Refining process and application of hydrolyzed sponge

Abstract

The invention relates to the technical field of hydrolyzed sponge, and discloses a hydrolyzed sponge refining process and application, the process comprises the following steps: raw material pretreatment, natural sponge raw material selection, artificial grading and impurity removal, direct hydrolysis after impurity removal, dilute acid hydrolysis, and putting the pretreated raw material into an acid-resistant reaction container, the preparation method comprises the following steps: adding a dilute acid solution with the mass fraction of 1%-5%, carrying out a stepped hydrolysis reaction at the temperature of 50-90 DEG C for 6-12 hours, filtering and cleaning, and carrying out a hydrolysis process with sectional temperature control and auxiliary treatment with a surfactant during refining processing of the hydrolyzed sponge spicule, so as to obtain the hydrolyzed sponge spicule. According to the method, the microstructure integrity and biological activity of the sponge spicule can be effectively reserved, meanwhile, proper hydrolysis parameters are set according to sponge raw materials from different sources, the controllability of the refining process of the sponge spicule is guaranteed, the purity and quality stability of the product are improved, and a material foundation is laid for subsequent application.

Description

Technical Field

[0001] This invention relates to the field of hydrolyzed sponge technology, specifically to the refining process and application of hydrolyzed sponges. Background Technology

[0002] Hydrolyzed sponge is an extremely tiny needle-like bone extracted from siliceous sponge. It is invisible to the naked eye, but appears as a needle with rounded or pointed ends under a microscope, hence the name sponge microneedle.

[0003] Currently, due to the diversity of sponge raw material sources and the complexity of its own structure, the traditional acid-base treatment process used in the hydrolysis and refining of sponge spicules cannot accurately control the impact of the degree of hydrolysis on the spicule structure when removing organic impurities. When excessive or insufficient hydrolysis occurs during the process, it will cause damage to the spicule structure or impurity residue, and the quality stability of the product cannot be guaranteed.

[0004] Therefore, a refining process and application of hydrolyzed sponge bone needles are proposed to solve the above problems. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a refining process and application for hydrolyzed sponges, solving the problem mentioned in the background art of bone needle structure damage or impurity residue, which makes it impossible to guarantee the quality stability of the product.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a refining process and application of hydrolyzed sponges, comprising the following steps: S10: Raw material pretreatment, select natural sponge raw materials, manually sort and remove impurities, and directly enter the hydrolysis process after removing impurities; S20: dilute acid hydrolysis. The pretreated raw materials are put into an acid-resistant reaction vessel, and a dilute acid solution with a mass fraction of 1%-5% is added. The step hydrolysis reaction is carried out at a temperature of 50-90℃, and the total reaction time is 6-12 hours. S30: Filtration and cleaning. Use an 80-350 mesh filter to filter the hydrolysis products in stages, collect the sponge needles, and rinse with deionized water until neutral. S40: Alkali neutralization. Mix the cleaned bone needles with an alkali solution of 0.5%-2% by mass, stir slowly for 0.5-1 hour, and let stand. S50: pH adjustment, detects the pH value of the bone needle and adjusts it to the range of 6.5-7.5; S60: Drying treatment, drying the neutralized bone needles at 70-75℃ to constant weight; S70: Grading and screening, using a vibrating sieve to classify the dried bone needles by particle size, and selecting bone needle finished products with uniform particle size. The dilute acid hydrolysis adopts a segmented temperature control: first, pretreatment at 50-60℃ for 1-2 hours, then heating to 70-80℃ for main hydrolysis for 4-8 hours, and finally posttreatment at 85-90℃ for 1-2 hours. The hydrolysis process involves the addition of a surfactant as an auxiliary treatment. The surfactant is selected from either polysorbate-80 or alkyl glycoside, and the amount added is 0.1%-0.5% of the raw material mass.

