CN113398241A - Bone peptide composition, bone peptide tablet, preparation method and evaluation method - Google Patents
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Abstract
The invention provides a bone peptide composition, a bone peptide tablet, a preparation method and an evaluation method, wherein the bone peptide composition comprises vitamin K2Powder and collagen peptide powder; the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method, the bone collagen peptide powder is bovine bone collagen powder, and the vitamin K is2The component ratio of the powder to the collagen peptide powder is 1: 40. The bone peptide composition provided by the invention has better effects in the aspects of calcium supplement, bone density increase, bone health improvement and osteoporosis prevention.
Description
Technical Field
The invention relates to the technical field of bone health-care medicines, in particular to a bone peptide composition, a bone peptide tablet, a preparation method and an evaluation method.
Background
In the modern society, the incidence rate of osteoporosis is higher and higher, and the osteoporosis becomes a global health problem, and particularly poses serious threats to the health of middle-aged and elderly people.
Among them, various factors cause a decrease in bone density. For example, bone microstructural disruption or increased bone fragility can cause osteoporosis problems. Bone density measurement is currently the accepted "gold standard" in academia for diagnosing osteoporosis, and the degree of osteoporosis can also be estimated. Specifically, a greater bone density indicates better bone mass and bone structure, and increasing bone density is the most effective way to prevent or ameliorate osteoporosis. It is reported that vitamin D and vitamin K2And calcium, which are closely related to bone health. In particular, calcium is an important raw material for constructing bones and teeth. Vitamin D promotes calcium absorption and is a vehicle. Vitamin K2Calcium is carried from the blood into the bone and the three nutrients need to work synergistically to maintain bone health.
The bone peptide is a bone collagen peptide, which is a bone collagen product prepared by taking bone gelatin as a raw material, applying an enzymolysis process and controlling reaction conditions. Clinical studies have shown that: bone peptide and vitamin K2All have effects of preventing and treating osteoporosis and enhancing bone density, bone peptide and vitamin K2The compound can play a role in synergy and plays an important role in the prevention and treatment of osteoporosis. At present, some calcium supplement medicines are also disclosed in the prior art.
For example, prior patent 1 (application number: 201810440537.8) discloses a colostrum mineral salt calcium supplement preparation, which is prepared from the following raw materials in parts by weight: 400-600 parts of milk mineral salt; 40-6 parts of biological calcium carbonate0 part of (C); 200-250 parts of ossein protein; 20-30 parts of colostrum basic protein; 100-150 parts of glucosamine; 30-60 parts of chondroitin sulfate; 30-60 parts of magnesium stearate; vitamin D3 0.5-1.5 parts; vitamin K20.25 to 1.0 portion. However, the composition uses calcium salt as main ingredient, and the collagen is only used as auxiliary material, and the effect of calcium salt is more inclined, the source of the collagen is not described, and the essential effect of the composition is derived from collagen peptide and vitamin K2Compatibility of medicines. The prior patent 2 (application number: 201811176369.2) discloses a vitamin K composition, a calcium balancing preparation comprising the same and application thereof, wherein the vitamin K composition comprises the following components in parts by weight: vitamin K20.01 to 0.1 weight part, 40 to 1200 weight parts of type II collagen and 200 to 1000 weight parts of calcium. Although this patent discloses collagen and vitamin K2The composition has the effects of increasing bone density, maintaining calcium balance of human body and the like, but the hydrolyzed collagen has wide sources and various varieties, and is still lack of guidance in enterprise production and application.
Based on the above description, the existing bone peptide product has the problems of single and blind compatibility, unclear mechanism and effect, and incapability of realizing large-scale mass production and application.
Disclosure of Invention
Based on the above, the invention aims to solve the problems that the existing bone peptide product is single and blind in compatibility, unclear in mechanism and effect and cannot be well applied to large-scale mass production.
The invention provides a bone peptide composition, wherein the bone peptide composition comprises vitamin K2Powder and collagen peptide powder;
wherein the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method, the bone collagen peptide powder is bovine bone collagen powder, and the vitamin K is2The component ratio of the powder to the collagen peptide powder is 1: 40.
The bone peptide composition provided by the invention contains vitamin K2Powder and collagen peptide powder, and collagen peptideThe powder is bovine bone collagen powder with clear material source and vitamin K2The powder and the bone collagen peptide powder have definite proportion and can be produced in batch. Meanwhile, the medicine test experiment shows that the bone peptide composition provided by the invention has better effects on calcium supplement, bone density increase, bone health improvement and osteoporosis prevention.
