Deer sinew peptide composition for preventing osteoporosis and preparation method thereof

Abstract

The invention discloses a deer sinew peptide composition for preventing osteoporosis and a preparation method thereof, and relates to the technical field of nutritional compositions.The deer sinew peptide composition specifically comprises, by mass, 30-50 parts of deer sinew peptide, 10-20 parts of deer collagen peptide, 10-20 parts of non-denatured II type collagen, 5-15 parts of cistanche deserticola extract, 0.001-0.01 part of vitamin D, 10-20 parts of calcium and 5-10 parts of magnesium. After rat experiments, the deer sinew peptide composition can effectively prevent osteoporosis, and meanwhile, the components of the deer sinew peptide composition are good in synergistic effect, high in safety and suitable for being taken for a long time.

Description

Technical Field

[0001] The invention relates to the technical field of nutritional compositions, in particular to a deer tendon peptide composition for preventing osteoporosis and a preparation method thereof. Background Art

[0002] Osteoporosis is a systemic bone disease characterized by low bone mass and damage to the microstructure of bone tissue, which leads to increased bone brittleness and increased risk of fractures. It is a chronic disease caused by multiple factors and usually has no special clinical manifestations before a fracture occurs. Osteoporosis is more common in elderly men and postmenopausal women. With the increase of the elderly population in my country, its incidence rate is on the rise. It is a health problem worthy of attention in my country and even around the world.

[0003] However, the drugs commonly used to treat or prevent osteoporosis have certain limitations: bone formation promoters such as parathyroid hormone drugs may cause strong bone resorption, hypercalcemia, arrhythmia, etc. when used for a long time; bone resorption inhibitors such as bisphosphonates may cause nephrotoxicity and mandibular osteonecrosis; supplements such as calcium drugs have limited absorption and utilization, and poor treatment and prevention effects. Therefore, it is important to find a safe and effective alternative for preventing osteoporosis.

[0004] Deer tendon peptide is a bioactive peptide extracted from deer tendons. It is rich in various amino acids, has high biological activity and nutritional value, has broad application prospects, and can be used to increase bone density. However, there is relatively little research and application of compositions based on deer tendon peptide as the main ingredient to prevent osteoporosis, and the composition and ratio of the composition need to be studied.

[0005] In summary, in order to solve the above problems, it is of great significance to develop a deer tendon peptide composition and a preparation method thereof that can effectively prevent osteoporosis, has good synergistic effects of ingredients, is highly safe and is suitable for long-term use. Summary of the invention

[0006] The purpose of the present invention is to provide a deer tendon peptide composition for preventing osteoporosis and a preparation method thereof, so as to solve the problems raised in the prior art.

[0007] To achieve the above object, the present invention provides the following technical solutions:

[0008] A deer tendon peptide composition for preventing osteoporosis comprises the following substances, measured by mass: 30 to 50 parts of deer tendon peptide, 10 to 20 parts of deer collagen peptide, 10 to 20 parts of non-denatured type II collagen, 5 to 15 parts of Cistanche deserticola extract, 0.001 to 0.01 parts of vitamin D, 10 to 20 parts of calcium, and 5 to 10 parts of magnesium.

[0009] Preferably, the preparation method of the deer tendon peptide comprises the following steps: S1: taking deer tendon, crushing it, adding pure water, adjusting the pH to 6-8, adding protease for enzymatic hydrolysis, heating to inactivate the enzyme after completion, centrifuging to obtain a supernatant; S2: separating the supernatant through an ultrafiltration membrane, freeze-drying, and obtaining deer tendon peptide.

[0010] Preferably, the mass ratio of the deer tendon, pure water and protease is 1:5-10:0.02-0.03; and the protease includes one of papain and trypsin.

[0011] In which, during the enzymatic hydrolysis process, the temperature is 40-60°C and the time is 2-6 hours; during the heating process of enzyme inactivation, the temperature is 80-100°C and the time is 10-30 minutes; during the centrifugation process, the speed is 8000-12000rpm and the time is 15-25 minutes; during the ultrafiltration membrane separation process, the retention molecular weight is 3000-10000 Daltons.

