Donkey-hide gelatin capable of promoting human cell repair function and preparation method of donkey-hide gelatin

Abstract

The invention relates to a preparation method of colla corii asini capable of promoting the repair function of human cells, which specifically comprises the following steps: in the colla corii asini preparation method, the preparation process is optimized, and the extraction is carried out twice in the extraction and filtration steps; repeating the steps of continuous concentration and foam extraction for eight times, adding filtered soybean oil after the sixth foam extraction, adding dissolved and filtered rock sugar after the seventh foam extraction, adding filtered yellow wine after the eighth foam extraction, and carrying out the ninth extraction until yellow fine foam appears on the surface of the glue solution, transferring the glue solution into a glue discharging pot, and preparing to obtain the donkey-hide gelatin capable of promoting the repairing function of human cells. Furthermore, according to application of the donkey-hide gelatin prepared according to the embodiment of the invention and a contrast in repairing human fibroblast injury caused by ultraviolet irradiation such as UVC, the donkey-hide gelatin prepared by the preparation method disclosed by the invention has a better effect in repairing human fibroblast injury caused by UVC, and has a great application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of collagen preparation, and in particular to donkey-hide gelatin that promotes the repair function of human cells and a preparation method thereof. Background Art

[0002] Donkey-hide gelatin is a traditional precious Chinese medicine. It is a solid gelatin made by decocting and concentrating the dried or fresh skin of the donkey Equus asinus L. of the equid family. It belongs to the lung, liver, and kidney meridians, and has the effects of nourishing blood and yin, moistening dryness, and stopping bleeding. It is used to treat blood deficiency, yin deficiency, bleeding and other related diseases. Modern pharmacological studies have shown that donkey-hide gelatin mainly contains protein and peptide components, and after hydrolysis, it obtains a variety of amino acids, as well as various trace elements that are beneficial to the human body. It has the effects of promoting hematopoiesis, reducing blood viscosity, resisting lung injury, enhancing immunity, anti-inflammatory, anti-tumor, and anti-shock. However, there are large differences in the efficacy of donkey-hide gelatin prepared by different methods, and the prior art still lacks reports on the repair of damage caused by ultraviolet rays, especially UVC, to human dermal cells. Summary of the invention

[0003] In view of the technical problems in the prior art such as lack of donkey-hide gelatin that can promote the repair function of human cells and a preparation method thereof, a donkey-hide gelatin that can promote the repair function of human cells and a preparation method thereof are provided.

[0004] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0005] The present invention provides a method for preparing donkey-hide gelatin that promotes the repair function of human cells, the method comprising the following steps:

[0006] Extraction and filtration: the process includes putting the blanched donkey skin into a container, adding water, heating and extracting twice, filtering and obtaining the glue juice;

[0007] Concentration, foaming and gelling: including initial concentration, continued concentration, foaming and gelling;

[0008] Initial concentration: The rubber juice is pumped into the concentrator for preliminary concentration. When the relative density of the rubber juice reaches 1.08-1.09, foaming begins. The floating black rubber foam is skimmed off for the first foaming, and then pumped into the tilting sandwich pot for continued concentration.

[0009] Continue to concentrate and remove foam: The gum juice continues to concentrate in the tilting jacketed pot, and the foam and impurities floating on the surface of the gum juice are skimmed off. This is repeated every 2 hours for eight times.

[0010] Glue production: Continue to concentrate and boil until it becomes a thick paste. When the specified glue water content is reached, transfer the glue into a condensation box pre-coated with vegetable oil.

[0011] Preferably, in the extraction and filtration steps, the first extraction is: adding water to the steaming ball at a mass ratio of wet skin to water of 1:0.6, measuring the water intake with a water meter, and closing the water inlet valve after reaching the specified water volume; then starting the steaming ball, passing clean steam, and controlling the pressure indicated by the pressure gauge at 0.12-0.15Mpa, first dynamically extracting for 1 hour, stopping the ball to vent once after 30 minutes, and then performing static extraction for 6 hours, rotating the steaming ball and stirring for 2-5 minutes every half an hour, and filtering the gum juice into a storage tank after the extraction is completed.

