Deer tendon composition as well as preparation method and application thereof

Abstract

The invention mainly relates to a deer tendon composition as well as a preparation method and application thereof, and belongs to the technical field of biology. The composition is prepared from 50-60 parts of deer tendons, 15-25 parts of mushroom, 6-10 parts of fructus lycii, 10-20 parts of millet and 1-3 parts of sodium alginate. The deer tendon composition is prepared by soaking deer tendons in honey, freeze-drying and crushing, freeze-drying and crushing Taiwan mushroom and fructus lycii respectively, parching and crushing millet, and mixing the crushed raw materials with sodium alginate. The deer tendon composition can improve the moisture and protein content of skin, increase the content of hydroxyproline and hyaluronic acid in the skin, improve the collagen binding capacity of aged skin and effectively repair oxidative damage caused by aging; meanwhile, the elasticity and glossiness of the skin can be improved, the yellowness and melanin are reduced, facial wrinkles are reduced, and the effects of resisting aging, resisting oxidation, maintaining beauty and keeping young are achieved.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a deer tendon composition and a preparation method and application thereof. Background Art

[0002] Deer tendon, also known as deer tendon, is a natural food obtained from deer hooves. It is warm in nature and has the effects of strengthening tendons and bones, producing essence and nourishing marrow. It is mainly used for the treatment of liver and kidney deficiency, strain and injury, wind-cold arthralgia, cramps, back pain, muscle fatigue or weakness, difficulty walking, weakness in hands and feet, and other diseases. At the same time, deer tendon is also rich in nutrients such as protein, collagen, and amino acids, and has application values ​​such as anti-oxidation and anti-aging, maintaining skin elasticity, moisturizing the skin, and repairing damaged tissues; due to the inconvenience of the source and use of deer tendon, it has not been widely used at present. The conventional deer tendon processing method is oil-making, water-making, and drying, which easily leads to the loss of nutrients, affects the nutritional value, and thus reduces its anti-aging, anti-oxidation, and beauty-enhancing effects. And its use alone is often poor in anti-aging, anti-oxidation, and beauty-enhancing effects due to its single composition. Therefore, it is particularly important to provide a composition and preparation method for anti-aging, anti-oxidation, and beauty-enhancing that can improve the efficacy of deer tendon.

[0003] Tai Mushroom is produced in Mount Wutai and is one of the advanced edible fungi. It contains 18 kinds of amino acids and multiple vitamins needed by the human body. It has the effects of lowering cholesterol, preventing kidney disease, gallstones, diabetes, cirrhosis, liver cancer, etc. It is an ideal food for strengthening physical fitness. Millet is a common food with the effects of strengthening the spleen and stomach, replenishing deficiency, harmonizing the middle and benefiting the kidney, removing heat and detoxifying. Lycium barbarum is a kind of raw material with medicinal and edible properties. It has the effects of nourishing the liver and kidney, benefiting the essence and improving the eyesight. It is often used for symptoms such as consumptive cough, dizziness, yin deficiency, liver and kidney deficiency, backache and knee weakness, thirst, and spermatorrhea. Sodium alginate is a natural polymer present in brown algae, which has the effects of anti-tumor, regulating immunity, resisting hyperlipidemia, lowering blood sugar, and resisting radiation. At present, there is no report on the content that the combination of Tai Mushroom, millet, wolfberry, sodium alginate and deer tendon can enhance the anti-aging, antioxidant and beauty effects of deer tendon. Summary of the invention

[0004] In view of this, the purpose of the present invention is to provide a deer tendon composition and a preparation method, which, on the basis of better retaining the functional components of each raw material, enables the synergistic effect between the raw materials of the formula to jointly achieve the purpose of increasing the content of skin moisture, protein, hydroxyproline, and hyaluronic acid, repairing oxidative damage caused by aging, improving skin elasticity and glossiness, reducing yellowness and melanin, reducing facial wrinkles, and effectively exerting anti-aging, anti-oxidation and beauty-enhancing effects.

[0005] The invention provides a deer tendon composition, which comprises the following raw materials in parts by weight: 50-60 parts of deer tendon, 15-25 parts of mushrooms, 6-10 parts of wolfberries, 10-20 parts of millet and 1-3 parts of sodium alginate.

[0006] Preferably, the deer tendon composition comprises the following raw materials in parts by weight: 53-57 parts of deer tendon, 18-22 parts of Agaricus oleifera, 7-9 parts of wolfberry, 13-17 parts of millet, and 1.5-2.5 parts of sodium alginate.

[0007] The present invention also provides a method for preparing the deer tendon composition, comprising the following steps:

[0008] Deer tendon is soaked in honey, freeze-dried, crushed and sieved to obtain deer tendon powder;

[0009] The mushrooms are freeze-dried, crushed and sieved to obtain mushroom powder;

[0010] The millet is roasted, crushed and sieved to obtain millet flour;

[0011] Lycium barbarum is freeze-dried, crushed and sieved to obtain Lycium barbarum powder;

[0012] Deer tendon powder, Taiwan mushroom powder, millet powder, wolfberry powder and sodium alginate are mixed to obtain a deer tendon composite.