The bone peptide composition, wherein the vitamin K2The carrier in the powder is maltodextrin, and in the bone peptide composition, the vitamin K2The powder content is more than 40 ug/g.
The invention also provides a bone peptide tablet, wherein the bone peptide tablet is prepared from the bone peptide composition, and the bone peptide tablet comprises:
bone collagen peptide powder, shark cartilage powder, maltodextrin, microcrystalline cellulose, and vitamin K2Powder, magnesium stearate, hydroxypropyl methylcellulose, polyethylene glycol, and talc;
wherein the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method, the bone collagen peptide powder is bovine bone collagen powder, and the vitamin K is2The component ratio of the powder to the collagen peptide powder is 1: 40.
The bone peptide tablet comprises the following components in parts by weight:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
Wherein the sum of the total weight of the hydroxypropyl methyl cellulose, the polyethylene glycol and the talcum powder is 2 parts.
The invention also provides a preparation method of the bone peptide tablet, wherein the method is used for preparing the bone peptide tablet, and the method comprises the following steps:
the method comprises the following steps: mixing the bone collagen peptide powder with the shark cartilage powder, and uniformly stirring to obtain a mixture A;
step two: mixing fructus Hordei GerminatusDextrin, microcrystalline cellulose vitamin K2Uniformly mixing the powder to obtain a mixture B;
step three: uniformly mixing the mixture B with the mixture A, and performing boiling granulation;
step four: sieving the granules obtained in the third step, and finishing the granules to obtain dry granules;
step five: mixing magnesium stearate with the dry granules to obtain a mixture;
step six: tabletting the mixture to obtain plain tablets;
step seven: and adding the coating powder into purified water to prepare a coating solution, and coating the plain tablets by the coating solution to obtain the bone peptide tablets.
The preparation method of the bone peptide tablet comprises the following steps of:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
Wherein the coating powder comprises hydroxypropyl methyl cellulose, polyethylene glycol and talcum powder, and the sum of the corresponding total weight parts is 2 parts.
The preparation method of the bone peptide tablet comprises the step of hydrolyzing bone collagen by a hydrolysis method to obtain bone collagen peptide powder, wherein the bone collagen peptide powder is bovine bone collagen powder.
The preparation method of the bone peptide tablet comprises the following steps of, in the fourth step, sieving the granules in the third step through a 60-mesh sieve to obtain dry granules;
in the fifth step, the mixing time of the magnesium stearate and the dry granules is 5-15 min.
The invention also provides an evaluation method of the bone peptide composition, which is used for evaluating the drug effect of the bone peptide composition and comprises the following steps:
the method comprises the following steps: construction of osteoporosis animal model and oral administration experiment
After monthly healthy female SD rats were acclimatized for 1W, they were randomly divided into 3 groups: control group, model group and experimental group; in the morning, rats in the model group and the experimental group were gavaged with tretinoin suspension at a dose of 120mg/kg, and rats in the control group were gavaged with distilled water at the same amount for 2 w; the rats in the experimental group are gavaged with the bone peptide composition at a dose of 1.025g/kg, and the gavaged amounts of distilled water in the control group and the model group are kept for 4 w;
step two: sample collection
After 4w of intragastric administration, the rats of the control group, the model group and the experimental group are respectively transferred to a metabolism cage, and the urine of the rats is continuously collected for 24 hours and is frozen and stored in an environment of-20 ℃; the next day, carrying out intraperitoneal injection on the rat by utilizing pentobarbital in an amount of 6 mg/kg, cutting the thoracic cavity of the rat after the rat is anesthetized, inserting a needle from the apex of the heart, puncturing the left ventricle, slowly drawing blood, standing overnight at the temperature of 4 ℃, centrifuging at 3000 r/s for 10min, taking serum, and packaging and freezing at the temperature of-20 ℃; after blood is taken through cardiac puncture, the skin of the leg of a rat is cut, the femurs on the left side and the right side of the rat are sequentially taken, soft tissues are removed, and the right femurs are fixed in tissue fixing liquid and stored at the temperature of 4 ℃; wrapping the left femur in gauze soaked with normal saline, and freezing at-80 deg.C;
step three: bone property measurement
Carrying out bone characteristic index measurement on mice of a control group, a model group and an experimental group, wherein the measurement items comprise femur length, femur dry weight, femur index and tibia length;
step four: detection of bone calcium content in blood