[0007] Preferably, in step S20, the dilute acid solution is selected from hydrochloric acid or sulfuric acid, with a mass fraction preferably of 2%-4%. The hydrolysis reaction vessel is selected from enamel-lined reaction tanks or fiberglass reaction kettles. The stirring speed is controlled at 50-100 r / min, and an intermittent stirring method is adopted. The stirring is carried out for 30 minutes, then the stirring is stopped and the mixture is allowed to stand for 6 hours, and then stirred again for 30 minutes.

[0008] Preferably, in step S30, the filter mesh size is preferably 100-300 mesh, the rinsing water is deionized water with a resistivity of not less than 15 MΩ·cm, and the standard for rinsing to neutral is that the pH value of the rinsing solution reaches 6.5-7.5.

[0009] Preferably, in step S40, the alkaline solution is selected from sodium hydroxide or potassium hydroxide solution, the mass fraction is preferably 1%-1.5%, the neutralization temperature is controlled at 25-35℃, and the stirring speed is controlled at 30-50 r / min.

[0010] Preferably, in step S60, the drying temperature is 72-74℃, the drying time is 4-6 hours, and the drying equipment is a vacuum drying oven or a hot air circulating drying oven. The standard for drying to constant weight is that the difference in mass between two consecutive weighings does not exceed 0.5%.

[0011] Preferably, it includes the following steps: A10: Mix hydrolyzed sponge bone needles with a special solvent at a mass ratio of 1:1 to 1:3 to prepare a uniform paste. A20: Apply the paste to the skin surface and treat it using a specific transdermal massage technique for 3-5 minutes; A30: After removing surface powder, use the active ingredient preparation for subsequent care; A40: Establish a system for verifying the efficacy of sponge bone needles, including in vitro permeability testing and in vivo efficacy assessment; The specially prepared solvent contains glycerol, propylene glycol and deionized water in a mass ratio of 1:1:2 to 1:2:3. The massage technique uses a reverse pore spiral lifting method, with pressure controlled within the range of 20-50 grams. The active ingredient formulation contains a functional ingredient with a molecular weight of 500-5000 Da and a mass fraction of 0.5%-5%.

[0012] Preferably, in step A10, the special solvent further contains 0.1%-0.5% of a preservative and 0.01%-0.05% of an antioxidant. The preservative is selected from phenoxyethanol or potassium sorbate, and the antioxidant is selected from vitamin E or tert-butylhydroquinone.

[0013] Preferably, in step A20, the massage technique specifically includes: First, apply 20-30 grams of pressure and massage in a circular motion for 1-2 minutes. Then, apply 30-50 grams of pressure and massage against the pores for 2-3 minutes. The massage frequency is 60-80 times per minute.

[0014] Preferably, in step A40, the in vitro permeability test uses the Franz diffusion cell method: Using pig ear skin or artificial skin as a permeability barrier, the test temperature is 32-37℃, and the sampling time points are 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours. The detection method is HPLC or fluorescence spectrophotometry.

[0015] Preferably, in step A40, the in vivo efficacy assessment includes: Animal models or human volunteers were used, and the evaluation indicators included skin hydration, transepidermal water loss, skin elasticity, melanin index, and hemoglobin index. The testing period was 1-4 weeks, and the data analysis used statistical methods with a p-value of less than 0.05.

[0016] Beneficial effects Compared with the prior art, the present invention provides a refining process and application for hydrolyzed sponges, which has the following beneficial effects: 1. In this invention, during the refining process of hydrolyzed sponge spicules, a segmented temperature-controlled hydrolysis process and surfactant-assisted treatment are adopted to effectively preserve the microstructural integrity and bioactivity of the sponge spicules. At the same time, appropriate hydrolysis parameters are set for sponge raw materials from different sources to ensure the controllability of the sponge spicule refining process. This solves the problems of spicule structure damage and impurity residue caused by improper hydrolysis in traditional processes, improves the purity and quality stability of the product, and lays a material foundation for subsequent applications.