[0013] Preferably, the honey soaking time is 1 to 3 hours.

[0014] Preferably, freeze-drying of deer tendon comprises the following steps: freezing the deer tendon soaked in honey at -40°C to -50°C for 2 to 4 hours; then vacuum freezing at -45°C to -55°C for 0.5 to 2.5 hours; and then heating until the pressure reaches 28 to 32 Pa and maintaining for 6 to 8 hours.

[0015] Preferably, freeze-drying of Agaricus oleraceus and wolfberry comprises the following steps: freezing Agaricus oleraceus or wolfberry at -40℃~-50℃ for 1~3h; then vacuum freezing at -45℃~-55℃ for 0.5~1.5h; then heating to the pressure reaching 28~32Pa and maintaining for 4~6h.

[0016] Preferably, the millet is roasted at 140-160° C. for 3-5 minutes.

[0017] Preferably, the sieving is that the deer tendons, Taiwanese mushrooms or wolfberries are sieved through a 40-70 mesh sieve, and the millet is sieved through a 40-50 mesh sieve.

[0018] Preferably, the mixing is to mix deer hoof tendon powder, Taiwan mushroom powder, millet powder, wolfberry powder and then mix with sodium alginate.

[0019] The present invention also provides application of the deer tendon composition or the preparation method in preparing anti-aging, anti-oxidation and / or beauty products.

[0020] Beneficial effects of the present invention:

[0021] The deer tendon composition of the present invention can adhere to and wrap other raw materials by adding millet to form a paste-like liquid with uniform concentration. At the same time, the addition of sodium alginate can also promote the formation of a viscous and uniform paste-like liquid, which is convenient for taking. Sodium alginate also has a film-forming effect. After meeting water, it can form a protective film on the surface of other raw material powder particles, effectively preventing the effective ingredients of other raw materials from being destroyed or lost during the process of taking it with water or simmering it. In addition, the freeze-drying of the present invention also better retains the effective ingredients of the raw materials. It makes the raw materials of the formula work better together to achieve the effects of anti-aging, anti-oxidation and beauty and skin care. It is specifically manifested in increasing the moisture and protein content of the skin, increasing the content of hydroxyproline and hyaluronic acid in the skin, improving the collagen binding ability of aging skin, and effectively repairing the oxidative damage caused by aging; improving the elasticity and glossiness of the skin, reducing yellowness and melanin, and reducing facial wrinkles. DETAILED DESCRIPTION

[0022] The invention provides a deer tendon composition, comprising the following raw materials in parts by weight: 50-60 parts of deer tendon, 15-25 parts of mushrooms, 6-10 parts of wolfberries, 10-20 parts of millet, and 1-3 parts of sodium alginate; preferably: 53-57 parts of deer tendon, 18-22 parts of mushrooms, 7-9 parts of wolfberries, 13-17 parts of millet, and 1.5-2.5 parts of sodium alginate; more preferably: 55 parts of deer tendon, 20 parts of mushrooms, 8 parts of wolfberries, 15 parts of millet, and 2 parts of sodium alginate.

[0023] The present invention has no special limitation on the sources of the deer tendons, mushrooms, wolfberries, millet and sodium alginate, and conventional commercial products in the art can be used.

[0024] The present invention also provides a method for preparing the deer tendon composition, comprising the following steps:

[0025] Deer tendon is soaked in honey and then freeze-dried, crushed and sieved to obtain deer tendon powder. The honey is preferably honey with a mass concentration of 10% to 20%. In one embodiment, honey with a concentration of 10%, 15% or 20% can be selected. The honey soaking time is preferably 1 to 3 hours, more preferably 1.5 hours to 2.5 hours, and further preferably 2 hours. The freeze-drying preferably includes the following steps: freezing the deer tendon soaked in honey at -40℃ to -50℃ for 2 to 4 hours; then vacuum freezing at -45℃ to -55℃ for 0.5 to 2.5 hours, and sublimation drying; then heating to a pressure of 28 to 32Pa, and maintaining for 6 to 8 hours, so as to effectively capture the sublimated vapor. In one embodiment, the freeze drying can be selected as follows: the deer tendon soaked in honey is frozen at -40°C, -41°C, -42°C, -43°C, -44°C, -45°C, -46°C, -47°C, -48°C, -49°C or -50°C for 2, 2.5, 3, 3.5 or 4 hours; then vacuum frozen at -45°C, -46°C, -47°C, -48°C, -49°C, -50°C, -51°C, -52°C, -53°C, -54°C or -55°C for 0.5h, 1h, 1.5h, 2h or 2.5h; then heated to a pressure of 28Pa, 29Pa, 30Pa, 31Pa or 32Pa, and maintained for 6h, 6.5h, 7h, 7.5h or 8h. After freeze drying, it is crushed. The present invention does not specifically limit the crushing method. In one embodiment, grinding and crushing can be selected. After crushing, the powder is sieved, preferably through a 40-70 mesh sieve. In one embodiment, a 40-mesh, 50-mesh, 60-mesh or 70-mesh sieve can be selected. The large particles that do not meet the particle size requirements obtained by sieving can be preferably repeated after the above crushing and sieving steps, and the obtained powders are mixed together to obtain deer tendon powder.