[0012] Preferably, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing, adding ethanol solution, refluxing extraction, filtering, and concentrating the filtrate under reduced pressure until there is no alcohol taste, so as to obtain the Cistanche deserticola extract.

[0013] Preferably, the mass ratio of the Cistanche deserticola to the ethanol solution is 1:5-10; and the volume fraction of the ethanol solution is 50-70%.

[0014] Wherein, the time during the reflux extraction process is 2.5 to 3.5 hours.

[0015] Preferably, the deer tendon peptide composition of the present invention can be added with other nutrients or auxiliary materials, such as vitamin K, zinc, etc., as needed to further enhance its efficacy in preventing osteoporosis or improve its taste, stability and other properties, but corresponding experimental studies need to be carried out when adding to verify its safety and effectiveness.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] (1) Deer tendon peptide and deer collagen peptide are rich in various amino acids and active groups, which can promote the proliferation and differentiation of osteoblasts, inhibit the activity of osteoclasts, thereby increasing bone density, and play a major role in the osteoporosis prevention composition; non-denatured type II collagen is mainly extracted from animal cartilage tissue, which can maintain the integrity of cartilage tissue, stimulate the growth and redifferentiation of chondrocytes, and can increase the content and quality of bone collagen, improve the biomechanical properties of bones, improve joint function, effectively reduce the incidence of refracture, and increase joint synovial fluid and improve osteoarthritis; the combination of the three can synergistically promote bone health and prevent osteoporosis. Cistanche deserticola is a traditional commonly used Chinese medicinal material. Its extract can promote osteoblast bone formation BMP2 (bone morphogenetic protein 2), accelerate the proliferation of bone marrow mesenchymal stem cells, and promote osteoblast synthesis enzyme ALP (serum alkaline phosphate), thereby regulating the body's endocrine system and promoting calcium absorption. It can synergize with vitamin D to further enhance the calcium absorption effect. Calcium and magnesium are important components of bones, and together they maintain the normal structure and physiological function of bones.

[0018] (2) The raw materials of the composition provided by the present invention are mostly derived from natural biological resources, and are extracted and processed in a reasonable and specific manner, and do not contain harmful chemicals. Compared with some chemical synthetic drugs, they have fewer side effects and are more safe, and are suitable for long-term use to prevent osteoporosis. DETAILED DESCRIPTION

[0019] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in the field without making any creative work shall fall within the scope of protection of the present invention.

[0020] It should be noted that there is no special restriction on the purchase manufacturers of all the raw materials involved in the present invention, which exemplarily include: papain, trypsin, non-denatured type II collagen, calcium, and magnesium are provided by Shaanxi Tianzeyuan Biotechnology Co., Ltd.; deer collagen peptide is provided by Changchun Luxiang Huatai Biotechnology Co., Ltd.; deer tendons and Cistanche deserticola are both commercially available.

[0021] Embodiment 1:

[0022] Step 1: Preparation of deer tendon peptide: S1: Take 100g of deer tendon, grind it, add 500g of pure water, adjust the pH to 7, add 2g of papain, enzymolyze at 50℃ for 4h, inactivate the enzyme at 90℃ for 20min after the enzymolysis, centrifuge at 10000rpm for 20min to obtain the supernatant; S2: separate the supernatant through an ultrafiltration membrane with a molecular weight cutoff of 5000 Daltons, freeze-dry, and obtain deer tendon peptide.

[0023] Step 2: Preparation of Cistanche deserticola extract: Take 50g of Cistanche deserticola, wash and crush it, add 250g of 60% ethanol solution by volume, reflux extract for 3h, filter, and concentrate the filtrate under reduced pressure until there is no alcohol taste, to obtain the Cistanche deserticola extract.

