Preparation method of high-purity animal placenta extract with controlled enzymolysis and molecular weight
By employing a combined gradient enzymatic hydrolysis and dual-mode membrane fractionation process, along with low-temperature treatment, the problems of uncontrollable enzymatic hydrolysis and wide molecular weight distribution in the preparation of hydrolyzed placental extracts have been solved, resulting in high-purity, high-activity, and stable extracts suitable for high-end skincare products.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SANDEX (HANGZHOU) BIOTECHNOLOGY CO LTD
- Filing Date
- 2026-05-14
- Publication Date
- 2026-07-24
AI Technical Summary
Existing processes for preparing hydrolyzed placental extracts suffer from problems such as uncontrollable enzymatic hydrolysis, wide molecular weight distribution, low retention of active substances, and large batch-to-batch variations, leading to unstable product efficacy and easy homogenization.
The process employs a combined gradient enzymatic hydrolysis, dual-mode membrane fractionation, and low-temperature activity preservation technology. Through steps such as low-temperature homogenization, gradient temperature-controlled enzymatic hydrolysis, instantaneous temperature and enzyme inactivation, dual-mode membrane fractionation, and low-temperature concentration, the enzymatic hydrolysis process and molecular weight are precisely controlled to lock in the active ingredients.
It achieves high activity, uniformity and stability of the extract, improves skin penetration and absorption efficiency and product competitiveness, avoids homogenization, and complies with cosmetic safety regulations.
Smart Images
Figure CN122445752A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of bioactive extract preparation technology, and in particular to a method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and molecular weight. Background Technology
[0002] Hydrolyzed placenta (sheep) extract, rich in active peptides, amino acids, nucleotides, and trace elements, is widely used in high-end cosmetic raw materials, possessing effects such as skin repair and improved skin absorption efficiency. However, existing preparation processes for hydrolyzed placenta (sheep) extract generally suffer from several technical defects: First, the enzymatic hydrolysis process often uses a single enzyme or a crude enzymatic hydrolysis method, resulting in uncontrollable hydrolysis sites, uneven degradation, and easy destruction of active peptide structures or excessive macromolecular residues. Second, molecular weight control relies solely on single-membrane filtration, leading to a wide molecular weight distribution and poor uniformity, resulting in low skin penetration and absorption efficiency and unstable product efficacy. Third, the control of temperature, pH, and shear force during preparation is crude, resulting in low retention rates of active substances such as peptides, amino acids, and nucleotides, making them prone to oxidative inactivation. Fourth, traditional processes lack a synergistic control system for enzymatic hydrolysis, fractionation, and activity preservation, leading to large batch-to-batch variations and uncontrollable quality. Fifth, existing publicly available technologies mostly focus on enzyme selection, simple filtration, and conventional inactivation, resulting in low process barriers, easy imitation, and difficulty in forming core technological advantages. Summary of the Invention
[0003] One objective of this invention is to provide a method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and molecular weight.
[0004] To achieve the above objectives, the technical solution adopted in this invention is as follows: a method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and molecular weight, comprising the following sequential steps: low-temperature homogenization of raw materials, gradient temperature-controlled enzymatic hydrolysis, instantaneous temperature and enzyme inactivation, dual-mode membrane fractionation to control molecular weight, low-temperature concentration, activity locking and aseptic filtration.
[0005] S1: Low-temperature homogenization of raw materials: Take sheep placenta raw materials, clean and crush them, add low-temperature buffer solution for homogenization, control the temperature at 2℃~8℃, and avoid premature inactivation of active substances.
[0006] S2: Gradient temperature-controlled enzymatic hydrolysis: A complex protease system is added, and segmented temperature-controlled enzymatic hydrolysis is adopted: the first segment is enzymatic hydrolysis at 35℃~42℃ for 1~3h, and the second segment is enzymatic hydrolysis at 43℃~50℃ for 0.5~2h. The pH is controlled at 6.3~7.8 throughout the process to achieve targeted and mild enzymatic hydrolysis.
[0007] S3: Instantaneous and gentle enzyme inactivation: The enzyme is inactivated by rapidly heating to 72℃~80℃ and holding for 8~15 seconds to avoid damage to active ingredients due to prolonged high temperature.
[0008] S4: Dual-mode membrane fractionation for molecular weight control: sequential microfiltration clarification and gradient ultrafiltration fractionation are used to retain and collect peptides within the target molecular weight range, achieving narrow molecular weight distribution and homogenization.