[0026] The mushrooms are freeze-dried, crushed and sieved to obtain mushroom powder. The freeze-drying preferably includes the following steps: freezing the mushrooms at -40℃ to -50℃ for 1 to 3 hours, then vacuum freezing at -45℃ to -55℃ for 0.5 to 1.5 hours, and sublimation drying; then heating to a pressure of 28 to 32 Pa, maintaining for 4 to 6 hours, so as to effectively capture the sublimated vapor. In one embodiment, the mushrooms can be frozen at -40°C, -41°C, -42°C, -43°C, -44°C, -45°C, -46°C, -47°C, -48°C, -49°C or -50°C for 1, 1.5, 2, 2.5 or 3 hours; then vacuum frozen at -45°C, -46°C, -47°C, -48°C, -49°C, -50°C, -51°C, -52°C, -53°C, -54°C or -55°C for 0.5h, 1h or 1.5h; then heated to a pressure of 28Pa, 29Pa, 30Pa, 31Pa or 32Pa, and maintained for 4h, 4.5h, 5h, 5.5h or 6h. After freeze drying, the mushrooms are crushed. The present invention does not specifically limit the crushing method. In one embodiment, grinding and crushing can be selected. After crushing, the powder is sieved, preferably through a 40-70 mesh sieve. In one embodiment, a 40-mesh, 50-mesh, 60-mesh or 70-mesh sieve can be selected. The large particles that do not meet the particle size requirements obtained by sieving can be preferably repeated after the above crushing and sieving steps, and the obtained powders are mixed together to obtain Taiwan mushroom powder.

[0027] The millet is roasted, crushed and sieved to obtain millet powder. The millet is preferably roasted at 140-160°C for 3-5 minutes to make the millet lightly burnt. In one embodiment, the millet can be roasted at 140°C, 145°C, 150°C, 155°C or 160°C for 3 minutes, 4 minutes or 5 minutes. After roasting, it is crushed. The present invention does not specifically limit the method of crushing. In one embodiment, grinding and crushing can be selected. After crushing, it is sieved. The sieving is preferably sieved through a 40-50 mesh sieve. In one embodiment, a 40 mesh, 45 mesh or 50 mesh sieve can be selected. The large particles that do not meet the particle size requirements obtained by sieving can be preferably repeated after the above-mentioned crushing and sieving steps, and the obtained powders are mixed together to obtain millet powder.

[0028] The wolfberry is freeze-dried, crushed and sieved to obtain wolfberry powder. The specific preparation method of the wolfberry powder is the same as the preparation method of the Taiwan mushroom powder.

[0029] Deer tendon powder, mushroom powder, millet powder, wolfberry powder and sodium alginate are mixed to obtain a deer tendon composition. The mixing is preferably mixing deer tendon powder, mushroom powder, millet powder and wolfberry powder and then mixing with sodium alginate.

[0030] The present invention also provides the use of the deer tendon composition or the preparation method in the preparation of anti-aging, anti-oxidation and / or beauty products. The deer tendon composition can increase the moisture and protein content of the skin, increase the content of hydroxyproline and hyaluronic acid in the skin, improve the collagen binding ability of aging skin, and effectively repair the oxidative damage caused by aging; at the same time, it can also improve the elasticity and glossiness of the skin, reduce yellowness and melanin, and reduce facial wrinkles, achieving the effects of anti-aging, anti-oxidation and beauty.

[0031] When the deer tendon composition is used, preferably a certain amount of the composition can be added to boiling water or water at room temperature and then simmered for 5 to 10 minutes, and then taken after cooling.

[0032] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0033] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0034] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0035] Example 1

[0036] Preparation of deer tendon composition

[0037] Formula: 55g deer tendon, 20g Taiwan mushroom, 8g wolfberry, 15g millet, and 2g sodium alginate.

[0038] Preparation method:

[0039] 1. Weigh the fresh deer tendon after purification, soak it in 15% honey for 2 hours, and drain. Freeze drying: adjust the temperature of the freeze drying equipment drying box plate layer and the product to -45°C, so that the liquid component in the deer tendon reaches the eutectic point, freeze drying for 3 hours; adjust the temperature to -50°C and turn on the vacuum pump, freeze drying for 1.5 hours; then slowly heat the plate layer until the pressure in the drying box reaches 30Pa, and maintain the effective pressure for 7 hours; after the moisture content is qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 50-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain deer tendon powder.

[0040] 2. Weigh the washed and dried mushrooms and freeze-dry them: adjust the temperature of the freeze-drying equipment drying box plate layer and the product to -45°C, so that the liquid component in the mushrooms reaches the eutectic point, and freeze-dry for 2 hours; then adjust the temperature to -50°C and turn on the vacuum pump, freeze-dry for 1 hour; then slowly heat the plate layer until the pressure in the drying box reaches 30Pa, and maintain the effective pressure for 5 hours; after the moisture content is qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 50-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain mushroom powder.