[0012] Preferably, in the extraction and filtration steps, the second extraction is as follows: water is added to the steaming ball at a mass ratio of wet skin to water of 1:0.5, the water inlet amount is measured with a water meter, and the water inlet valve is closed after the specified water amount is reached; the steaming ball is started, clean steam is introduced, the pressure indicated by the pressure gauge is controlled at 0.12-0.15Mpa, dynamic extraction is performed for 1 hour, the extracted gum juice is filtered, and combined with the gum juice filtrate of the first extraction, and the residue is removed after the extraction is completed. The extracted gum juice is filtered to obtain the gum juice after standing and clarifying.

[0013] Preferably, in the continuing concentration and frothing steps: in the continuing concentration and frothing steps: respectively add filtered soybean oil after the sixth frothing, add melted and filtered rock sugar after the seventh frothing, add filtered rice wine after the eighth frothing, and continue until yellow fine foam appears on the surface of the glue liquid after the ninth frothing, and then transfer the glue liquid to the glue discharge pot.

[0014] Preferably, in the glue discharging step, the prescribed glue discharging water content is 26wt%-29wt%.

[0015] Preferably, before the extraction and filtering steps, the following steps are also included:

[0016] Picking and weighing: Pick out the impurities in the donkey skin and weigh the donkey skin without impurities;

[0017] Soaking: Put the donkey skin into the soaking pool, soak it with drinking water for 4-8 days, change the water 1-3 times a day, and take it out of the pool after soaking until the skin layers are clearly stratified and white in color;

[0018] Shaving and skinning: the soaked donkey skin is shaved, then skinned and cut into pieces with a side length of 20-40 cm;

[0019] Washing leather: Put the leather pieces into the leather washing pool for washing;

[0020] Feeding and blanching: put the washed donkey hide into a steaming ball, add sodium carbonate at a ratio of 0.4%-0.65% of the wet hide weight, add water at a ratio of wet hide to water of 1:0.45-0.7, pass steam to heat and blanch the hide, control the steam pressure at 0.12-0.15Mpa, and blanch for 20-60min. Take out and rinse with water for standby use.

[0021] Preferably, the preparation method further comprises the following steps after the extraction and gelling steps:

[0022] Gel: Move the condensation box filled with glue liquid into the preliminary condensation workshop, and keep the glue liquid level in the condensation box horizontal, the temperature is 2-7℃, condense for 10-12h, and then move it into the second condensation workshop: the temperature is 0-2℃, condense to a certain hardness;

[0023] Cutting and tangent: Cut the film into pieces with a size of (8.8±0.1) cm×(3.6±0.1) cm. Cut the pieces in an environment with a temperature of 16-25°C and a relative humidity of 55-70% to divide the film into five equal parts.

[0024] Glue drying: Move the cut film to the glue drying room for drying. The drying temperature is 16-25℃, the relative humidity is 55%-70%, the cycle is 44-62 days, and the moisture content after drying is ≤15wt%;

[0025] Wipe the film: Wipe the film surface with a damp cloth to make it smooth and clean;

[0026] Sterilization: sterilize the film after rubbing with ultraviolet light; the sterilization temperature is controlled at 30-40℃, and the sterilization time is 20-40min;

[0027] Inner packaging: Pack the film according to the specification (31.25±9)g / piece, the packaging temperature is 18~26℃, and the relative humidity is 55%~65%;

[0028] Outer packaging: transfer the products that have been completed with inner packaging into boxes and pack them according to the specification of 8 pieces / box, then print and seal the boxes;

[0029] Inspection and warehousing: Inspection, and the finished products are put into storage after passing the inspection.

[0030] The present invention also provides donkey-hide gelatin obtained by any of the above-mentioned methods for preparing donkey-hide gelatin that promotes the repair function of human cells.

[0031] The present invention also provides the use of the donkey-hide gelatin in preparing a human dermal fibroblast damage repair product.

[0032] Preferably, the human dermal fibroblast damage is damage caused by ultraviolet radiation.

[0033] Preferably, the ultraviolet rays include any band of UVA, UVB, UVC and / or EUV.

[0034] Preferably, the product includes any one of food, health care product and medicine.