Preparing 10 mu g/ml of mixed standard stock solution containing Ca/P, wherein the standard solution series is prepared by diluting standard stock solution step by step, and the medium is 5% Merk-level nitric acid; preparing a 10 mu g/ml rhodium and rhenium standard solution by taking 5 percent Merk-grade nitric acid as a medium, and diluting to 1 mu g/ml to be used as an internal standard solution; the tuning solution is a mixed standard solution (2% nitric acid medium) of lithium, cobalt, yttrium, cerium and thallium with the concentration of 10 ng/ml; weighing 0.05g of femoral sample of each rat, weighing the femoral sample to 50ml of PET plastic bottle by using 5% Merk-level nitric acid, uniformly mixing the femoral sample and the PET plastic bottle, setting a blank group, and selecting an isotope under optimized experimental conditions43Ca, for sample digestion solutionPerforming an analytical test to obtain the bone calcium content;
step five: detection of bone alkaline phosphatase and alkaline phosphatase content in blood
Placing the serum of each group of rats stored in the environment of 20 ℃ below zero on ice to gradually melt, and measuring the concentration of bone alkaline phosphatase and alkaline phosphatase in blood;
step six: determination of hydroxyproline concentration in urine
Placing urine of each group of rats stored in the environment of-20 ℃ on ice to gradually melt, and detecting the concentration of hydroxyproline in the urine by using a hydroxyproline kit; the determination step comprises alkaline hydrolysis, pH adjustment to 6.0-6.8, oxidation of an oxidant, color development of dimethylaminobenzaldehyde, determination of absorbance by using an enzyme-labeling instrument at a wavelength of 550nm, and calculation of the concentration of hydroxyproline in the urine sample to be determined according to the OD value;
step seven: and evaluating the efficacy of the bone peptide composition according to the bone density detection result, the bone characteristic measurement parameters, bone calcium, bone alkaline phosphatase, alkaline phosphatase and hydroxyproline concentration.
In order to make the aforementioned and other objects, features and advantages of the present invention comprehensible, preferred embodiments accompanied with figures are described in detail below.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a flow chart of a method for preparing a bone peptide tablet according to the present invention;
FIG. 2 is a flow chart of the method for evaluating the bone peptide composition according to the present invention.
Detailed Description
To facilitate an understanding of the invention, the invention will now be described more fully with reference to the accompanying drawings. Preferred embodiments of the present invention are shown in the drawings. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The existing bone peptide product has the problems of single and blind compatibility, unclear mechanism and effect and incapability of realizing large-scale batch production and application.
In order to solve the technical problem, the invention provides a bone peptide composition, wherein the bone peptide composition comprises vitamin K2Powder and bone collagen peptide powder.
Wherein the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method. Specifically, in this embodiment, the selected collagen peptide powder is bovine collagen powder. At the same time, the above vitamin K2The component ratio of the powder to the collagen peptide powder is 1: 40.
Here, it is to be noted that vitamin K2The carrier in the powder is maltodextrin. In the above bone peptide composition, vitamin K2The powder content is more than 40 ug/g.
The bone peptide composition provided by the invention contains vitamin K2Powder and bone collagen peptide powder, wherein the bone collagen peptide powder is bovine bone collagen powder with clear material source, and vitamin K2The powder and the bone collagen peptide powder have definite proportion and can be produced in batch. Meanwhile, the medicine test experiments show that the bone peptide composition provided by the invention has better effects on calcium supplement, bone density increase, bone health improvement and osteoporosis prevention。
The invention also provides a bone peptide tablet, wherein the bone peptide tablet is prepared from the bone peptide composition, and the bone peptide tablet comprises:
bone collagen peptide powder, shark cartilage powder, maltodextrin, microcrystalline cellulose, and vitamin K2Powder, magnesium stearate, hydroxypropyl methylcellulose, polyethylene glycol, and talc.
As described above, the collagen peptide powder is obtained by hydrolyzing collagen by a hydrolysis method. In this embodiment, the collagen peptide powder is bovine collagen powder.
Specifically, the bone peptide tablet comprises the following components in parts by weight:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
Wherein the sum of the total weight of the hydroxypropyl methyl cellulose, the polyethylene glycol and the talcum powder is 2 parts.
The invention also provides a preparation method of the bone peptide tablet, wherein the method is used for preparing the bone peptide tablet, and the method comprises the following steps:
the method comprises the following steps: mixing the bone collagen peptide powder and the shark cartilage powder, and uniformly stirring to obtain a mixture A.