[0041] 3. Weigh millet, fry it in a drum frying pan at 150° C. for 3 min, crush it into powder, pass it through a 45-mesh sieve, and repeat the above crushing and sieving steps for the large particles that do not meet the particle size requirements. Then, mix the obtained powders together to obtain millet flour.

[0042] 4. Weigh the washed and dried wolfberries, freeze-dry them (the specific steps are the same as 2. of the present embodiment), grind them into powder after the freeze-dried moisture content is qualified (the mass fraction of moisture is ≤18%), pass through a 50-mesh sieve, and sieve the large particles that do not meet the particle size requirements. After repeating the above-mentioned crushing and sieving steps, the obtained powders are mixed together to obtain wolfberry powder.

[0043] 5. Mix the deer hoof tendon powder, Taiwanese mushroom powder, millet powder and wolfberry powder together and stir evenly.

[0044] 6. Add sodium alginate and stir evenly again to obtain a deer tendon composition.

[0045] Example 2

[0046] Preparation of deer tendon composition

[0047] Formula: 50g deer tendon, 25g Taiwan mushroom, 6g wolfberry, 20g millet, and 3g sodium alginate.

[0048] Preparation method:

[0049] 1. Weigh the purified fresh deer tendon, soak it in 10% honey for 1 hour, and drain. Freeze drying: adjust the temperature of the freeze drying equipment drying box plate layer and the product to -40°C, so that the liquid component in the deer tendon reaches the eutectic point, freeze drying for 4 hours; adjust the temperature to -45°C and turn on the vacuum pump, freeze drying for 2.5 hours; then slowly heat the plate layer until the pressure in the drying box reaches 28Pa, maintain the effective pressure for 8 hours; after the moisture content is qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 40-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain deer tendon powder.

[0050] 2. Weigh the washed and dried mushrooms and freeze-dry them: adjust the temperature of the freeze-drying equipment drying box plate layer and the product to -40°C, so that the liquid component in the mushrooms reaches the eutectic point, and freeze-dry for 3 hours; then adjust the temperature to -45°C and turn on the vacuum pump, freeze-dry for 1.5 hours; then slowly heat the plate layer until the pressure in the drying box reaches 28Pa, and maintain the effective pressure for 6 hours; after the moisture content is qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 40-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain mushroom powder.

[0051] 3. Weigh the millet, fry it in a drum frying pan at 140°C for 4 minutes, crush it into powder, pass it through a 40-mesh sieve, and repeat the above crushing and sieving steps for the large particles that do not meet the particle size requirements. Then, mix the obtained powders together to obtain millet flour.

[0052] 4. Weigh the washed and dried wolfberries, freeze-dry them (the specific steps are the same as 2. of the present embodiment), grind them into powder after the moisture content is qualified (the mass fraction of moisture is ≤18%) after freeze-drying, pass through a 40-mesh sieve, and sieve the large particles that do not meet the particle size requirements. After repeating the above-mentioned crushing and sieving steps, the obtained powders are mixed together to obtain wolfberry powder.

[0053] 5. Mix the deer hoof tendon powder, Taiwanese mushroom powder, millet powder and wolfberry powder together and stir evenly.

[0054] 6. Add sodium alginate and stir evenly again to obtain a deer tendon composition.

[0055] Example 3

[0056] Preparation of deer tendon composition

[0057] Formula: 60g deer tendon, 15g Taiwan mushroom, 10g wolfberry, 10g millet, and 1g sodium alginate.

[0058] Preparation method:

[0059] 1. Weigh the purified fresh deer tendon, soak it in 20% honey for 3 hours, and drain. Freeze drying: adjust the temperature of the freeze drying equipment drying box plate layer and the product to -50°C, so that the liquid component in the deer tendon reaches the eutectic point, freeze dry for 2 hours; adjust the temperature to -55°C and turn on the vacuum pump, freeze dry for 0.5 hours; then slowly heat the plate layer until the pressure in the drying box reaches 32Pa, and maintain the effective pressure for 6 hours; after the moisture content is determined to be qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 70-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain deer tendon powder.

[0060] 2. Weigh the washed and dried mushrooms and freeze-dry them: adjust the temperature of the freeze-drying equipment drying box plate layer and the product to -50°C, so that the liquid component in the mushrooms reaches the eutectic point, and freeze-dry for 1 hour; then adjust the temperature to -55°C and turn on the vacuum pump, freeze-dry for 0.5 hour; then slowly heat the plate layer until the pressure in the drying box reaches 32Pa, and maintain the effective pressure for 4 hours; after the moisture content is qualified (the mass fraction of moisture is ≤18%), grind it into powder, pass through a 70-mesh sieve, and repeat the above-mentioned crushing and sieving steps for the large particles that do not meet the particle size requirements, and mix the obtained powders together to obtain mushroom powder.