[0035] Compared with the prior art, the present invention has achieved the following beneficial technical effects: in the process of preparing the method for donkey-hide gelatin that promotes the repair function of human cells, by optimizing the preparation process, specifically including in the extraction and filtration steps, two extractions are performed; in the continuous concentration and foaming steps, eight times are repeated, and the filtered soybean oil is added after the sixth foaming, the crystal sugar after the melting and filtration is added after the seventh foaming, and the filtered yellow wine is added after the eighth foaming, until the ninth foaming until the surface of the glue liquid appears yellow fine foam, and then the glue liquid is transferred to the glue pot to prepare the donkey-hide gelatin that promotes the repair function of human cells. Further, the donkey-hide gelatin prepared by the embodiments of the present invention and the comparative examples is applied to the repair of human fibroblast damage caused by ultraviolet radiation such as UVC. It is found that the donkey-hide gelatin obtained by the preparation method of the present invention has a better effect in repairing human fibroblast damage caused by UVC, and has great application prospects. DETAILED DESCRIPTION

[0036] The specific embodiments of the present invention are described in detail below. It should be understood that the specific embodiments described herein are only used to illustrate and explain the present invention, and are not used to limit the present invention.

[0037] Unless otherwise specified, the reagents and experimental methods involved in the examples are conventional reagents and methods.

[0038] Human dermal fibroblasts (CP-H103, Wuhan Pronocell Life Science Co., Ltd.);

[0039] DMEM basal medium, trypsin (GI BCO), FBS, HEPES (HYCLONE), 2-mercaptoethanol, DMSO, LG ln, sodium pyruvate, penicillin / streptomycin (SI GMA), CCK-8 kit (Biyuntian).

[0040] Cell culture medium: DMEM basal medium, FBS 10%, HEPES 20mmo l / L, LG ln 0.2mmo l / L, sodium pyruvate 0.1mmo l / L, 2-mercaptoethanol 50nmo l / L, penicillin and streptomycin 100u / ml each, pH 7.2, 0.22μm filtration.

[0041] Example 1 A method for preparing donkey-hide gelatin that promotes the repair function of human cells

[0042] (1) Sorting and weighing: Pick out the impurities in the donkey hide and weigh 10 kg of donkey hide without impurities.

[0043] (2) Soaking: Put the donkey hide into a soaking pool and soak it with drinking water for 5 days. Change the water twice a day. Soak the donkey hide until the skin layers are clearly stratified and white in color, then remove it from the pool.

[0044] (3) Shaving and skinning: After the donkey skin is shaved, the skin is cut into pieces with a side length of about 30 cm.

[0045] (4) Washing leather: Put the leather pieces into the leather washing pool for washing.

[0046] (5) Adding materials and blanching: Put the cleaned donkey hide into a clean steaming ball, add sodium carbonate at a rate of 0.5% of the weight of the wet hide, add water at a mass ratio of wet hide to water of 1:0.5, pass steam to heat and blanch the hide, control the steam pressure at 0.12 MPa, and blanch for 40 minutes. The donkey hide will roll up, take it out and rinse it with water until the washing water passes the test and is ready for use.

[0047] (6) Extraction and filtration: Put the blanched donkey hide into the extraction equipment container, add water and heat to extract twice, filter to obtain gum juice, the first extraction: add water to the steaming ball according to the mass ratio of wet hide to water of 1:0.6, use a water meter to measure the water intake, and close the water inlet valve after reaching the specified water volume; start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.12Mpa. First, perform dynamic extraction for 1 hour, stop the ball to vent once every 30 minutes, and then perform static extraction. The static extraction lasts for 6 hours, and the steaming ball is rotated and stirred for 2 minutes every half an hour. After the extraction is completed, filter the gum juice into a storage tank.

[0048] Second extraction: add water to the steaming ball at a mass ratio of 1:0.5 between wet skin and water, measure the water intake with a water meter, close the water inlet valve after reaching the specified water volume, start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.12Mpa. Dynamic extraction for 1 hour, filter the extracted gum juice, and combine it with the gum juice filtrate from the first extraction. After the extraction is completed, remove the residue. After the extracted gum juice is allowed to stand for clarification, filter it to obtain the gum juice.

[0049] (7) Concentrated foaming and gelling:

[0050] Initial concentration: The rubber juice is pumped into the concentrator for preliminary concentration. When the relative density of the rubber juice reaches 1.08-1.09, start to froth. Use a stainless steel ladle to skim off the floating black rubber foam for the first froth extraction and pump it into a tilting sandwich pot for continued concentration.

[0051] Continue to concentrate and remove foam: The gum juice continues to concentrate in the tilting double-layer pot, and the foam and impurities floating on the surface of the gum juice are removed. The foam is removed every 2 hours and repeated eight times. After the sixth foaming, 320g of filtered soybean oil is added, after the seventh foaming, 320g of melted and filtered rock sugar is added, after the eighth foaming, 80g of filtered rice wine is added, and the mixture is stirred until fine yellow foam appears on the surface of the gum liquid after the ninth foaming. Then the gum liquid is transferred to the gum discharging pot.