Step two: mixing maltodextrin, microcrystalline cellulose and vitamin K2The powders were mixed well to obtain mixture B.
In the step, the added components are respectively as follows in parts by weight:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
It should be noted that the collagen peptide powder is obtained by hydrolyzing collagen by a hydrolysis method, and the collagen peptide powder is bovine collagen powder.
Step three: and uniformly mixing the mixture B with the mixture A, and performing boiling granulation.
Step four: and (4) sieving the granules obtained in the third step, and finishing the granules to obtain dry granules.
In this step, the granules were sieved through a 60 mesh sieve to obtain dry granules.
Step five: magnesium stearate is mixed with the dry granules to obtain a blend.
In the step, the mixing time of the magnesium stearate and the dry granules is 5-15 min.
Step six: and tabletting the mixture to obtain plain tablets.
Step seven: and adding the coating powder into purified water to prepare a coating solution, and coating the plain tablets by the coating solution to obtain the bone peptide tablets.
Wherein the coating powder comprises hydroxypropyl methyl cellulose, polyethylene glycol and talcum powder, and the sum of the corresponding total weight parts is 2 parts.
The prepared bone peptide composition is subjected to animal level efficacy research, and the effects of the bone peptide composition on improving bone density and treating osteoporosis are evaluated.
Specifically, the bone peptide composition is selected as an experimental group, fish skin collagen, pig skin collagen and chicken skin collagen are selected as a control group, and the effects of increasing bone density and preventing and treating osteoporosis of mice of each group are evaluated.
It should be noted that the source information of the raw materials used in the following examples and comparative examples is shown in table 1:
raw materials | Source |
Bone peptide | Bovine bone collagen peptide |
Vitamin K2 powder | Ordinary vitamin K2 powder |
Collagen peptide # 1 | Pigskin collagen peptide |
2# collagen peptide | Fish skin collagen peptide |
3# collagen peptide | Pig bone collagen peptide |
Example 1
In the present invention, a bone peptide composition in which m (vitamin K) is used as an experimental group2) M (bone peptide =1:40, corresponding to example 1.
The evaluation test method set forth in example 1 includes the following steps:
the method comprises the following steps: construction of osteoporosis animal model and oral administration experiment
After monthly healthy female SD rats were acclimatized for 1W, they were randomly divided into 3 groups: control group, model group and experimental group; in the morning, rats in the model group and the experimental group were gavaged with tretinoin suspension at a dose of 120mg/kg, and rats in the control group were gavaged with distilled water at the same amount for 2 w; the rats in the experimental group are gavaged with the bone peptide composition at a dose of 1.025g/kg, and the gavaged amounts of distilled water in the control group and the model group are kept for 4 w;
step two: sample collection
After 4w of intragastric administration, the rats of the control group, the model group and the experimental group are respectively transferred to a metabolism cage, and the urine of the rats is continuously collected for 24 hours and is frozen and stored in an environment of-20 ℃; the next day, carrying out intraperitoneal injection on the rat by utilizing pentobarbital in an amount of 6 mg/kg, cutting the thoracic cavity of the rat after the rat is anesthetized, inserting a needle from the apex of the heart, puncturing the left ventricle, slowly drawing blood, standing overnight at the temperature of 4 ℃, centrifuging at 3000 r/s for 10min, taking serum, and packaging and freezing at the temperature of-20 ℃; after blood is taken through cardiac puncture, the skin of the leg of a rat is cut, the femurs on the left side and the right side of the rat are sequentially taken, soft tissues are removed, and the right femurs are fixed in tissue fixing liquid and stored at the temperature of 4 ℃; wrapping the left femur in gauze soaked with normal saline, and freezing at-80 deg.C;
step three: bone property measurement
Carrying out bone characteristic index measurement on mice of a control group, a model group and an experimental group, wherein the measurement items comprise femur length, femur dry weight, femur index and tibia length;
step four: detection of bone calcium content in blood