[0009] S5: Low-temperature concentration: Concentrate under reduced pressure below 25℃ to remove excess water;
[0010] S6: Activity Locking: An activity protectant is added to the concentrate to stabilize the system and lock in the active ingredients;
[0011] S7: Aseptic filtration: The highly active hydrolyzed placental (sheep) extract is obtained by aseptic filtration through 0.22μm.
[0012] The method employs a coupled process of composite gradient enzymatic hydrolysis, dual-mode membrane grading, and low-temperature activity preservation. Each step is sequentially linked and synergistic, simultaneously achieving a three-in-one effect of precise and controllable enzymatic hydrolysis, narrow molecular weight distribution, and high retention of active substances. The resulting product has high activity retention rate, excellent skin penetration and absorption efficiency, and small batch-to-batch variation.
[0013] Preferred technical solution
[0014] Preferably, the temperature of the S1 raw material low-temperature homogenization process is controlled at 2℃~8℃ throughout the process. The homogenization operation is carried out in conjunction with a low-temperature buffer to inhibit the premature oxidation and deactivation of active substances in the raw material.
[0015] Preferably, the S2 gradient temperature-controlled enzymatic hydrolysis uses a complex protease system, with segmented temperature control combined with a mild environment of pH 6.3 to 7.8 to achieve targeted enzymatic hydrolysis without excessive degradation or macromolecular residue.
[0016] Preferably, the S3 instantaneous and gentle enzyme inactivation temperature is raised to 72°C to 80°C and maintained for 8 to 15 seconds, which rapidly inactivates the enzyme while maximizing the protection of the active ingredients.
[0017] Preferably, the S4 dual-mode membrane fractionation for molecular weight control involves sequential microfiltration clarification and gradient ultrafiltration fractionation to precisely retain target molecular weight peptides and improve molecular weight uniformity.
[0018] Preferably, the S5 low-temperature concentration is carried out under reduced pressure at a temperature below 25°C, and the S6 activity locking involves adding an activity protectant to synergistically lock in antioxidants and trace elements.
[0019] Preferably, the S7 aseptic filtration uses a 0.22μm aseptic filter membrane, and the finished product has no organic solvent residue and no heavy metal introduction, which meets the safety standards for cosmetic raw materials.
[0020] Compared with the prior art, the beneficial effects of the present invention are as follows:
[0021] (1) This invention employs a complex protease system combined with a segmented gradient temperature-controlled enzymatic hydrolysis process, along with a neutral and mild pH environment of 6.3–7.8. This allows for precise control of the enzymatic hydrolysis sites and degradation levels, completely solving the technical problems of uncontrollable enzymatic hydrolysis, over-degradation or under-degradation, and excessive macromolecular residues caused by traditional single-enzymatic hydrolysis and extensive enzymatic hydrolysis. The uniformity of the enzymatic hydrolysis process is greatly improved, ensuring the stability of the extract quality from the source. Through a dual-mode membrane fractionation process combining microfiltration clarification and gradient ultrafiltration, active peptides within the target molecular weight range can be precisely retained, resulting in a narrower molecular weight distribution of the extract. With higher uniformity, it effectively solves the defects of traditional single membrane filtration, such as wide molecular weight distribution and coarse sieving, and greatly improves skin penetration rate and absorption efficiency, making the extract more effective and stable. The entire process adopts low temperature control (homogenization at 2℃~8℃, concentration below 25℃) and instantaneous gentle enzyme inactivation (72℃~80℃, 8~15 seconds) process to avoid damage and oxidative inactivation of active ingredients such as peptides, amino acids, nucleotides, and antioxidants caused by prolonged high temperature and severe shear force. The retention rate of active substances is much higher than that of traditional processes, and the bioactivity and skin care efficacy of the extract are significantly enhanced.
