CN115005352B - Chitosan oligosaccharide and donkey-hide gelatin protein peptide composite and preparation method thereof - Google Patents
Chitosan oligosaccharide and donkey-hide gelatin protein peptide composite and preparation method thereof Download PDFInfo
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- CN115005352B CN115005352B CN202210685260.1A CN202210685260A CN115005352B CN 115005352 B CN115005352 B CN 115005352B CN 202210685260 A CN202210685260 A CN 202210685260A CN 115005352 B CN115005352 B CN 115005352B
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- donkey
- chitosan oligosaccharide
- hide gelatin
- powder
- composition
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Abstract
The invention discloses a chitosan oligosaccharide composite donkey-hide gelatin protein peptide composition and a preparation method thereof, belonging to the technical field of health products and functional foods, and the active ingredients of the composition are prepared from the following raw materials: collagen peptide powder, chitosan oligosaccharide powder, donkey-hide gelatin powder, abelmoschus manihot powder and honey. According to the chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition and the preparation method thereof, provided by the invention, according to the monarch, minister, assistant and guide compatibility theory of traditional Chinese medicine, chitosan oligosaccharide is used as a main component, donkey-hide gelatin, collagen peptide, abelmoschus manihot and honey are used as auxiliary components, the compounded product has stronger functions and better taste, and the defects of poor taste and weak function of a single raw material are overcome. The components in the formula are mutually cooperated, so that the absorption is fast and the bioavailability is high. Particularly, the chitosan oligosaccharide is combined with the donkey-hide gelatin, so that the balance of yin and yang is adjusted, adverse reaction does not occur after the chitosan oligosaccharide and the donkey-hide gelatin are taken, toxic or side effect is avoided, the chitosan oligosaccharide and the donkey-hide gelatin can be taken for a long time as functional food, the physiological balance can be adjusted, and the immunity of a human body is enhanced.
Description
Technical Field
The invention relates to a chitosan oligosaccharide and donkey-hide gelatin protein peptide composite and a preparation method thereof, belonging to the technical field of health products and functional foods.
Background
With the increase of living pressure, the growth of age and the influence of environmental pollution, the immune system of a human body is degraded, so that the immune response of the human body to external specific antigens is reduced, the imbalance of immune functions is generated, the sub-health condition is generated, and various diseases are caused finally. In order to improve the situation, some western medicines with specific curative effects can be taken, but some side effects are easily caused by taking the western medicines in the actual application process. Therefore, in recent years, reasonable consumption of some health care products such as donkey-hide gelatin, collagen and chitosan oligosaccharide has become the choice of more people. However, if the common health care products are taken directly or simply according to respective common dosage, the expected effect is not achieved, for example, donkey-hide gelatin can nourish yin and enrich the blood, but donkey-hide gelatin is negative food, and people with deficiency-cold spleen and stomach eat the health care products, which easily damages intestines and stomach and causes dyspepsia. However, when chitosan oligosaccharide is taken alone, the symptoms of body excessive internal heat such as eye redness, mouth corner ulceration, lip swelling, skin itch and the like are easy to appear.
The above description is included in the technical knowledge of the inventors, and does not necessarily constitute a prior art.
Disclosure of Invention
The invention aims to solve the problems in the prior art and provides a chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition and a preparation method thereof.
The invention adopts the following technical scheme to realize the purpose:
the active ingredients of the chitosan oligosaccharide composite donkey-hide gelatin protein peptide composition are prepared from the following raw materials in parts by weight: 10-30 parts of collagen peptide, 2.5-10 parts of chitosan oligosaccharide, 2.5-10 parts of donkey-hide gelatin, 0.05-0.25 part of abelmoschus manihot and 0.5-5 parts of honey powder.
Optionally, the active ingredients of the composition of chitosan oligosaccharide and donkey-hide gelatin protein peptide provided by the invention are prepared from the following raw materials in parts by weight:
20-30 parts of collagen peptide, 5-10 parts of chitosan oligosaccharide, 5-10 parts of donkey-hide gelatin, 0.02-0.25 part of abelmoschus manihot and 0.5-5 parts of honey powder.