Preparing 10 mu g/ml of mixed standard stock solution containing Ca/P, wherein the standard solution series is prepared by diluting standard stock solution step by step, and the medium is 5% Merk-level nitric acid; preparing a 10 mu g/ml rhodium and rhenium standard solution by taking 5 percent Merk-grade nitric acid as a medium, and diluting to 1 mu g/ml to be used as an internal standard solution; the tuning solution is a mixed standard solution (2% nitric acid medium) of lithium, cobalt, yttrium, cerium and thallium with the concentration of 10 ng/ml; weighing 0.05g of femoral sample of each rat, weighing the femoral sample to 50ml of PET plastic bottle by using 5% Merk-level nitric acid, uniformly mixing the femoral sample and the PET plastic bottle, setting a blank group, and selecting an isotope under optimized experimental conditions43Ca, analyzing and testing the sample digestion solution to obtain the bone calcium content;
step five: detection of bone alkaline phosphatase (BALP) and alkaline phosphatase (ALP) content in blood
Placing the serum of each group of rats stored in the environment of 20 ℃ below zero on ice to gradually melt, and measuring the concentration of bone alkaline phosphatase and alkaline phosphatase in blood;
step six: determination of Hydroxyproline (HOP) concentration in urine
Placing urine of each group of rats stored in the environment of-20 ℃ on ice to gradually melt, and detecting the concentration of hydroxyproline in the urine by using a hydroxyproline kit; the determination step comprises alkaline hydrolysis, pH adjustment to 6.0-6.8, oxidation of an oxidant, color development of dimethylaminobenzaldehyde, determination of absorbance by using an enzyme-labeling instrument at a wavelength of 550nm, and calculation of the concentration of hydroxyproline in the urine sample to be determined according to the OD value;
step seven: and evaluating the efficacy of the bone peptide composition according to the bone density detection result, the bone characteristic measurement parameters, bone calcium, bone alkaline phosphatase, alkaline phosphatase and hydroxyproline concentration.
Example 2
The same is true of the experimental group in example 2. Compared with example 1, the difference is only that: in the bone peptide composition, m (vitamin K)2) M (bone peptide) =1: 50.
Comparative example 1
Comparative example 1 corresponds to a control group. Compared with example 1, the difference is only that: after the osteoporosis model was constructed, rats were gavaged with 1g/kg of bone peptide solution daily for 4w in the afternoon.
Comparative example 2
Comparative example 2 corresponds to a control group. Compared with example 1, the difference is only that: after the osteoporosis model was constructed, rats were gavaged with vitamin K at a dose of 25mg/kg every afternoon2Powder, for 4 w.
Comparative example 3
Comparative example 3 corresponds to a control group. Compared with example 1, the difference is only that: the bone peptide in the bone peptide composition is replaced by the collagen peptide # 1.
Comparative example 4
Comparative example 4 corresponds to the control group. Compared with example 1, the difference is only that: the bone peptide in the bone peptide composition is replaced by the # 2 collagen peptide.
Comparative example 5
Comparative example 5 corresponds to a control group. Compared with example 1, the difference is only that: the bone peptide in the bone peptide composition is replaced by the collagen peptide # 3.
Measurement results and analysis:
(1) table 1: bone mineral density test results
Bone Density (g cm)-2) | |
Control group | 250.07±2.55 |
Model set | 186.37±3.69** |
Example 1 | 238.43±2.90**## |
Example 2 | 227.36±1.97**## |
Comparative example 1 | 222.17±7.31**## |
Comparative example 2 | 201.84±2.82**## |
Comparative example 3 | 217.39±2.10**## |
Comparative example 4 | 213.78±2.30**## |
Comparative example 5 | 224.92±1.64**## |
Note: p <0.05, p < 0.01, compared to control; compared with the model group, # p <0.05, # p < 0.01
The results of bone density measurement are shown in Table 1, and compared with the control group, the bone density of the model group rat femur is obviously reduced (p is less than 0.01), which indicates that the osteoporosis modeling is successful. After the other medicaments are perfused, compared with the model group rats, the bone density is obviously increased (p is less than 0.01), and the bone peptide composition (bone peptide and vitamin K)2) The bone density of the corresponding experimental group was higher than that of the other groups, indicating that the bone peptide composition (bone peptide + vitamin K)2) Has more obvious effect on increasing the bone density of rats.