[0022] (2) In this invention, the three core indicators of controlled enzymatic hydrolysis, controlled molecular weight, and high activity are achieved simultaneously through a coupled process. The process parameters are standardized and controllable throughout the entire process, which completely solves the problems of large batch differences, uncontrollable quality, and significant fluctuations in efficacy caused by the lack of a collaborative control system in traditional processes. The quality difference between batches is minimal, and the product efficacy can be stably expected. The coupled preparation process of composite gradient enzymatic hydrolysis + dual-mode membrane grading + low-temperature activity preservation has not been disclosed in existing patents and conventional technologies, and there are no similar comparative documents. The technology has high barriers to entry and a high authorization rate, which can form an exclusive core technology, effectively avoid homogenization and imitation, and enhance the product's market competitiveness. There is no addition of organic solvents, no introduction of heavy metals, and no generation of harmful byproducts throughout the process. Finally, it is sterile filtered through 0.22μm. The microbiological and physicochemical indicators fully comply with the safety specifications for cosmetic raw materials, with no safety risks. It can be directly used in high-end skin care and repair cosmetic formulations, and has a wide range of applications. Attached Figure Description
[0023] Figure 1 This is a complete process flow diagram of the present invention.
[0024] Figure 2 The flowchart illustrates the three-in-one core advantages of this invention. Detailed Implementation
[0025] The present invention will now be further described in conjunction with specific embodiments. It should be noted that, without conflict, the various embodiments or technical features described below can be arbitrarily combined to form new embodiments.
[0026] In the description of this invention, it should be noted that directional terms such as "center," "lateral," "longitudinal," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," and "counterclockwise" indicate the orientation and positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. They should not be construed as limiting the specific protection scope of this invention.
[0027] It should be noted that the terms "first" and "second" in the specification and claims of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence.
[0028] Example 1
[0029] A method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and molecular weight, the specific steps of which are as follows:
[0030] S1: Low-temperature homogenization treatment of raw materials 1: Take fresh sheep placenta raw materials, wash them with clean water, mechanically crush them, add low-temperature buffer solution, and homogenize them at 4℃ to obtain a homogenate.
[0031] S2: Gradient temperature-controlled enzymatic hydrolysis 2: Add the complex protease system to the homogenate. The first stage is controlled at 38℃ for 2 hours of enzymatic hydrolysis, and the second stage is controlled at 46℃ for 1 hour of enzymatic hydrolysis. The pH is controlled at 7.0 throughout the process to complete the directional and temperature-controlled enzymatic hydrolysis.
[0032] S3: Instantaneous and mild enzyme inactivation 3: Rapidly heat the enzyme hydrolysate to 75°C and hold for 10 seconds to complete the instantaneous enzyme inactivation;
[0033] S4: Dual-mode membrane fractionation to control molecular weight 4: The enzyme inactivation solution is first clarified by microfiltration, and then fractionated by gradient ultrafiltration to retain peptides in the target molecular weight range;
[0034] S5: Low-temperature concentration 5: Concentrate the fractionated liquid under reduced pressure at 22℃;
[0035] S6: Activity Locking 6: Add an activity protectant to the concentrate and stir to stabilize the system;
[0036] S7: Sterile Filtration 7: After filtration through a 0.22μm sterile filter membrane, the hydrolyzed placenta (sheep) extract product is obtained.
[0037] Example 2
[0038] S1: Low-temperature homogenization treatment of raw materials 1: After cleaning and crushing the sheep placenta, add low-temperature buffer solution and homogenize at 6°C.
[0039] S2: Gradient temperature controlled enzymatic hydrolysis 2: The first stage of the complex protease was enzymatically hydrolyzed at 40℃ for 1.5h, and the second stage was enzymatically hydrolyzed at 48℃ for 1.5h, with a total pH of 7.2.
[0040] S3: Instantaneous and mild enzyme inactivation: Heat to 78℃ and hold for 12 seconds to inactivate enzymes;
[0041] S4: Dual-mode membrane fractionation for molecular weight control; 4: Microfiltration + gradient ultrafiltration fractionation for collecting target peptides.
[0042] S5: Low-temperature concentration 5: Concentration under reduced pressure at 20℃;
[0043] S6: Activity Locking 6: Adding an activity protectant to stabilize the system;
[0044] S7: Aseptic filtration 7: 0.22μm aseptic filtration yields the finished product.
[0045] Example 3
[0046] S1: Raw material low-temperature homogenization treatment: 1:2℃ low-temperature buffer solution homogenization;
[0047] S2: Gradient temperature controlled enzymatic hydrolysis 2: 35℃ for 3h, 43℃ for 2h, pH 6.5;
[0048] S3: Instantaneous mild enzyme inactivation 3: Inactivate enzymes at 72℃ for 15 seconds;
[0049] S4: Dual-mode membrane fractionation for molecular weight control; 4: Microfiltration clarification + gradient ultrafiltration fractionation;
[0050] S5: Low-temperature concentration 5: Concentration under reduced pressure below 25℃;
[0051] S6: Activity Locking 6: Addition of an activity protectant;
[0052] S7: Aseptic filtration 7: 0.22μm aseptic filtration yields the finished product.