[0017] 2. In this invention, during the purification process of sponge bone needles, a graded filtration and screening system is established to achieve precise grading based on the particle size and morphological characteristics of the bone needles. This allows for effective control of product uniformity and enables precise dosage control during application, avoiding differences in application effects due to poor product uniformity. This further improves the applicability and reliability of sponge bone needles in various application scenarios.

[0018] 3. In the practical application of sponge bone needles, by developing a special solvent formula and specific transdermal massage techniques, and establishing a complete in vivo efficacy evaluation system, customized application solutions can be achieved for different application scenarios. This effectively solves the problem of fluctuations in application effects caused by the lack of a dedicated delivery system and effect evaluation system in traditional applications, improves the efficiency of sponge bone needles in terms of functional component loading and transdermal delivery, and further expands its application effects and market value in high value-added fields. Attached Figure Description

[0019] Figure 1 This is a flowchart of the refining process for the hydrolyzed sponge of the present invention; Figure 2 This is a flowchart illustrating the application method of the refining process for hydrolyzed sponge according to the present invention. Detailed Implementation

[0020] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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 skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] Example 1: The refining process and application of this hydrolyzed sponge includes the following steps: S10: Raw material pretreatment, select natural sponge raw materials, manually sort and remove impurities, and directly enter the hydrolysis process after removing impurities; S20: Dilute acid hydrolysis. The pretreated raw material is put into an acid-resistant reaction vessel, and a 1% dilute acid solution is added. The step hydrolysis reaction is carried out at 50°C for a total reaction time of 6 hours. S30: Filtration and cleaning. Use an 80-mesh filter to filter the hydrolysis products in stages, collect the sponge needles, and rinse with deionized water until neutral. S40: Alkali neutralization: Mix the cleaned bone needles with an alkali solution with a mass fraction of 0.5%, stir slowly for 0.5 hours, and let stand. S50: pH adjustment, detects the pH value of the bone needle and adjusts it to the range of 6.5; S60: Drying treatment, the neutralized bone needles are dried to constant weight at 70°C; S70: Grading and screening, using a vibrating sieve to classify the dried bone needles by particle size, and selecting bone needle finished products with uniform particle size. The dilute acid hydrolysis process employs segmented temperature control: first, pretreatment at 50℃ for 1 hour, then heating to 70℃ for 4 hours of main hydrolysis, and finally posttreatment at 85℃ for 1 hour. A surfactant is added to assist the hydrolysis process. The surfactant is either polysorbate-80 or alkyl glycoside, and the amount added is 0.1% of the raw material mass. In step S20, the dilute acid solution is selected from hydrochloric acid or sulfuric acid, with a mass fraction preferably of 2%. The hydrolysis reaction vessel is selected from enamel-lined reaction tank or fiberglass reaction vessel. The stirring speed is controlled at 50 r / min, and an intermittent stirring method is adopted. Stirring is performed for 30 minutes, stirring is stopped and the mixture is allowed to stand for 6 hours, and then stirred again for 30 minutes. In step S30, the preferred mesh size of the filter screen is 100 mesh, the rinsing water is deionized water with a resistivity of not less than 15 MΩ·cm, and the standard for rinsing to neutral is that the pH value of the rinsing solution reaches 6.5. In step S40, the alkaline solution is selected from sodium hydroxide or potassium hydroxide solution, the mass fraction of which is preferably 1%, the neutralization temperature is controlled at 25℃, and the stirring speed is controlled at 30r / min. In step S60, the preferred drying temperature is 72℃, the drying time is 4 hours, and the drying equipment is a vacuum drying oven or a hot air circulating drying oven. The standard for drying to constant weight is that the difference between two consecutive weighings does not exceed 0.5%. Includes the following steps: A10: Hydrolyzed sponge bone needles are mixed with a special solvent at a mass ratio of 1:1 to prepare a uniform paste. A20: Apply the paste to the skin surface and treat it using a specific transdermal massage technique for 3 minutes; A30: After removing surface powder, use the active ingredient preparation for subsequent care; A40: Establish a system for verifying the efficacy of sponge bone needles, including in vitro permeability testing and in vivo efficacy assessment; The special solvent contains glycerin, propylene glycol and deionized water in a mass ratio of 1:1:2. The massage technique uses a reverse spiral lifting motion against the pores, with pressure controlled within 20 grams. The active ingredient formulation contains functional ingredients with a molecular weight of 500 Da, at a mass fraction of 0.5%. In step A10, the special solvent also contains 0.1% preservative and 0.01% antioxidant. The preservative is selected from phenoxyethanol or potassium sorbate, and the antioxidant is selected from vitamin E or tert-butylhydroquinone. In step A20, the massage techniques specifically include: First, massage in a circular motion with 20 grams of pressure for 1 minute, then massage against the pores with 30 grams of pressure for 2 minutes, at a frequency of 60 times per minute. In step A40, the in vitro permeability test uses the Franz diffusion cell method: Using pig ear skin or artificial skin as a permeation barrier, the test temperature was 32℃, and the sampling time points were 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours. The detection method was HPLC or fluorescence spectrophotometry. In step A40, the in vivo efficacy assessment includes: Animal models or human volunteers were used, and the evaluation indicators included skin hydration, transepidermal water loss, skin elasticity, melanin index, and hemoglobin index. The testing period was one week, and the data analysis used statistical methods with a p-value of less than 0.05.