Table 2: measuring meter for bone characteristic index
Femur length (cm) | Dry weight of femur (g) | Stock index | Tibia Length (cm) | Tibia dry weight (g) | Index of tibia | |
Control group | 4.67±0.05 | 0.667±0.009 | 0.143±0.002 | 3.78±0.07 | 0.497±0.006 | 0.131±0.009 |
Model set | 4.11±0.03** | 0.521±0.004** | 0.127±0.002** | 3.29±0.06** | 0.389±0.024** | 0.118±0.004 |
Example 1 | 4.45±0.03**## | 0.647±0.007*## | 0.145±0.006## | 3.56±0.04**## | 0.478±0.022## | 0.134±0.005# |
Example 2 | 4.27±0.04**## | 0.639±0.008*## | 0.149±0.001## | 3.42±0.11* | 0.470±0.036# | 0.137±0.006# |
Comparative example 1 | 4.20±0.04**# | 0.621±0.005**## | 0.147±0.002## | 3.39±0.03** | 0.452±0.050 | 0.133±0.007# |
Comparative example 2 | 4.15±0.04** | 0.575±0.004**## | 0.148±0.001*## | 3.37±0.05** | 0.399±0.029** | 0.118±0.005 |
Comparative example 3 | 4.35±0.02**## | 0.581±0.008**## | 0.134±0.002*# | 3.46±0.07**# | 0.443±0.042 | 0.128±0.002# |
Comparative example 4 | 4.31±0.03**## | 0.604±0.004**## | 0.140±0.001## | 3.42±0.10** | 0.426±0.025** | 0.125±0.004 |
Comparative example 5 | 4.22±0.04**# | 0.618±0.005**## | 0.146±0.002## | 3.38±0.02** | 0.457±0.036# | 0.135±0.005## |
Note: p <0.05, p < 0.01, compared to control; compared with the model group, # p <0.05, # p < 0.01
The measurement of the bone property index is shown in table 2, and it can be seen from table 2 that: the femoral length, femoral dry weight, femoral index, tibial length, tibial dry weight, and tibial index were significantly higher in the control group than in the model group (p < 0.05).
Bone peptide composition (bone peptide + vitamin K)2) The femur length, femur dry weight, femur index, tibia length, tibia dry weight and tibia index of the group are higher than those of the model group, the bone peptide group and the vitamin K2Group 1# collagen + vitamin K2Group, 2# collagen + vitamin K2 group, 3# collagen + vitamin K2Group (p <0.05, p < 0.01), with the lowest index of skeletal properties in the model group.
It can therefore be deduced that: hydrolyzed collagen and vitamin K2Can improve the skeletal characteristics of rats, and bovine bone collagen peptide + vitamin K2The composition has optimal improvement effect.
Table 3: serum bone calcium, BALP, ALP content and urine HOP content
Bone calcium (mg. g-1) | BALP(ng·mL-1) | ALP(U/L) | HOP/(ng·mL-1) | |
Control group | 375.2±22.36 | 9.13±0.23 | 121.86±5.34 | 0.727±0.234 |
Model set | 261.4±13.21** | 4.52±0.12** | 75.63±1.45** | 5.123±0.312** |
Example 1 | 354.7±14.52## | 7.56±0.36**## | 101.31±2.46**## | 2.156±0.065**## |
Example 2 | 337.1±17.41## | 7.52±0.24**## | 96.38±2.31**## | 2.712±0.054**## |
Comparative example 1 | 324.0±11.97*## | 6.98±0.41**## | 86.21±2.44**## | 3.564±0.156**## |
Comparative example 2 | 289.7±9.34**# | 5.36±0.32**# | 80.36±3.01** | 4.145±0.434**# |
Comparative example 3 | 312.2±10.98*## | 7.01±0.33**## | 95.64±1.56**## | 3.213±0.345**## |
Comparative example 4 | 329.5±13.12*## | 7.15±0.47**## | 87.23±1.88**## | 4.012±0.245**## |
Comparative example 5 | 317.6±24.22*# | 7.22±0.38**## | 82.41±3.24**# | 3.891±0.268**## |
Note: p <0.05, p < 0.01, compared to control; compared with the model group, # p <0.05, # p < 0.01
The contents of bone calcium, BALP and ALP in serum and the content of HOP in urine are shown in table 3, and can be known from table three: the retinoic acid induces the osteoporosis of the rat so that the bone calcium content of the rat is remarkably reduced (p)<0.01), rats were supplemented with collagen + vitamin K, as compared with the model group2After the composition, the content of bone calcium is obviously increased (p)<0.05,p<0.01) vitamin K alone2The bone calcium content cannot be increased significantly. Wherein, bovine bone collagen peptide + vitamin K2After treatment, the bone calcium content of rats increased most significantly.
BALP is an enzyme secreted from osteoblasts, and is capable of hydrolyzing inorganic phosphate to reduce its inhibitory effect on bone salt formation, thereby promoting bone formation, and osteoporosis is more likely to occur as the serum BALP concentration is lower. Urine HOP is produced by the degradation of urinary hydroxyproline collagen, and when the blood calcium is reduced, the dissolution and absorption of bones occur, and the excretion of urinary hydroxyproline is increased. The lower the HOP concentration, the higher the bone formation rate. When bone formation is active, osteoblasts secrete a large amount of ALP, one part of ALP participates in bone calcification, and the other part of ALP is released into blood to enhance the activity of enzyme in blood, so that ALP is an index for evaluating the bone formation activity.