[0061] 3. Weigh millet, fry it in a drum frying pan at 160°C for 5 minutes, crush it into powder, pass it through a 50-mesh sieve, and repeat the above crushing and sieving steps for the large particles that do not meet the particle size requirements. Then, mix the obtained powders together to obtain millet flour.

[0062] 4. Weigh the washed and dried wolfberries, freeze-dry them (the specific steps are the same as 2. of the present embodiment), grind them into powder after the moisture content is determined to be qualified (the mass fraction of moisture is ≤18%) after freeze-drying, and pass through a 70-mesh sieve. After sieving the large particles that do not meet the particle size requirements, repeat the above-mentioned crushing and sieving steps, and mix the obtained powders together to obtain wolfberry powder.

[0063] 5. Mix the deer hoof tendon powder, Taiwanese mushroom powder, millet powder and wolfberry powder together and stir evenly.

[0064] 6. Add sodium alginate and stir evenly again to obtain a deer tendon composition.

[0065] Comparative Example 1

[0066] Weigh 50 g of fresh deer tendon after purification and prepare deer tendon powder by the method of step 1 of Example 1.

[0067] Test Example 1

[0068] Antioxidant and Anti-Aging Research

[0069] All data in this test case are expressed as mean ± standard deviation. SPSS22.0 software was used for statistical analysis. Normal distribution and variance homogeneity tests were performed before intergroup comparisons in each experimental group. One-way analysis of variance was used for intergroup comparisons. P < 0.05 represented a significant difference, and P < 0.01 represented an extremely significant difference.

[0070] 1. Materials and Methods

[0071] 1.1 Materials: MDA kit, SOD kit, GSH-Px kit, protein concentration assay kit, mouse hyaluronic acid (HA) ELISA kit, mouse hydroxyproline (Hyp) ELISA kit.

[0072] 1.2 Methods

[0073] 1.2.1 Grouping and modeling methods

[0074] 40 ICR mice were randomly divided into 4 groups (10 in each group) according to body weight, namely, control group, model group, deer tendon composition group, and deer tendon group. After adaptive feeding for 1 week, the control group was intraperitoneally injected with normal saline every day, and the other groups were intraperitoneally injected with 120 mg / kg of D-galactose every day for aging modeling, and each group was injected continuously for 8 weeks. At the same time as modeling, mice were gavaged at a volume of 0.1 mL / 10 g. The model group and the control group were gavaged with normal saline, and the deer tendon composition group and the deer tendon group were gavaged with the same volume of samples, and the gavage dose was 10 g / kg; among them, the deer tendon composition group was gavaged with the sample prepared in Example 1 and boiled water, and the deer tendon group was gavaged with the sample prepared in Comparative Example 1 and boiled water. Each group of mice was gavaged once a day for 8 consecutive weeks. After the last administration, the mice were fasted but not water-restricted. Blood was collected from the orbital venous plexus after isoflurane anesthesia on the 2nd day, and then the mice were killed by cervical dislocation. The back skin tissues of mice in each group were collected under aseptic conditions, marked and quickly stored in liquid nitrogen.

[0075] 1.2.2 Determination of mouse skin moisture content

[0076] The moisture content was determined in accordance with GB 5009.3-2016 "National Food Safety Standard - Determination of Moisture in Food". The mouse skin was accurately weighed and recorded, and then placed in a 100°C constant temperature drying oven. The drying time should not be too long each time, and the skin weight was weighed and recorded once every 30 minutes until the difference in weight was less than 2 mg for two consecutive times. The last weighed skin weight was selected as the sample weight after drying, and the final moisture content was calculated using the following formula.

[0077] X=(m2-m1) / m2×100%

[0078] Where: X is the moisture content (%); m2 is the mass of the sample before drying (g); m1 is the mass of the sample after drying (g).

[0079] The results are shown in Table 1 below. The skin moisture content of the aging model group was significantly reduced compared with the control group, indicating that aging can significantly reduce the skin moisture content, and further verifying that the D-galactose-induced aging mouse model was successfully established. Deer tendon and deer tendon composition can significantly increase the skin moisture content of mice, among which the deer tendon composition group is significantly different from the model group (P<0.01), and the deer tendon composition group is significantly different from the deer tendon group (P<0.05), indicating that the deer hoof composition prepared by the present invention can increase the skin moisture content.

[0080] Table 1 Effect of deer tendon composition on skin moisture content of mice

[0081] Group Skin moisture content% Control group 74.15±5.34 Model Group 51.23±6.78* Deer tendon combination group 75.34±7.24# Deer tendon group 62.87±5.45##

[0082] Note: * indicates P<0.05 compared with the control group, # indicates P<0.01 compared with the model group, ## indicates P<0.05 compared with the deer tendon combination group.

[0083] 1.2.3 Determination of protein content in mouse skin

[0084] Weigh a certain amount of skin tissue, add physiological saline and mix, centrifuge at 4000r / min for 1min, take the supernatant and prepare skin tissue homogenate. Then follow the instructions of the protein concentration determination kit, draw a standard curve with protein concentration as the horizontal axis and absorbance as the vertical axis, and calculate the protein content of the sample to be tested.