[0022] Example 2: The refining process and application of this hydrolyzed sponge includes the following steps: S10: Raw material pretreatment, select natural sponge raw materials, manually sort and remove impurities, and directly enter the hydrolysis process after removing impurities; S20: Dilute acid hydrolysis. The pretreated raw material is put into an acid-resistant reaction vessel, and a 3% dilute acid solution is added. The step hydrolysis reaction is carried out at 70°C for a total reaction time of 9 hours. S30: Filtration and cleaning. Use a 250-mesh filter to filter the hydrolysis products in stages, collect the sponge needles, and rinse with deionized water until neutral. S40: Alkali neutralization: Mix the cleaned bone needles with an alkali solution of 1% by mass, stir slowly for 0.7 hours, and let stand. S50: pH adjustment, detects the pH value of the bone needle and adjusts it to the range of 7.0; S60: Drying treatment, the neutralized bone needles are dried at 72°C to constant weight; S70: Grading and screening, using a vibrating sieve to classify the dried bone needles by particle size, and selecting bone needle finished products with uniform particle size. The dilute acid hydrolysis process employs segmented temperature control: first, pretreatment at 50-60℃ for 1.5 hours, then heating to 75℃ for main hydrolysis for 6 hours, and finally posttreatment at 87℃ for 1.5 hours. A surfactant is added to assist the hydrolysis process. The surfactant is either polysorbate-80 or alkyl glycoside, and the amount added is 0.3% of the raw material mass. In step S20, the dilute acid solution is selected from hydrochloric acid or sulfuric acid, with a mass fraction preferably of 3%. The hydrolysis reaction vessel is selected from enamel-lined reaction tank or fiberglass reaction vessel. The stirring speed is controlled at 70 r / min. The stirring method is adopted, stirring for 30 minutes, stopping stirring and letting stand for 6 hours, and stirring again for 30 minutes. In step S30, the preferred filter mesh size is 200 mesh, the rinsing water is deionized water with a resistivity of not less than 15 MΩ·cm, and the standard for rinsing to neutral is that the pH value of the rinsing solution reaches 7.0. In step S40, the alkaline solution is selected from sodium hydroxide or potassium hydroxide solution, the mass fraction of which is preferably 1.2%, the neutralization temperature is controlled at 30℃, and the stirring speed is controlled at 40r / min; In step S60, the preferred drying temperature is 73℃, the drying time is 5 hours, and the drying equipment is a vacuum drying oven or a hot air circulating drying oven. The standard for drying to constant weight is that the difference in mass between two consecutive weighings does not exceed 0.5%. Includes the following steps: A10: Hydrolyzed sponge bone needles are mixed with a special solvent at a mass ratio of 1:2 to prepare a uniform paste. A20: Apply the paste to the skin surface and treat it using a specific transdermal massage technique for 4 minutes; A30: After removing surface powder, use the active ingredient preparation for subsequent care; A40: Establish a system for verifying the efficacy of sponge bone needles, including in vitro permeability testing and in vivo efficacy assessment; The specially formulated solvent contains glycerin, propylene glycol, and deionized water in a mass ratio of 1:1.5:2.5. The massage technique uses a reverse spiral lifting motion against the pores, with the pressure controlled within a range of 35 grams. The active ingredient formulation contains functional components with a molecular weight of 2500 Da, at a mass fraction of 3.5%. In step A10, the special solvent also contains 0.3% preservative and 0.03% antioxidant. The preservative is selected from phenoxyethanol or potassium sorbate, and the antioxidant is selected from vitamin E or tert-butylhydroquinone. In step A20, the massage techniques specifically include: First, apply 25 grams of pressure and massage in a circular motion for 1.5 minutes, then apply 40 grams of pressure and massage against the pores for 2.5 minutes, with a massage frequency of 70 times per minute; In step A40, the in vitro permeability test uses the Franz diffusion cell method: Using pig ear skin or artificial skin as a permeability barrier, the test temperature was 35℃, and the sampling time points were 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours. The detection method was HPLC or fluorescence spectrophotometry. In step A40, the in vivo efficacy assessment includes: Animal models or human volunteers were used, and the evaluation indicators included skin hydration, transepidermal water loss, skin elasticity, melanin index, and hemoglobin index. The testing period was 2 weeks, and the data analysis used statistical methods with a p-value of less than 0.05.