As can be seen from Table 3: the retinoic acid induces osteoporosis of rats, so that the BALP and ALP content of the rats is obviously reduced, and the HOP content (p) is obviously increased<0.01). Rats were supplemented with collagen peptide + vitamin K compared to model groups2The composition can increase BALP and ALP content and significantly reduce HOP content<0.05,p <0.01). Wherein, the effect of the treatment by the bovine bone collagen peptide and the vitamin K2 is the best.
In addition, bovine bone collagen peptide or vitamin K is supplemented separately2Has the effect of not supplementing bovine bone collagen peptide and vitamin K2The effect of the composition is obvious. Therefore, to some extent: supplement bovine bone collagen peptide and vitamin K2The composition has good effects on increasing bone formation rate and reducing bone resorption yield.
From the above results, it can be seen that: bovine bone collagen peptide + vitamin K2Has synergistic effect, and has good effects in supplementing calcium, increasing bone density, improving bone health, and preventing osteoporosis.
Finally, it should be noted that: the above-mentioned embodiments are only specific embodiments of the present invention, which are used for illustrating the technical solutions of the present invention and not for limiting the same, and the protection scope of the present invention is not limited thereto, although the present invention is described in detail with reference to the foregoing embodiments, those skilled in the art should understand that: any person skilled in the art can modify or easily conceive the technical solutions described in the foregoing embodiments or equivalent substitutes for some technical features within the technical scope of the present disclosure; such modifications, changes or substitutions do not depart from the spirit and scope of the embodiments of the present invention, and they should be construed as being included therein. Therefore, the protection scope of the present invention shall be subject to the protection scope of the appended claims.
Claims (9)
1. A bone peptide composition comprising vitamin K2Powder and collagen peptide powder;
wherein the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method, the bone collagen peptide powder is bovine bone collagen powder, and the vitamin K is2The component ratio of the powder to the collagen peptide powder is 1: 40.
2. The bone peptide composition of claim 1, wherein the vitamin K is vitamin K2The carrier in the powder is maltodextrin, and in the bone peptide composition, the vitamin K2The powder content is more than 40 ug/g.
3. A bone peptide tablet made from the bone peptide composition of claim 1 or 2 as described above, comprising:
bone collagen peptide powder, shark cartilage powder, maltodextrin, microcrystalline cellulose, and vitamin K2Powder, magnesium stearate, hydroxypropyl methylcellulose, polyethylene glycol, and talc;
wherein the bone collagen peptide powder is prepared by hydrolyzing bone collagen by a hydrolysis method, the bone collagen peptide powder is bovine bone collagen powder, and the vitamin K is2The component ratio of the powder to the collagen peptide powder is 1: 40.
4. The bone peptide tablet of claim 3, wherein the bone peptide tablet comprises the following components in parts by weight:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
Wherein the sum of the total weight of the hydroxypropyl methyl cellulose, the polyethylene glycol and the talcum powder is 2 parts.
5. A process for the preparation of a bone peptide tablet according to claim 3 or 4, comprising the steps of:
the method comprises the following steps: mixing the bone collagen peptide powder with the shark cartilage powder, and uniformly stirring to obtain a mixture A;
step two: mixing maltodextrin, microcrystalline cellulose and vitamin K2Uniformly mixing the powder to obtain a mixture B;
step three: uniformly mixing the mixture B with the mixture A, and performing boiling granulation;
step four: sieving the granules obtained in the third step, and finishing the granules to obtain dry granules;
step five: mixing magnesium stearate with the dry granules to obtain a mixture;
step six: tabletting the mixture to obtain plain tablets;
step seven: and adding the coating powder into purified water to prepare a coating solution, and coating the plain tablets by the coating solution to obtain the bone peptide tablets.
6. The method for preparing the bone peptide tablet as claimed in claim 5, wherein the added components are respectively as follows in parts by weight:
bone collagen peptide powder 80 parts, shark cartilage powder 7 parts
Maltodextrin 4 parts and microcrystalline cellulose 4 parts
Vitamin K2Powder 2 parts magnesium stearate 1 part
Wherein the coating powder comprises hydroxypropyl methyl cellulose, polyethylene glycol and talcum powder, and the sum of the corresponding total weight parts is 2 parts.