[0052] Glue production: Continue to concentrate and boil until it becomes a thick paste, that is, when the specified glue water content reaches 26wt%, transfer the glue liquid into a stainless steel condensation box pre-coated with vegetable oil.

[0053] (8) Gel: Move the condensation box containing the gel liquid into the primary condensation workshop and keep the gel liquid level in the condensation box horizontal at a temperature of 2°C for condensation for 10 hours. Then move it into the secondary condensation workshop at a temperature of 0°C for condensation.

[0054] (9) Cutting and tangent: Cut the film into pieces with the size of (8.8±0.1) cm×(3.6±0.1) cm. Cut the pieces at a temperature of 16°C and a relative humidity of 55% and divide the film into five equal parts, making four cuts that can be broken by hand.

[0055] (10) Glue drying: Move the cut film to a glue drying room at a temperature of 16°C and a relative humidity of 55%. The cycle is 44 days and the moisture content is about 10.27 wt%.

[0056] (11) Wiping the film: Use a cloth moistened with purified water at about 60°C to wipe the film surface to make it smooth and clean, without yellow foam, fingerprints and white spots.

[0057] (12) Sterilization: The erased film is sterilized by ultraviolet irradiation; the temperature is controlled at 30°C during sterilization, and the sterilization time is 30 minutes.

[0058] (13) Inner packaging: Qualified films shall be packaged in PVC plates and aluminum foil with a specification of 31.25±9g / piece. Environmental humidity requirements: temperature is 18°C ​​and relative humidity is 55%.

[0059] (14) Outer packaging: Transfer the product into boxes according to the batch packaging instructions, 8 pieces / box, print the specified information on the outside of the box, and seal the box.

[0060] (15) Inspection and warehousing: Inspection is carried out according to enterprise standards and requirements, and qualified finished products are put into storage. Example 2 A method for preparing donkey-hide gelatin that promotes the repair function of human cells

[0061] (1) Sorting and weighing: Pick out the impurities in the donkey hide and weigh 10 kg of donkey hide without impurities.

[0062] (2) Soaking: Put the donkey hide into a soaking pool and soak it with drinking water for 5 days. Change the water twice a day. Soak the donkey hide until the skin layers are clearly stratified and white in color, then remove it from the pool.

[0063] (3) Shaving and skinning: After the donkey skin is shaved, the skin is cut into pieces with a side length of about 30 cm.

[0064] (4) Washing the leather: Put the leather into the leather washing pool and wash until it is clean.

[0065] (5) Adding materials and blanching: Put the cleaned donkey hide into a clean steaming ball, add sodium carbonate at a rate of 0.5% of the weight of the wet hide, add water at a ratio of 1:0.5, pass steam to heat and blanch the hide, control the steam pressure at 0.13 MPa, and blanch for 40 minutes. The donkey hide will roll up, take it out and rinse it with water until the washing water passes the test.

[0066] (6) Extraction and filtration: Put the blanched donkey skin into the extraction equipment container, add water and heat to extract twice, filter, and obtain the glue juice. For the first extraction: add water to the steaming ball at a ratio of 1:0.6 between wet skin and water, and use a water meter to measure the water intake. After reaching the specified water volume, close the water inlet valve. Start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.13 MPa. First, perform dynamic extraction for 1 hour, stop the ball to vent once after 30 minutes, and then perform static extraction for 6 hours. Rotate the steaming ball and stir for 3 minutes every half an hour. After the extraction is completed, filter the glue juice into a storage tank.

[0067] Second extraction: add water to the steaming ball at a ratio of 1:0.5 to wet skin, measure the water intake with a water meter, and close the water inlet valve after reaching the specified water volume. Start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.13Mpa. Dynamic extraction for 1 hour, filter the extracted gum juice, and combine it with the gum juice filtrate from the first extraction. After the extraction is completed, remove the residue. After the extracted gum juice is allowed to stand and clarify, filter it to obtain the gum juice.

[0068] (7) Concentrated foaming and gelling:

[0069] Initial concentration: The rubber juice is pumped into the concentrator for preliminary concentration. When the relative density of the rubber juice reaches 1.09, start to froth. Use a stainless steel ladle to skim off the floating black rubber foam for the first froth extraction and pump it into a tilting sandwich pot for continued concentration.