[0085] The results are shown in Table 2. Compared with the control group, the protein content of the skin of the aging model group was significantly reduced, indicating that aging can reduce the protein content in the skin. Compared with the aging model group, the protein content of the skin of mice in each experimental group increased, among which the deer tendon composition group had the largest increase in skin protein content, which was significantly different from the aging model group (P<0.01), and the deer tendon composition group had a significant difference from the deer tendon group (P<0.05). The above results show that the deer tendon composition prepared by the present invention helps to increase the content of skin protein.

[0086] Table 2 Effect of deer tendon composition on protein content in mouse skin

[0087] Group Skin protein content% Control group 6.25±0.34 Model Group 3.34±0.24* Deer tendon combination group 5.23±0.38# Deer tendon group 4.87±0.45##

[0088] Note: * indicates P<0.05 compared with the control group, # indicates P<0.01 compared with the model group, ## indicates P<0.05 compared with the deer tendon combination group.

[0089] 1.2.4 Detection of Hydroxyproline and Hyaluronic Acid Content

[0090] Hydroxyproline is mainly found in collagen, rarely in elastin, and not in other proteins, so the content of hydroxyproline can indirectly reflect the amount of collagen. Hyaluronic acid mainly plays a moisturizing role in the skin. It is the best skin moisturizing substance found in nature so far and is known as the most ideal moisturizing factor. At the same time, the content of collagen and hyaluronic acid are closely related. The reduction of hyaluronic acid will directly lead to a decrease in the collagen content in the skin, thereby causing skin aging. Therefore, the content of hydroxyproline and hyaluronic acid is measured.

[0091] Take about 1cm of skin from the back 2 , remove hair and fat, defat with acetone-ether in a mass ratio of 1:1, chop and air-dry. Accurately weigh 20 mg and determine the content of hydroxyproline and hyaluronic acid according to the instructions of the kit.

[0092] The results are shown in Table 3. The hydroxyproline and hyaluronic acid contents in the aging model group were significantly lower than those in the control group (P<0.01), indicating that aging can reduce the content of hydroxyproline and hyaluronic acid in the skin. Compared with the aging model group, the hydroxyproline and hyaluronic acid contents in the skin of mice in the deer tendon composition group were significantly increased (P<0.01), and the difference between the deer tendon composition group and the deer tendon group was extremely significant (P<0.05). The above results show that the deer tendon composition prepared by the present invention can effectively increase the content of hydroxyproline and hyaluronic acid in the skin, and can effectively improve the collagen binding capacity of aging skin.

[0093] Table 3 Effect of deer tendon composition on hydroxyproline and hyaluronic acid content in mouse skin

[0094]

[0095]

[0096] Note: * indicates P<0.01 compared with the control group, # indicates P<0.01 compared with the model group, ## indicates P<0.05 compared with the deer tendon combination group.

[0097] 1.2.5 Determination of antioxidant indexes in mouse serum and skin

[0098] 1.2.5.1 Determination of mouse serum antioxidant indexes

[0099] After blood was collected from the eyeball, it was allowed to stand for 30 minutes and centrifuged at 3000 r / min for 15 minutes. The supernatant was taken and the MDA content and SOD and GSH-Px activities in the serum of each group of mice were determined according to the instructions of the kit.

[0100] The results are shown in Table 4. The activities of SOD and GSH-Px in the serum of mice in the aging model group were significantly lower than those in the control group (P<0.01), while the content of MDA was significantly higher than that in the control group (P<0.01). The activities of SOD and GSH-Px in the serum of mice in the deer tendon group and the deer tendon composition group were significantly increased, and the content of MDA was significantly reduced. Among them, the deer tendon composition group had the best effect, which was significantly different from the aging model group (P<0.01), and the deer tendon composition group was significantly different from the deer tendon group (P<0.05). The above results show that the deer tendon composition prepared by the present invention can effectively repair oxidative damage caused by aging.

[0101] Table 4 Effect of deer tendon composition on antioxidant indexes in mouse serum

[0102] Group SOD GSH-Px MDA Control group 65.2±2.34 73.7±4.43 5.7±1.06 Model Group 26.8±0.52* 35.8±2.64* 11.3±1.43* Deer tendon combination group 55.6±2.8# 64.5±4.24# 6.5±0.72# Deer tendon group 42.8±3.45## 54.3±3.52## 8.1±0.42##

[0103] Note: * indicates P<0.01 compared with the control group, # indicates P<0.01 compared with the model group, ## indicates P<0.05 compared with the deer tendon combination group.

[0104] 1.2.5.2 Determination of antioxidant indexes in mouse skin

[0105] When studying the anti-aging effect and testing the serum antioxidant index, the antioxidant index of skin tissue homogenate is also the key to judging the effect of improving aging. Accurately weigh the mouse skin mass, use physiological saline as the solvent, centrifuge at 4000r / min for 1min, take the supernatant, and prepare the skin tissue homogenate. According to the instructions of the relevant kit, the MDA content and SOD and GSH-Px activities in the skin tissue homogenate of each group of mice were measured.