7. The method for preparing the bone peptide tablet as claimed in claim 5, wherein the bone collagen peptide powder is obtained by hydrolyzing bone collagen by a hydrolysis method, and the bone collagen peptide powder is bovine bone collagen powder.
8. The method for preparing bone peptide tablets as claimed in claim 5, wherein in the fourth step, the granules in the third step are sieved by a 60-mesh sieve to obtain dry granules;
in the fifth step, the mixing time of the magnesium stearate and the dry granules is 5-15 min.
9. A method for evaluating a bone peptide composition for evaluating the pharmacological effect of the bone peptide composition according to claim 1 or 2, comprising the steps of:
the method comprises the following steps: construction of osteoporosis animal model and oral administration experiment
After monthly healthy female SD rats were acclimatized for 1W, they were randomly divided into 3 groups: control group, model group and experimental group; in the morning, rats in the model group and the experimental group were gavaged with tretinoin suspension at a dose of 120mg/kg, and rats in the control group were gavaged with distilled water at the same amount for 2 w; the rats in the experimental group are gavaged with the bone peptide composition at a dose of 1.025g/kg, and the gavaged amounts of distilled water in the control group and the model group are kept for 4 w;
step two: sample collection
After 4w of intragastric administration, the rats of the control group, the model group and the experimental group are respectively transferred to a metabolism cage, and the urine of the rats is continuously collected for 24 hours and is frozen and stored in an environment of-20 ℃; the next day, carrying out intraperitoneal injection on the rat by utilizing pentobarbital in an amount of 6 mg/kg, cutting the thoracic cavity of the rat after the rat is anesthetized, inserting a needle from the apex of the heart, puncturing the left ventricle, slowly drawing blood, standing overnight at the temperature of 4 ℃, centrifuging at 3000 r/s for 10min, taking serum, and packaging and freezing at the temperature of-20 ℃; after blood is taken through cardiac puncture, the skin of the leg of a rat is cut, the femurs on the left side and the right side of the rat are sequentially taken, soft tissues are removed, and the right femurs are fixed in tissue fixing liquid and stored at the temperature of 4 ℃; wrapping the left femur in gauze soaked with normal saline, and freezing at-80 deg.C;
step three: bone property measurement
Carrying out bone characteristic index measurement on mice of a control group, a model group and an experimental group, wherein the measurement items comprise femur length, femur dry weight, femur index and tibia length;
step four: detection of bone calcium content in blood
Preparing 10 mu g/ml of mixed standard stock solution containing Ca/P, wherein the standard solution series is prepared by diluting standard stock solution step by step, and the medium is 5% Merk-level nitric acid; preparing a 10 mu g/ml rhodium and rhenium standard solution by taking 5 percent Merk-grade nitric acid as a medium, and diluting to 1 mu g/ml to be used as an internal standard solution; the tuning solution is a mixed standard solution (2% nitric acid medium) of lithium, cobalt, yttrium, cerium and thallium with the concentration of 10 ng/ml; weighing 0.05g of femoral sample of each rat, weighing the femoral sample to 50ml of PET plastic bottle by using 5% Merk-level nitric acid, uniformly mixing the femoral sample and the PET plastic bottle, setting a blank group, and selecting an isotope under optimized experimental conditions43Ca, analyzing and testing the sample digestion solution to obtain the bone calcium content;
step five: detection of bone alkaline phosphatase and alkaline phosphatase content in blood
Placing the serum of each group of rats stored in the environment of 20 ℃ below zero on ice to gradually melt, and measuring the concentration of bone alkaline phosphatase and alkaline phosphatase in blood;
step six: determination of hydroxyproline concentration in urine
Placing urine of each group of rats stored in the environment of-20 ℃ on ice to gradually melt, and detecting the concentration of hydroxyproline in the urine by using a hydroxyproline kit; the determination step comprises alkaline hydrolysis, pH adjustment to 6.0-6.8, oxidation of an oxidant, color development of dimethylaminobenzaldehyde, determination of absorbance by using an enzyme-labeling instrument at a wavelength of 550nm, and calculation of the concentration of hydroxyproline in the urine sample to be determined according to the OD value;
step seven: and evaluating the efficacy of the bone peptide composition according to the bone density detection result, the bone characteristic measurement parameters, bone calcium, bone alkaline phosphatase, alkaline phosphatase and hydroxyproline concentration.
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