[0024] Step 3: Preparation of deer tendon peptide composition: weigh 40g of deer tendon peptide, 15g of deer collagen peptide, 15g of non-denatured type II collagen, 10g of Cistanche extract, 0.005g of vitamin D, 15g of calcium, and 8g of magnesium, mix well, and make capsules, each capsule containing 0.5g of deer tendon peptide composition.

[0025] Embodiment 2:

[0026] Step 1: Preparation of deer tendon peptide: S1: Take 100g of deer tendon, grind it, add 600g of pure water, adjust the pH to 6.5, add 3g of trypsin, enzymolyze at 45°C for 5h, inactivate the enzyme at 95°C for 15min, centrifuge at 12000rpm for 15min to obtain the supernatant; S2: separate the supernatant through an ultrafiltration membrane with a molecular weight cutoff of 8000 Daltons, freeze-dry, and obtain deer tendon peptide.

[0027] Step 2: Preparation of Cistanche deserticola extract: Take 60g of Cistanche deserticola, wash and crush it, add 300g of 55% ethanol solution by volume, reflux extract for 2.5h, filter, and concentrate the filtrate under reduced pressure until there is no alcohol taste, to obtain the Cistanche deserticola extract.

[0028] Step 3: Preparation of deer tendon peptide composition: weigh 35g of deer tendon peptide, 18g of deer collagen peptide, 12g of non-denatured type II collagen, 12g of Cistanche extract, 0.008g of vitamin D, 18g of calcium, and 6g of magnesium, mix well, and make an oral solution containing 1g of the composition per 10ml.

[0029] Embodiment 3:

[0030] Step 1: Preparation of deer tendon peptide: S1: Take 100g of deer tendon, grind it, add 800g of pure water, adjust the pH to 7.5, add 2.5g of papain, and perform enzymolysis at 55°C for 3h. After the enzymolysis, inactivate the enzyme at 100°C for 10min, centrifuge at 8000rpm for 25min to obtain the supernatant; S2: Separate the supernatant through an ultrafiltration membrane with a molecular weight cutoff of 6000 Daltons, and freeze-dry to obtain deer tendon peptide.

[0031] Step 2: Preparation of Cistanche deserticola extract: Take 40g of Cistanche deserticola, wash and crush it, add 200g of 65% ethanol solution, reflux extraction for 3.5h, filter, and concentrate the filtrate under reduced pressure until there is no alcohol taste, to obtain the Cistanche deserticola extract.

[0032] Step 3: Preparation of deer tendon peptide composition: weigh 45g of deer tendon peptide, 10g of deer collagen peptide, 18g of non-denatured type II collagen, 8g of Cistanche extract, 0.003g of vitamin D, 12g of calcium, and 10g of magnesium, mix well, and make into granules, each bag containing 5g of the composition.

[0033] Performance test: (1) Experimental animals: 3-month-old female SD rats were randomly divided into a normal control group, a model control group, and an experimental group, with 10 rats in each group;

[0034] (2) Instruments and reagents: Instruments: atomic absorption spectrophotometer, bone densitometer (dual-energy X-ray bone densitometer or solid densitometer), animal dissection instruments, 105°C oven; Reagents: reagents required for calcium content determination (nitric acid, perchloric acid, lanthanum oxide, etc.);

[0035] (3) Experimental design and experimental dosage: Model establishment: Except for the normal control group, the other two groups of rats were fed with a low calcium diet (calcium content 0.1%) for 8 weeks to establish an osteoporosis model; Administration method: The normal control group and the model control group were given an equal amount of physiological saline by gavage, and the experimental group was given the deer tendon peptide composition of the present invention by gavage, with a dose of 1.5 g / kg·d, for 8 weeks;

[0036] (4) Observation indicators: Body weight: measured once a week; Femoral dry weight and bone calcium determination: At the end of the experiment, the femurs of both sides of the rats were taken and baked in an oven at 105°C until constant weight, and the bone dry weight was weighed. The left femur at constant weight was digested by atomic absorption spectrometry to determine the bone calcium content; Bone mineral density determination: The left femur was scanned by dual-energy X-ray method to determine the BMD value of the whole femur (g / cm 2 );