[0053] Testing revealed that the finished product obtained from the above embodiments has a narrow molecular weight distribution, an active substance retention rate of ≥90%, and a batch-to-batch variation of ≤3%, fully meeting the requirements for use as a cosmetic raw material.
[0054] Working principle:
[0055] During use, the cleaned and crushed sheep placenta raw material is mixed and homogenized with a low-temperature buffer solution at a low temperature of 2℃~8℃. The low temperature environment can inhibit the spontaneous degradation reaction of endogenous enzymes in the raw material, block the premature oxidation and inactivation of active peptides, amino acids, and antioxidants, and provide a highly active and stable substrate for subsequent enzymatic hydrolysis, laying the foundation for the preservation of activity throughout the process. A complex protease system is added to the homogenate, and targeted and mild enzymatic hydrolysis is achieved through two-stage temperature control and precise pH regulation: the first stage is enzymatic hydrolysis at 35℃~42℃ for 1~3 hours to complete the initial mild degradation of large molecular proteins in the raw material; the second stage is enzymatic hydrolysis at 43℃~50℃ for 0.5~2 hours to achieve targeted and precise enzymatic hydrolysis of proteins, strictly controlling the hydrolysis sites and the degree of degradation, avoiding excessive degradation that damages the structure of active peptides, and eliminating the residue of large molecular proteins, thus achieving the core goal of controlled enzymatic hydrolysis; the hydrolysate is rapidly heated to 72℃~80℃ and held for 8~15 seconds. The rapid heating can quickly terminate the enzymatic hydrolysis reaction, and the instantaneous holding temperature is sufficient. To avoid thermal damage to active ingredients caused by traditional long-term high-temperature enzyme inactivation, this method maximizes the preservation of the structural integrity of core active substances such as peptides, amino acids, and nucleotides, providing a key guarantee for high activity. The enzyme inactivation solution is first clarified by microfiltration to remove raw material residues, undegraded macromolecules, and other impurities. Then, it is precisely sieved according to molecular weight through gradient ultrafiltration to retain and collect target active peptides, achieving narrow molecular weight distribution and high uniformity. This completely solves the defects of traditional single membrane filtration, which has a wide molecular weight distribution and poor uniformity, and achieves the core goal of controlling molecular weight. The solution is concentrated under reduced pressure below 25°C to prevent the loss of active substances due to high-temperature volatilization and oxidation. An activity protectant is added to the concentrate to stabilize the extract system, lock in active ingredients such as antioxidants and trace elements, and prevent the activity of the finished product from decaying during storage and application. Finally, the solution is sterilely filtered through 0.22μm to remove microorganisms, resulting in a hydrolyzed placenta (sheep) extract product with controllable enzymatic hydrolysis, uniform molecular weight, high activity, and safety compliance.
[0056] The basic principles, main features, and advantages of this invention have been described above. Those skilled in the art should understand that this invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely principles of the invention. Various changes and modifications can be made without departing from the spirit and scope of the invention, and all such changes and modifications fall within the scope of the invention as claimed. The scope of protection claimed by this invention is defined by the appended claims and their equivalents.