[0106] The results are shown in Table 5. The activities of SOD and GSH-Px in the skin tissue homogenate of the mice in the aging model group were significantly lower than those in the control group (P<0.01), while the content of MDA was significantly higher than that in the control group (P<0.01). Compared with the aging model group, the activities of SOD and GSH-Px in the skin of mice in each experimental group were significantly increased and the content of MDA was significantly decreased. Among them, the freeze-dried deer tendon combination group had the best effect, which was significantly different from the aging model group (P<0.01). The deer tendon combination group was significantly different from the deer tendon group (P<0.05). The above results show that the deer tendon combination prepared by the present invention can effectively restore oxidative damage to the skin.

[0107] Table 5 Effect of deer tendon composition on antioxidant indexes in mouse skin

[0108]

[0109]

[0110] Note: * indicates P<0.01 compared with the control group, # indicates P<0.01 compared with the model group, ## indicates P<0.05 compared with the deer tendon combination group.

[0111] Test Example 2

[0112] Research on improving skin elasticity and beautifying the skin

[0113] The result data in this test case were statistically analyzed using statistical methods, and the statistical analysis software was SPSS 22.0. If the values ​​were normally distributed, the t-test method was used for statistical analysis; if the values ​​were non-normally distributed, the rank sum test method was used for statistical analysis; the statistical methods all used two-tailed tests, and the test level α = 0.05.

[0114] 1.1 Materials and methods

[0115] 1.1.1 General Information

[0116] This study selected healthy subjects who were admitted to the Department of Plastic Surgery, Central Hospital Affiliated to Changchun University of Chinese Medicine from May 16, 2023 to August 16, 2023. A total of 40 subjects were planned to be enrolled, including 34 females and 6 males, and 38 subjects were completed, including 33 females and 5 males. The number of subjects analyzed was 38, aged 27-48 years old, with an average age of (36.1±3.9) years old.

[0117] 1.1.2 Inclusion criteria

[0118] (1) Age range: 25-50 years old, regardless of gender.

[0119] (2) Select crow's feet level 2-4. The severity of crow's feet is divided into 4 levels, with level 1 being the mildest: visible even when smiling; level 2 being slightly severe: fine lines can be seen even when not smiling; level 3 being severe: deep crow's feet even when not smiling; and level 4 being the most severe: trident lines appear at the corners of the eyes.

[0120] (3)Be in good health.

[0121] (4) During the trial, the participants agree not to use any cosmetics, drugs or health products that may affect the results.

[0122] 1.1.3 Exclusion criteria

[0123] (1) Patients who have used antihistamines in the past week or immunosuppressants or immunomodulatory biological agents and small molecule drugs in the past month;

[0124] (2) Those who had used any anti-inflammatory drugs in the test area within the past two months;

[0125] (3) Subjects with inflammatory skin diseases that have not been clinically cured;

[0126] (4) Patients with insulin-dependent diabetes mellitus;

[0127] (5) Those who have had an allergic disease attack or treatment or used anti-inflammatory drugs such as hormones in the past month;

[0128] (6) Those with serious systemic health problems such as cancer;

[0129] (7) Patients with immunodeficiency or autoimmune diseases;

[0130] (8) Infants, young children, pregnant women and breastfeeding women;

[0131] (9) The judge who considers that scars, pigmentation, atrophy, port-wine stains or other blemishes on the skin to be tested may affect the test results;

[0132] (10) Those with highly sensitive constitutions;

[0133] (11) People who are allergic to fish and its products;

[0134] (12) Those who are not willing to participate or cannot complete the required content according to the trial requirements;

[0135] (13) Other persons who are deemed unsuitable to participate in the experiment by the researchers.

[0136] 1.1.4 Detection

[0137] 1.1.4.1 Skin physiological index detection

[0138] The subjects all took the deer tendon composition prepared in Example 1, starting from the first day of the experiment, twice a day, 10 g each time, for 12 weeks. The skin surface texture testing system VisioScanVC20plus was used to measure skin-related physiological indicators.

[0139] The skin indices, evaluation methods and time points tested in the trial are summarized in Table 6. The testing time was before the start of the trial and at the 4th, 8th and 12th weeks of the trial.

[0140] Table 6 Skin test parameters

[0141] Serial number parameter Evaluation method Test site 1 Percentage of wrinkle area at canthus Visia+Image-ProPlus Corner of the eye 2 Skin smoothness VC20Plus cheek 3 Skin elasticity (R2, F4) Cutometer MPA580 cheek 4 Skin glossiness GlossymeterGL200 cheek 5 Skin Melanin MexameterMX18 cheek 6 Skin b value ColorimeterCL400 cheek 7 Water content of skin stratum corneum CorneometerCM825 cheek