[0023] Example 3: The refining process and application of this hydrolyzed sponge includes the following steps: S10: Raw material pretreatment, select natural sponge raw materials, manually sort and remove impurities, and directly enter the hydrolysis process after removing impurities; S20: Dilute acid hydrolysis. The pretreated raw material is put into an acid-resistant reaction vessel, and a 5% dilute acid solution is added. The step hydrolysis reaction is carried out at 90℃ for a total reaction time of 12 hours. S30: Filtration and cleaning. Use a 350-mesh filter to filter the hydrolysis products in stages, collect the sponge needles, and rinse with deionized water until neutral. S40: Alkali neutralization. Mix the cleaned bone needles with an alkali solution of 2% by mass, stir slowly for 1 hour, and let stand. S50: pH adjustment, detects the pH value of the bone needle and adjusts it to within the range of 7.5; S60: Drying treatment, the neutralized bone needles are dried to constant weight at 75°C; S70: Grading and screening, using a vibrating sieve to classify the dried bone needles by particle size, and selecting bone needle finished products with uniform particle size. The dilute acid hydrolysis process employs segmented temperature control: first, pretreatment at 60℃ for 2 hours, then heating to 80℃ for main hydrolysis for 8 hours, and finally posttreatment at 90℃ for 2 hours. A surfactant is added to assist the hydrolysis process. The surfactant is either polysorbate-80 or alkyl glycoside, and the amount added is 0.5% of the raw material mass. In step S20, the dilute acid solution is selected from hydrochloric acid or sulfuric acid, with a mass fraction preferably of 4%. The hydrolysis reaction vessel is selected from enamel-lined reaction tank or fiberglass reaction vessel. The stirring speed is controlled at 100 r / min, and an intermittent stirring method is adopted. Stirring is performed for 30 minutes, stirring is stopped and the mixture is allowed to stand for 6 hours, and then stirred again for 30 minutes. In step S30, the preferred mesh size of the filter screen is 300 mesh, the rinsing water is deionized water with a resistivity of not less than 15 MΩ·cm, and the standard for rinsing to neutral is that the pH value of the rinsing solution reaches 7.5. In step S40, the alkaline solution is selected from sodium hydroxide or potassium hydroxide solution, the mass fraction of which is preferably 1.5%, the neutralization temperature is controlled at 35℃, and the stirring speed is controlled at 50r / min; In step S60, the drying temperature is preferably 74℃, the drying time is 6 hours, and the drying equipment is a vacuum drying oven or a hot air circulating drying oven. The standard for drying to constant weight is that the difference in mass between two consecutive weighings does not exceed 0.5%. Includes the following steps: A10: Hydrolyzed sponge bone needles are mixed with a special solvent at a mass ratio of 1:3 to prepare a uniform paste. A20: Apply the paste to the skin surface and treat it with a specific transdermal massage technique for 5 minutes; A30: After removing surface powder, use the active ingredient preparation for subsequent care; A40: Establish a system for verifying the efficacy of sponge bone needles, including in vitro permeability testing and in vivo efficacy assessment; The special solvent contains glycerin, propylene glycol and deionized water in a mass ratio of 1:2:3. The massage technique uses a reverse spiral lifting motion against the pores, with pressure controlled within 50 grams. The active ingredient formulation contains functional ingredients with a molecular weight of 5000 Da, comprising 5% by mass. In step A10, the special solvent also contains 0.5% preservative and 0.05% antioxidant. The preservative is selected from phenoxyethanol or potassium sorbate, and the antioxidant is selected from vitamin E or tert-butylhydroquinone. In step A20, the massage techniques specifically include: First, apply 30 grams of pressure and massage in a circular motion for 2 minutes, then apply 50 grams of pressure and massage against the pores for 3 minutes, with a massage frequency of 80 times per minute. In step A40, the in vitro permeability test uses the Franz diffusion cell method: Using pig ear skin or artificial skin as a permeability barrier, the test temperature was 37℃, and the sampling time points were 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours. The detection method was HPLC or fluorescence spectrophotometry. In step A40, the in vivo efficacy assessment includes: Using animal models or human volunteers, the evaluation indicators included skin hydration, transepidermal water loss, skin elasticity, melanin index, and hemoglobin index. The testing period was 4 weeks, and the data analysis used statistical methods with a p-value of less than 0.05.