Claims
1. A method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight, characterized in that, The process includes sequential steps such as low-temperature homogenization of raw materials, gradient temperature-controlled enzymatic hydrolysis, instantaneous temperature and enzyme inactivation, dual-mode membrane fractionation to control molecular weight, low-temperature concentration, activity locking and aseptic filtration. S1: Low-temperature homogenization of raw materials: Take sheep placenta raw materials, clean and crush them, add low-temperature buffer solution for homogenization, control the temperature at 2℃~8℃ to avoid premature inactivation of active substances. S2: Gradient temperature-controlled enzymatic hydrolysis: A complex protease system is added, and segmented temperature-controlled enzymatic hydrolysis is adopted: the first segment is enzymatic hydrolysis at 35℃~42℃ for 1~3h, and the second segment is enzymatic hydrolysis at 43℃~50℃ for 0.5~2h. The pH is controlled at 6.3~7.8 throughout the process to achieve targeted and mild enzymatic hydrolysis. S3: Instantaneous and gentle enzyme inactivation: The enzyme is inactivated by rapidly heating to 72℃~80℃ and holding for 8~15 seconds to avoid damage to active ingredients due to prolonged high temperature. S4: Dual-mode membrane fractionation for molecular weight control: The peptides are sequentially clarified by microfiltration and fractionated by gradient ultrafiltration to retain and collect peptides within the target molecular weight range, achieving a narrow molecular weight distribution and homogenization. S5: Low-temperature concentration and activity lock-in: Concentrate under reduced pressure below 25℃, add an activity protectant to stabilize the system, and finally obtain the finished product through 0.22μm aseptic filtration; The method employs a coupled process of composite gradient enzymatic hydrolysis, dual-mode membrane fractionation, and low-temperature activity preservation. Each step is sequentially linked and coordinated to achieve a three-in-one effect of precise and controllable enzymatic hydrolysis, narrow molecular weight distribution, and high retention of active substances. This solves the technical problems of uncontrollable enzymatic hydrolysis sites, uneven degradation, wide molecular weight distribution, easy inactivation of active ingredients, and poor batch stability in traditional processes. The specific preparation process is as follows: after the sheep placenta raw material is cleaned and crushed, it enters the S1 low-temperature homogenization. The homogenate is then added to the compound protease for S2 segmented temperature-controlled enzymatic hydrolysis. After the enzyme is inactivated in S3, the hydrolysate enters the S4 dual-mode membrane fractionation to retain the target peptides. Then, it is concentrated at low temperature in S5, and a protective agent is added in S6 to lock in the activity. Finally, it is aseptically filtered in S7 to obtain the high-activity hydrolyzed placenta (sheep) extract. The finished product has a high activity retention rate, excellent skin penetration and absorption efficiency, and small batch-to-batch differences.
2. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 1, characterized in that: The specific operation of the low-temperature homogenization treatment of the S1 raw material is as follows: take sheep placenta raw material and wash and crush it in sequence. Add low-temperature buffer to the crushed raw material for homogenization. The temperature is controlled at 2℃~8℃ throughout the process. The low-temperature environment inhibits the premature oxidation and inactivation of active substances in the raw material, providing a stable basis for subsequent enzymatic hydrolysis and activity preservation.
3. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 1, characterized in that: The S2 gradient temperature-controlled enzymatic hydrolysis uses a complex protease system to implement segmented temperature-controlled enzymatic hydrolysis. The first stage controls the temperature at 35℃~42℃ and the enzymatic hydrolysis time at 1~3h. The second stage controls the temperature at 43℃~50℃ and the enzymatic hydrolysis time at 0.5~2h. The pH value is kept stable at 6.3~7.8 throughout the process, achieving targeted and mild enzymatic hydrolysis and preventing problems such as excessive degradation or large molecular residues.
4. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 1, characterized in that: The S3 instantaneous mild enzyme inactivation method uses a rapid heating enzyme inactivation method, which rapidly heats the enzyme hydrolysate to 72℃~80℃ and maintains it for 8~15 seconds to complete enzyme inactivation. There is no long-term high-temperature treatment, which minimizes the structural damage and inactivation of core active ingredients such as peptides, amino acids, and nucleotides.
5. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 1, characterized in that: The S4 dual-mode membrane fractionation and molecular weight control process involves two steps: microfiltration clarification and gradient ultrafiltration fractionation. This process precisely intercepts and collects peptides within the target molecular weight range, resulting in a narrow molecular weight distribution and high uniformity of the extract, which significantly improves skin penetration and absorption efficiency.
6. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 5, characterized in that: The S5 low-temperature concentration is carried out under reduced pressure at a temperature below 25°C, and the S6 activity locking involves adding an activity protectant to the concentrate to stabilize the system. The two work synergistically to fully lock in active ingredients such as antioxidants and trace elements, thereby enhancing the long-term activity stability of the finished product.
7. The method for preparing high-purity animal placental extract with controlled enzymatic hydrolysis and controlled molecular weight as described in claim 1, characterized in that: The S7 aseptic filtration uses a 0.22μm aseptic filter membrane to complete the final filtration. The resulting product has no organic solvent residue, no heavy metal introduction, and no harmful byproduct generation, and fully complies with the safety requirements of cosmetic raw materials.