethanol solution at a flow rate of 3 BV / h to remove polysaccharides. The main elution is carried out in a 70% ethanol solution at a flow rate of 1.5 BV / h to remove quercetin / luteolin. The fine elution is carried out in a 90% ethanol solution at a flow rate of 2 BV / h to remove apigenin.

9. A method for preparing an anti-aging composition containing Paeonia ostii T. Hong & J. X. Zhang flower peptide according to any one of claims 1-8, characterized in that: Take the Danfeng peony flower peptide, compound plant flavonoids, chitosan oligosaccharide, sodium ascorbyl phosphate, and microencapsulated β - glucan by mass parts and mix them evenly to obtain.

10. Use of the anti - aging composition containing Danfeng peony flower peptide according to claims 1 - 8 in the preparation of anti - aging products.

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