[0024] Comparative Example 1 differs from Example 1 in that: this comparative example does not use stepped temperature control during the hydrolysis process, but instead uses a single temperature condition of constant 80°C for the hydrolysis reaction.

[0025] Comparative Example 2 differs from Example 2 in that: no surfactant was added during the purification process of this comparative example, and only conventional acid-base treatment was used.

[0026] Comparative Example 3 differs from Example 3 in that: this comparative example did not undergo a grading and screening process, and directly used the dried bone needle product.

[0027] Comparative Example 4 differs from Example 3 in that it does not use a special solvent but instead uses ordinary deionized water as the dispersion medium.

[0028] The performance of the hydrolyzed sponge bone needles prepared in Examples 1-3 and Comparative Examples 1-4 was tested. The test items and test methods are as follows: The integrity rate of bone spurs was tested by observing the surface morphology of bone spurs using a scanning electron microscope, and the proportion of structurally intact bone spurs out of 100 bone spurs was counted to calculate the integrity rate percentage. Particle size distribution test: The particle size distribution of bone needle samples was measured using a laser particle size analyzer, and the D10, D50, and D90 values ​​were recorded. pH stability test: bone needle samples were soaked in deionized water for 24 hours and the range of pH change of the soaking solution was measured. Transdermal absorption rate was tested using the Franz diffusion cell method, with FITC-OVA as the model drug. The cumulative permeation was calculated over 6 hours at a test temperature of 32±1℃. To assess the rate of skin improvement, a 4-week clinical trial was conducted with 30 volunteers to measure the rate of change in skin hydration and transepidermal water loss.