[0070] Continue to concentrate and remove foam: The gum juice continues to concentrate in the tilting double-layer pot, and the foam and impurities floating on the surface of the gum juice are removed. The foam is removed every 2 hours and repeated eight times. After the sixth foaming, 320g of filtered soybean oil is added, after the seventh foaming, 320g of melted and filtered rock sugar is added, after the eighth foaming, 80g of filtered rice wine is added, and the mixture is stirred until fine yellow foam appears on the surface of the gum liquid after the ninth foaming. Then the gum liquid is transferred to the gum discharging pot.

[0071] Glue output: Continue to concentrate and boil until it becomes a thick paste, that is, when the specified glue water content reaches 27wt%, transfer the glue liquid into a special stainless steel condensation box pre-coated with vegetable oil.

[0072] (8) Gel: Move the condensation box containing the glue liquid into the preliminary condensation workshop and keep the glue liquid level in the condensation box horizontal. The temperature is 5°C and condensation is carried out for 11 hours. Then move it into the second condensation workshop: the temperature is 1°C and condense to the appropriate hardness.

[0073] (9) Cutting and cutting: Cut the film into pieces, the film size is: (8.8±0.1) cm×(3.6±0.1) cm, the temperature is 20°C, the relative humidity is 60%, and then cut the line: divide the film into 5 equal parts, make 4 cuts, and break it by hand.

[0074] (10) Glue drying: Move the cut film to the glue drying room at a temperature of 20°C and a relative humidity of 60%. The production cycle is 55 days and the moisture content is about 10.64 wt%.

[0075] (11) Wiping: Wipe the film surface with a cloth moistened with purified water at about 60°C to make the film surface smooth and clean, without yellow foam, fingerprints, or white spots, and with traditional cloth texture.

[0076] (12) Sterilization: The erased film is sterilized by ultraviolet irradiation; the temperature is controlled at 35°C during sterilization, and the sterilization time is 30 minutes.

[0077] (13) Inner packaging: Qualified films shall be packaged in PVC plates and aluminum foil, 31.25±9g / piece. Environmental humidity requirements: temperature is 22°C and relative humidity is 60%.

[0078] (14) Outer packaging: Transfer the product into boxes according to the batch packaging instructions, 8 pieces / box, print the specified information on the outside of the box, and seal the box.

[0079] (15) Inspection and warehousing: Inspection is carried out according to enterprise standards and requirements, and qualified finished products are put into storage. Example 3 A method for preparing donkey-hide gelatin that promotes the repair function of human cells

[0080] (1) Sorting and weighing: Pick out the impurities in the donkey hide and weigh 10 kg of donkey hide without impurities.

[0081] (2) Soaking: Put the donkey hide into a soaking pool and soak it with drinking water for 5 days. Change the water twice a day. Soak the donkey hide until the skin layers are clearly stratified and white in color, then remove it from the pool.

[0082] (3) Shaving and skinning: After the donkey skin is shaved, the skin is cut into pieces with a side length of about 30 cm.

[0083] (4) Washing the leather: Put the leather into the leather washing pool and wash until it is clean.

[0084] (5) Adding materials and blanching: Put the cleaned donkey hide into a clean steaming ball, add sodium carbonate at a rate of 0.5% of the weight of the wet hide, add water at a ratio of 1:0.5, pass steam to heat and blanch the hide, control the steam pressure at 0.15 MPa, and blanch for 40 minutes. The donkey hide will roll up, take it out and rinse it with water until the washing water passes the test.

[0085] (6) Extraction and filtration: Put the blanched donkey skin into the extraction equipment container, add water and heat to extract twice, filter, and obtain the glue juice. For the first extraction: add water to the steaming ball at a ratio of 1:0.6 between wet skin and water, and use a water meter to measure the water intake. After reaching the specified water volume, close the water inlet valve. Start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.15 MPa. First, perform dynamic extraction for 1 hour, stop the ball to vent once after 30 minutes, and then perform static extraction. The static extraction lasts for 6 hours, and the steaming ball is rotated and stirred for 5 minutes every half an hour. After the extraction is completed, filter the glue juice into a storage tank.

[0086] Second extraction: add water to the steaming ball at a ratio of 1:0.5 to wet skin, use a water meter to measure the water intake, close the water inlet valve after reaching the specified water volume, start the steaming ball, pass clean steam, and control the pressure indicated by the pressure gauge at 0.15Mpa. Dynamic extraction for 1 hour, filter the extracted gum juice, and combine it with the gum juice filtrate from the first extraction. After the extraction is completed, remove the residue. After the extracted gum juice is allowed to stand and clarify, filter it to obtain the gum juice.