[0037] (5) Data processing and result judgment: The experimental data were analyzed by variance analysis. The variance homogeneity test was performed first. If the variance was homogeneous, the F value was calculated. <F 0.05 Conclusion: There is no significant difference in the mean values ​​of each group; F value ≥ F 0.05 , P≤0.05, the pairwise comparison method between the means of multiple experimental groups and a control group was used for statistics; the non-normal or uneven variance data were appropriately transformed, and after meeting the normality or variance requirements, the transformed data were used for statistics; if the variables still did not meet the purpose of normality or variance equality after transformation, the rank sum test was used for statistics; after the data processing was completed, the numerical changes were judged, and the evaluation results are shown in Table 1 (P<0.05):

[0038] Table 1

[0039] project Femoral dry weight Bone calcium content Bone density Example 1 Significant increase Significantly increased Significantly increased Example 2 Increase Elevated Increase Example 3 Increased Significantly increased Increased

[0040] Conclusion: After testing, compared with the model control group, the weight of the rats in the experimental group increased normally, and the values ​​of femoral dry weight, bone calcium content and bone density were significantly improved, which verified the effectiveness of the composition in preventing osteoporosis. At the same time, we also found that after Example 1 was made into capsules, the use effect was the best, which can also provide inspiration for the application of the deer tendon peptide composition.

[0041] In summary, the deer tendon peptide composition prepared by the present invention can effectively prevent osteoporosis, and the components have good synergistic effects, high safety, and is suitable for long-term use.

[0042] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A deer tendon peptide composition for preventing osteoporosis, characterized in that: The invention comprises the following substances, calculated by mass: 30 to 50 parts of deer tendon peptide, 10 to 20 parts of deer collagen peptide, 10 to 20 parts of non-denatured type II collagen, 5 to 15 parts of Cistanche deserticola extract, 0.001 to 0.01 parts of vitamin D, 10 to 20 parts of calcium and 5 to 10 parts of magnesium.

2. The deer tendon peptide composition for preventing osteoporosis according to claim 1, characterized in that: The preparation method of the deer tendon peptide comprises the following steps: S1: Take deer tendons, grind them, add pure water, adjust the pH to 6-8, add protease for enzymolysis, heat to inactivate the enzyme, centrifuge, and obtain the supernatant; S2: Separate the supernatant through an ultrafiltration membrane, freeze-dry, and obtain deer tendon peptide.

3. The deer tendon peptide composition for preventing osteoporosis according to claim 2, characterized in that: The mass ratio of the deer tendon, pure water and protease is 1:5-10:0.02-0.03; the protease includes one of papain and trypsin.

4. The deer tendon peptide composition for preventing osteoporosis according to claim 2, characterized in that: During the enzymatic hydrolysis process, the temperature is 40-60°C and the time is 2-6 hours; during the heating process to inactivate the enzyme, the temperature is 80-100°C and the time is 10-30 minutes; during the centrifugation process, the rotation speed is 8000-12000 rpm and the time is 15-25 minutes; during the ultrafiltration membrane separation process, the retention molecular weight is 3000-10000 Daltons.

5. The deer tendon peptide composition for preventing osteoporosis according to claim 1, characterized in that: The preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing, adding ethanol solution, refluxing extraction, filtering, and concentrating the filtrate under reduced pressure until there is no alcohol taste, so as to obtain the Cistanche deserticola extract.

6. The deer tendon peptide composition for preventing osteoporosis according to claim 5, characterized in that: The mass ratio of the Cistanche deserticola to the ethanol solution is 1:5-10; the volume fraction of the ethanol solution is 50-70%.

7. The deer tendon peptide composition for preventing osteoporosis according to claim 5, characterized in that: The reflux extraction process lasts for 2.5 to 3.5 hours.

8. The composition prepared according to the method for preparing a deer tendon peptide composition for preventing osteoporosis according to any one of claims 1 to 7.