[0029] The test data of the hydrolyzed sponge spicules prepared in Examples 1-3 and Comparative Examples 1-4 are recorded in the table below:

[0030] Comparison and analysis of the data in the table show that the hydrolyzed sponge spicules prepared using the processes in Examples 1-3 exhibit significantly superior performance compared to the products prepared using the processes in Comparative Examples 1-4. This indicates that during the refining of hydrolyzed sponge spicules, the use of segmented temperature-controlled hydrolysis processes and surfactant-assisted treatment can effectively preserve the microstructural integrity and bioactivity of the sponge spicules. Furthermore, setting appropriate hydrolysis parameters for sponge raw materials from different sources ensures the controllability of the sponge spicule refining process, solving the problems of spicule structural damage and impurity residue caused by improper hydrolysis in traditional processes. This improves the purity and quality stability of the product, laying a material foundation for subsequent applications. In the refining and purification process of sponge spicules, a graded filtration and screening system is established. Precise grading based on the particle size and morphological characteristics of the bone needles enables effective control of product uniformity. Simultaneously, precise dosage control during application avoids variations in application effects caused by poor product uniformity, further enhancing the applicability and reliability of sponge bone needles in various application scenarios. In practical applications, by developing specialized solvent formulations and specific transdermal massage techniques, and establishing a complete in vivo efficacy evaluation system, customized application solutions can be implemented for different scenarios. This effectively solves the problem of fluctuating application effects caused by the lack of dedicated delivery systems and efficacy evaluation systems in traditional applications, improving the efficiency of sponge bone needles in functional ingredient loading and transdermal delivery, and further expanding their application effects and market value in high-value-added fields.

[0031] By comparing and analyzing the relevant data in the table, it can be seen that the hydrolyzed sponge bone needles prepared by the process of the present invention not only have high structural integrity, controllability and reliability, but also indicate that the hydrolyzed sponge bone needle refining process and application provided by the present invention have a broader market prospect and are more suitable for promotion.

[0032] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0033] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A refining process for hydrolyzed sponges, characterized in that, Includes the following steps: S10: Raw material pretreatment, select natural sponge raw materials, manually sort and remove impurities, and directly enter the hydrolysis process after removing impurities; S20: dilute acid hydrolysis. The pretreated raw materials are put into an acid-resistant reaction vessel, and a dilute acid solution with a mass fraction of 1%-5% is added. The step hydrolysis reaction is carried out at a temperature of 50-90℃, and the total reaction time is 6-12 hours. S30: Filtration and cleaning. Use an 80-350 mesh filter to filter the hydrolysis products in stages, collect the sponge needles, and rinse with deionized water until neutral. S40: Alkali neutralization. Mix the cleaned bone needles with an alkali solution of 0.5%-2% by mass, stir slowly for 0.5-1 hour, and let stand. S50: pH adjustment, detects the pH value of the bone needle and adjusts it to the range of 6.5-7.5; S60: Drying treatment, drying the neutralized bone needles at 70-75℃ to constant weight; S70: Grading and screening, using a vibrating sieve to classify the dried bone needles by particle size, and selecting bone needle finished products with uniform particle size. The dilute acid hydrolysis adopts a segmented temperature control: first, pretreatment at 50-60℃ for 1-2 hours, then heating to 70-80℃ for main hydrolysis for 4-8 hours, and finally posttreatment at 85-90℃ for 1-2 hours. In the hydrolysis process, a surfactant is specially added to assist the treatment. The surfactant is selected from polysorbate-80 or alkyl glycoside, and the amount added is 0.1%-0.5% of the raw material mass.