[0087] (7) Concentrated foaming and gelling:

[0088] Initial concentration: The rubber juice is pumped into the concentrator for preliminary concentration. When the relative density of the rubber juice reaches 1.09, start to froth. Use a stainless steel ladle to skim off the floating black rubber foam for the first froth extraction and pump it into a tilting sandwich pot for continued concentration.

[0089] Continue to concentrate and remove foam: The gum juice continues to concentrate in the tilting double-layer pot, and the foam and impurities floating on the surface of the gum juice are removed. The foam is removed every 2 hours and repeated eight times. After the sixth foaming, 320g of filtered soybean oil is added, after the seventh foaming, 320g of melted and filtered rock sugar is added, after the eighth foaming, 80g of filtered rice wine is added, and the mixture is stirred until fine yellow foam appears on the surface of the gum liquid after the ninth foaming. Then the gum liquid is transferred to the gum discharging pot.

[0090] Glue production: Continue to concentrate and boil until it becomes a thick paste. That is, when the specified glue water content reaches 29wt%, transfer the glue liquid into a special stainless steel condensation box pre-coated with vegetable oil.

[0091] (8) Gel: Move the condensation box containing the glue liquid into the preliminary condensation workshop and keep the glue liquid level in the condensation box horizontal. The temperature is 7°C and condensation is carried out for 12 hours. Then move it into the second condensation workshop: the temperature is 2°C and condense to the appropriate hardness.

[0092] (9) Cutting and cutting: Cut the film into pieces, the film size is: (8.8±0.1) cm×(3.6±0.1) cm, the temperature is 25°C, the relative humidity is 70%, and then cut the film: divide the film into 5 equal parts, make 4 cuts, and break it by hand.

[0093] (10) Glue drying: Move the cut film to the glue drying room at a temperature of 25°C and a relative humidity of 70%. The production cycle is 62 days and the moisture content is about 11.05 wt%.

[0094] (11) Wiping: Wipe the film surface with a cloth moistened with purified water at about 60°C to make the film surface smooth and clean, without yellow foam, fingerprints, or white spots, and with traditional cloth texture.

[0095] (12) Sterilization: The film after erasing is sterilized by ultraviolet irradiation; the temperature during sterilization is controlled at 40°C and the sterilization time is 30 minutes.

[0096] (13) Inner packaging: Qualified films shall be packaged in PVC plates and aluminum foil, 31.25±9g / piece. Environmental humidity requirements: temperature is 26°C and relative humidity is 65%.

[0097] (14) Outer packaging: Transfer the product into boxes according to the batch packaging instructions, 8 pieces / box, print the specified information on the outside of the box, and seal the box.

[0098] (15) Inspection and warehousing: Inspection is carried out in accordance with enterprise standards and requirements, and qualified finished products are put into storage.

[0099] The difference between Comparative Example 1 and Example 1 is that in the (6) extraction and filtration step, water is added and heated for extraction once, that is, only the first extraction is performed, and the rest is the same as Example 1.

[0100] The difference between Comparative Example 2 and Example 1 is that in the step (7) of concentrating, extracting foam and discharging glue, the foam is extracted once every hour, and the rest is the same as Example 1.

[0101] The difference between Comparative Example 3 and Example 1 is that in the step (7) of concentration, frothing and gelling, the process of concentrating and frothing is repeated three times, and 320 g of filtered soybean oil, 320 g of filtered crystal sugar and 80 g of filtered rice wine are added at one time after the last frothing. The rest is the same as Example 1.

[0102] Experimental Example 1: Test on the repair of UVC damage to cells by donkey-hide gelatin

[0103] 2 g, 4 g, 8 g, 10 g, and 15 g of donkey-hide gelatin powder (the donkey-hide gelatin block of Example 1 was made into powder) were weighed respectively and added into 100 mL of culture medium to obtain a culture medium containing donkey-hide gelatin (0.02 g / mL, 0.04 g / mL, 0.08 g / mL, 0.1 g / mL, and 0.15 g / mL).