[0142] Results: A total of 40 cases were enrolled, of which 38 completed all 12 weeks of trials and follow-up. The specific results are shown in Table 7. The moisture content in the stratum corneum of the skin was measured using a stratum corneum moisture meter. The higher the test value, the higher the moisture content of the stratum corneum of the skin. The results showed that the moisture content of the stratum corneum of the skin of the subjects increased significantly after taking the deer tendon composition. After 12 weeks, the moisture content increased by 21.5% compared with before using this product. The higher the skin gloss test value, the better the skin gloss. During the use period, the skin gloss of the subjects increased by up to 30.2% (week 12). The skin color test probe measures the yellowness of the skin (b value). The lower the b value, the better the improvement of skin yellowness. The maximum decrease in yellowness during the use of the product is 10.2% (week 12). The pigment test probe tests the changes in skin melanin. The lower the value, the lower the melanin content and the whiter the skin. During the trial period, the test value continued to decrease, with a maximum decrease of 22.9%. The skin elasticity tester is used to test changes in skin elasticity. The most common evaluation reference values ​​are R2 and F4. The R2 value is considered to be the parameter value that is closest to the true state of skin elasticity. The smaller the F4 value, the tighter the skin. During the trial period, the F4 value continued to decrease, with the maximum decrease of 25.8%; the R2 value continued to increase, with the maximum increase of 13%. Visia combined with the image analysis software Image-Pro Plus to analyze the changes in eye wrinkles. The smaller the area ratio, the better. During the use period, this indicator continued to decrease, and the average decrease reached 29.3% in the 12th week.

[0143] Table 7 Changes of skin indexes

[0144]

[0145] 1.1.4.2 Self-evaluation by subjects

[0146] The subjects conducted self-evaluations after using the product for 4, 8, and 12 weeks, and evaluated the 10 descriptions in Table 8 respectively.

[0147] Table 8 Self-evaluation of subjects

[0148]

[0149] The results are shown in Table 9. The subjects had a good subjective experience and the favorable rate gradually increased as the test progressed, which was consistent with the objective skin detection indicators.

[0150] Table 9 Self-evaluation of subjects

[0151]

[0152]

[0153] 1.1.4.3 Safety evaluation

[0154] The product safety is evaluated with reference to the human use test skin grading standards specified in the "Technical Specifications for Safety of Cosmetics 2023 Edition". The reaction grades are: 0, no reaction; 1, weak erythema; 2, erythema, infiltration, papules; 3, erythema, edema, papules, blisters; 4, erythema, edema, bullae.

[0155] The subjects used the product for 12 consecutive weeks. Any adverse events that occurred during the use of the deer tendon composition by observing, questioning, examining and recording the results showed that no adverse skin reactions were observed during the test period.

[0156] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A deer tendon composition, characterized in that: The invention comprises the following raw materials in parts by weight: 50-60 parts of deer tendons, 15-25 parts of Taiwan mushrooms, 6-10 parts of wolfberries, 10-20 parts of millets and 1-3 parts of sodium alginate.

2. The deer tendon composition according to claim 1, characterized in that The invention comprises the following raw materials in parts by weight: 53-57 parts of deer tendons, 18-22 parts of Taiwan mushrooms, 7-9 parts of wolfberries, 13-17 parts of millets and 1.5-2.5 parts of sodium alginate.

3. The method for preparing the deer tendon composition according to any one of claims 1 to 2, characterized in that: The following steps are involved: Deer tendon is soaked in honey, freeze-dried, crushed and sieved to obtain deer tendon powder; The mushrooms are freeze-dried, crushed and sieved to obtain mushroom powder; The millet is roasted, crushed and sieved to obtain millet flour; Lycium barbarum is freeze-dried, crushed and sieved to obtain Lycium barbarum powder; Deer tendon powder, Taiwan mushroom powder, millet powder, wolfberry powder and sodium alginate are mixed to obtain a deer tendon composite.

4. The preparation method according to claim 3, characterized in that: The honey soaking time is 1 to 3 hours.

5. The preparation method according to claim 3, characterized in that: The freeze-drying of deer tendons includes the following steps: freezing the deer tendons soaked in honey at -40°C to -50°C for 2 to 4 hours; then vacuum freezing at -45°C to -55°C for 0.5 to 2.5 hours; and then heating until the pressure reaches 28 to 32 Pa and maintaining for 6 to 8 hours.

6. The preparation method according to claim 3, characterized in that: The freeze drying of Agaricus oleraceus and wolfberry comprises the following steps: freezing Agaricus oleraceus or wolfberry at -40-50℃ for 1-3h; then vacuum freezing at -45-55℃ for 0.5-1.5h; and then heating until the pressure reaches 28-32Pa and maintaining for 4-6h.

7. The preparation method according to claim 3, characterized in that: The millet is roasted at 140-160° C. for 3-5 minutes.

8. The preparation method according to claim 3, characterized in that: The sieving is that the deer tendons, Taiwan mushrooms or wolfberries are sieved through a 40-70 mesh sieve, and the millet is sieved through a 40-50 mesh sieve.

9. The preparation method according to claim 3, characterized in that: The mixing is to mix deer tendon powder, Taiwan mushroom powder, millet powder, wolfberry powder and then mix with sodium alginate.

10. Use of the deer tendon composition according to any one of claims 1 to 2 or the preparation method according to any one of claims 3 to 9 in the preparation of anti-aging, antioxidant and / or beauty products.

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