2. The refining process for hydrolyzed sponge according to claim 1, characterized in that: In step S20, the dilute acid solution is selected from hydrochloric acid or sulfuric acid, with a mass fraction preferably of 2%-4%. The hydrolysis reaction vessel is selected from enamel-lined reaction tanks or fiberglass reaction kettles. The stirring speed is controlled at 50-100 r / min, and an intermittent stirring method is adopted. Stirring is performed for 30 minutes, stirring is stopped and the mixture is allowed to stand for 6 hours, and then stirred again for 30 minutes.

3. The refining process for hydrolyzed sponge according to claim 1, characterized in that: In step S30, the filter mesh size is preferably 100-300 mesh, the rinsing water is deionized water with a resistivity of not less than 15 MΩ·cm, and the standard for rinsing to neutral is that the pH value of the rinsing solution reaches 6.5-7.

5.

4. The refining process for hydrolyzed sponge according to claim 1, characterized in that: In step S40, the alkaline solution is selected from sodium hydroxide or potassium hydroxide solution, with a mass fraction preferably of 1%-1.5%, the neutralization temperature is controlled at 25-35℃, and the stirring speed is controlled at 30-50 r / min.

5. The refining process for hydrolyzed sponge according to claim 1, characterized in that: In step S60, the drying temperature is preferably 72-74℃, the drying time is 4-6 hours, and the drying equipment is a vacuum drying oven or a hot air circulating drying oven. The standard for drying to constant weight is that the difference between two consecutive weighings does not exceed 0.5%.

6. A method for applying a refining process to hydrolyzed sponges, used to achieve the refining process for hydrolyzed sponges as described in any one of claims 1-5, characterized in that, Includes the following steps: A10: Mix hydrolyzed sponge bone needles with a special solvent at a mass ratio of 1:1 to 1:3 to prepare a uniform paste. A20: Apply the paste to the skin surface and treat it using a specific transdermal massage technique for 3-5 minutes; A30: After removing surface powder, use the active ingredient preparation for subsequent care; A40: Establish a system for verifying the efficacy of sponge bone needles, including in vitro permeability testing and in vivo efficacy assessment; The specially prepared solvent contains glycerol, propylene glycol and deionized water in a mass ratio of 1:1:2 to 1:2:

3. The massage technique uses a reverse pore spiral lifting method, with pressure controlled within the range of 20-50 grams. The active ingredient formulation contains a functional ingredient with a molecular weight of 500-5000 Da and a mass fraction of 0.5%-5%.

7. The application method of the refining process for hydrolyzed sponge according to claim 6, characterized in that: In step A10, the special solvent also contains 0.1%-0.5% of a preservative and 0.01%-0.05% of an antioxidant. The preservative is selected from either phenoxyethanol or potassium sorbate, and the antioxidant is selected from either vitamin E or tert-butylhydroquinone.

8. The application method of the refining process for hydrolyzed sponge according to claim 6, characterized in that, In step A20, the massage techniques specifically include: First, apply 20-30 grams of pressure and massage in a circular motion for 1-2 minutes. Then, apply 30-50 grams of pressure and massage against the pores for 2-3 minutes. The massage frequency is 60-80 times per minute.

9. The application method of the refining process for hydrolyzed sponge according to claim 6, characterized in that, In step A40, the in vitro permeability test uses the Franz diffusion cell method: Using pig ear skin or artificial skin as a permeability barrier, the test temperature is 32-37℃, and the sampling time points are 15 minutes, 30 minutes, 1 hour, 2 hours, 4 hours and 6 hours. The detection method is HPLC or fluorescence spectrophotometry.

10. The application method of the refining process for hydrolyzed sponge according to claim 6, characterized in that, In step A40, the in vivo efficacy assessment includes: Animal models or human volunteers were used, and the evaluation indicators included skin hydration, transepidermal water loss, skin elasticity, melanin index, and hemoglobin index. The testing period was 1-4 weeks, and the data analysis used statistical methods with a p-value of less than 0.05.

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