[0104] The frozen human dermal fibroblasts were revived and cultured, and the well-growing cells were cultured at 1.0×10 4cels / well were plated in a 96-well culture plate and cultured at 37°C, 5% CO2 for 24 hours. The supernatant was aspirated, the cells were washed twice with PBS, and 0.1 mL of culture medium containing donkey-hide gelatin (0 g / mL (model group), 0.02 g / mL, 0.04 g / mL, 0.08 g / mL, 0.10 g / mL, and 0.15 g / mL) of Example 1 was added to each well. The cells were then exposed to a UVC semi-lethal dose (5 J / cm 2 ) for 75 min, and culture wells without irradiation and without donkey-hide gelatin were set as the control group. After irradiation, the OD value was determined by CCK-8 method for 24 h, and the cell survival rate was calculated. Each group was repeated three times and the average value was taken. The test results are shown in Table 1.

[0105] Table 1 Repair effect of different concentrations of donkey-hide gelatin on UVC-damaged human dermal fibroblasts

[0106]

[0107]

[0108] Note: Compared with the control group: ## p<0.01; compared with the model group: * p<0.05, ** p<0.01.

[0109] The results in Table 1 show that compared with the control group, the survival rate of human dermal fibroblasts in the model group was significantly reduced, indicating that the UVC damage model was successfully established; compared with the model group, the culture medium containing different concentrations of donkey-hide gelatin can significantly repair the damage caused by UVC to human dermal fibroblasts, among which the repair effect of 0.08g / mL donkey-hide gelatin was the most significant.

[0110] Experimental Example 2: Test II on the repair of UVC damage to cells by donkey-hide gelatin

[0111] The frozen human dermal fibroblasts were revived and cultured, and the well-growing cells were cultured at 1.0×10 4 cels / well were seeded in a 96-well culture plate and cultured at 37°C, 5% CO2 for 24 hours, the supernatant was aspirated, the cells were washed twice with PBS, 0.1 mL of culture medium containing 0 g / mL (model group) and 0.08 g / mL donkey-hide gelatin (Example 1-3, Comparative Example 1-3) was added to each well, the cells were irradiated with a semi-lethal dose of UV C (5 J / cm2) for 75 min, and culture wells without irradiation and without donkey-hide gelatin were set as a control group. After irradiation, the cells were cultured for 24 hours and the OD value was determined by CCK-8 method to calculate the cell survival rate. Each group was repeated three times and the average value was taken. The test results are shown in Table 2.

[0112] Table 2 Repair effect of different groups of donkey-hide gelatin culture on UVC-damaged human dermal fibroblasts

[0113]

[0114]

[0115] Note: Compared with the control group: ## p<0.01; compared with the model group: * p<0.05, ** p<0.01; compared with the control group: a p<0.05.

[0116] The results in Table 2 show that compared with the control group, the survival rate of human dermal fibroblasts in the model group was significantly reduced, indicating that the UVC damage model was successfully established; compared with the model group, the culture medium containing donkey-hide gelatin can significantly repair the damage caused by UVC to human dermal fibroblasts, among which the repair effect of donkey-hide gelatin in Example 1-3 group is the most significant; compared with Comparative Examples 1-3, the culture medium containing donkey-hide gelatin in Examples 1-3 can better repair the damage caused by UVC to human dermal fibroblasts.

[0117] The present invention illustrates the technical solution of the present invention through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned steps, that is, it does not mean that the present invention must rely on the above-mentioned steps to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials selected by the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

Claims

1. A method for preparing donkey-hide gelatin that promotes the repair function of human cells, characterized in that The method comprises the following steps: Extraction and filtration: the process includes putting the blanched donkey skin into a container, adding water, heating and extracting twice, filtering and obtaining the glue juice; Concentration, foaming and gelling: including initial concentration, continued concentration, foaming and gelling; Initial concentration: The rubber juice is pumped into the concentrator for preliminary concentration. When the relative density of the rubber juice reaches 1.08-1.09, foaming begins. The floating black rubber foam is skimmed off for the first foaming, and then pumped into the tilting sandwich pot for continued concentration. Continue to concentrate and remove foam: The gum juice continues to concentrate in the tilting jacketed pot, and the foam and impurities floating on the surface of the gum juice are skimmed off. This is repeated every 2 hours for eight times. Glue production: Continue to concentrate and boil until it becomes a thick paste. When the specified glue water content is reached, transfer the glue liquid into a condensation box pre-coated with vegetable oil.

2. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, wherein in the extraction and filtration steps, the first extraction is: adding water to the steaming ball at a mass ratio of wet skin to water of 1:0.6, measuring the water intake with a water meter, and closing the water inlet valve after reaching the specified water volume; then starting the steaming ball, passing clean steam, and controlling the pressure indicated by the pressure gauge at 0.12-0.15Mpa, first dynamically extracting for 1 hour, stopping the ball to vent once after 30 minutes, and then performing static extraction, which takes 6 hours, rotating the steaming ball and stirring for 2-5 minutes every half an hour, and filtering the gelatin into a storage tank after the extraction is completed.

3. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, wherein in the extraction and filtration steps, the second extraction is as follows: water is added to the steam bulb at a mass ratio of wet skin to water of 1:0.5, the water intake is measured with a water meter, and the water inlet valve is closed after the specified water volume is reached; the steam bulb is started, clean steam is introduced, and the pressure indicated by the pressure gauge is controlled at 0.12-0.15Mpa, and dynamic extraction is performed for 1 hour, the extracted gelatin is filtered, and the extracted gelatin is combined with the gelatin filtrate of the first extraction, and the residue is removed after the extraction is completed. The extracted gelatin is filtered to obtain the gelatin after standing and clarifying.

4. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, wherein in the steps of continuous concentration and foaming: filtered soybean oil is added after the sixth foaming, melted and filtered rock sugar is added after the seventh foaming, and filtered rice wine is added after the eighth foaming, and the mixture is stirred until yellow fine foam appears on the surface of the gelatin liquid after the ninth foaming, and then the gelatin liquid is transferred to a gelatin discharging pot.

5. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, wherein in the gelatin discharging step, the prescribed gelatin discharging water content is 26wt%-29wt%.

6. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, before the extraction and filtration steps, further comprising the following steps: Picking and weighing: Pick out the impurities in the donkey skin and weigh the donkey skin without impurities; Soaking: Put the donkey skin into the soaking pool, soak it with drinking water for 4-8 days, change the water 1-3 times a day, and take it out of the pool after soaking until the skin layers are clearly stratified and white in color; Shaving and skinning: the soaked donkey skin is shaved, then skinned and cut into pieces with a side length of 20-40 cm; Washing leather: Put the leather pieces into the leather washing pool for washing; Feeding and blanching: put the washed donkey hide into a steaming ball, add sodium carbonate at a ratio of 0.4%-0.65% of the wet hide weight, add water at a ratio of wet hide to water of 1:0.45-0.7, pass steam to heat and blanch the hide, control the steam pressure at 0.12-0.15Mpa, and blanch for 20-60min. Take out and rinse with water for standby use.

7. The method for preparing donkey-hide gelatin that promotes the repair function of human cells according to claim 1, wherein after the extraction and gelatinization steps, the method further comprises the following steps: Gel: Move the condensation box filled with glue liquid into the preliminary condensation workshop, and keep the glue liquid level in the condensation box horizontal, the temperature is 2-7℃, condense for 10-12h, and then move it into the second condensation workshop: the temperature is 0-2℃, condense to a certain hardness; Cutting and tangent: Cut the film into pieces with a size of (8.8±0.1) cm×(3.6±0.1) cm. Cut the pieces in an environment with a temperature of 16-25°C and a relative humidity of 55-70% to divide the film into five equal parts. Glue drying: Move the cut film to the glue drying room for drying. The drying temperature is 16-25℃, the relative humidity is 55%-70%, the cycle is 44-62 days, and the moisture content after drying is ≤15wt%; Wipe the film: Wipe the film surface with a damp cloth to make it smooth and clean; Sterilization: sterilize the film after rubbing with ultraviolet light; the sterilization temperature is controlled at 30-40℃, and the sterilization time is 20-40min; Inner packaging: Pack the film according to the specification (31.25±9)g / piece, the packaging temperature is 18~26℃, and the relative humidity is 55%~65%; Outer packaging: transfer the products that have been completed with inner packaging into boxes and pack them according to the specification of 8 pieces / box, then print and seal the boxes; Inspection and warehousing: Inspection, and the finished products are put into storage after passing the inspection.

8. A donkey-hide gelatin obtained by the method for preparing donkey-hide gelatin that promotes the repair function of human cells as claimed in any one of claims 1 to 7.

9. The use of donkey-hide gelatin according to claim 8 in the preparation of a human dermal fibroblast damage repair product, characterized in that: The product includes any one of food, health care products and medicines.

10. The use according to claim 9, wherein the human dermal fibroblast damage is caused by ultraviolet radiation, and the ultraviolet rays include any band of UVA, UVB, UVC and / or EUV.

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