Optionally, the collagen peptide is granular or powdery, the grain size of the granular collagen peptide is 20-30 meshes, the grain size of the powdery collagen peptide is 120-150 meshes, the grain size of the chitosan oligosaccharide is 120-140 meshes, the grain size of the donkey-hide gelatin is 120-150 meshes, the grain size of the abelmoschus manihot is 120-140 meshes, and the grain size of the honey powder is 120-140 meshes.
Optionally, the composition is in the form of any one of powder, granules and solution.
Optionally, the preparation method of the composition of chitosan oligosaccharide and donkey-hide gelatin protein peptide provided by the invention comprises the following steps:
adding the raw materials for forming the active ingredients into a mixer, uniformly mixing, sterilizing and subpackaging to obtain the chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition powder.
Optionally, the preparation method of the chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition provided by the invention can also adopt the following steps: adding the raw materials for forming the active ingredients into a mixer, uniformly mixing, dissolving into pure water while stirring, adding active carbon, clarifying, removing impurities, filtering, sterilizing and filling to obtain the solution of the chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition.
Specifically, the powdered and granular dosage form can be in the form of a solid beverage product, and the solution dosage form can be in the form of a liquid beverage product.
The chitosan oligosaccharide and donkey-hide gelatin protein peptide composite provided by the invention has the following effects of active ingredients:
chitosan oligosaccharide: the oligosaccharide is a micromolecule oligosaccharide with amino degraded by chitosan, has stability to heat, good water solubility, can be directly absorbed by human bodies, has good affinity with the human bodies, can directly participate in the metabolic process of the human bodies, has the functions of regulating acid-base balance, removing in-vivo free radicals, reducing hyperlipidemia and bidirectionally regulating blood pressure and blood sugar, can directly act on hematopoietic stem cells in immune organs of the human bodies, and finally achieves the effect of improving the immunity of the human bodies; the chitosan oligosaccharide has positive charges, and can be neutralized with various acids with negative charges in a human body to form corresponding chitosan oligosaccharide salts to be discharged out of the body. The chitosan oligosaccharide is alkaline oligosaccharide with positive charge, and has antioxidant, antibacterial, antiinflammatory, antiviral, and cancer cell metastasis inhibiting effects.
Collagen peptide: the protein belongs to micromolecule protein, the average molecular weight of the protein is lower than 5000Dalton, the protein is easy to dissolve in water, is stable to acid and heat, can be directly absorbed without being digested, and has high bioavailability; the phagocytic rate and the phagocytic index of macrophages can be increased, the proliferative capacity of lymphocytes can be enhanced, the increase of the number of antibody forming cells can be promoted, and the immune level of an organism can be improved; can eliminate redundant free radicals in vivo and improve the absorption rate of minerals; can increase the activity of glutathione peroxidase, superoxide dismutase and catalase, and reduce the level of lipid peroxide malondialdehyde, thereby playing the role of antioxidation.
E, donkey-hide gelatin: has the effects of nourishing yin, moistening lung, replenishing blood and stopping bleeding, is rich in glycosaminoglycan substances such as protein, lysine, threonine, gelatin, arginine, histidine, trace elements, hyaluronic acid, dermatan sulfate and the like, and has obvious effects on resisting tumors, promoting hematopoiesis and the like.
Abelmoschus manihot: the nutritional ingredients are rich, the contained unsaturated fatty acid can regulate blood fat, and the contained flavone can eliminate free radicals in vivo and reduce cholesterol and blood sugar content; the vitamin E has antioxidant effect; the selenium element can improve cardiovascular and cerebrovascular diseases, resist oxidation, and enhance immunity.
Honey: the honey contains a certain amount of flavonoids, which is helpful for eliminating free radicals in vivo, thereby helping oxidation resistance and aging resistance, and has the functions of resisting bacteria, diminishing inflammation, promoting tissue regeneration, promoting digestion, regulating gastrointestinal function and improving immunity.
Benefits of the present application include, but are not limited to:
according to the composition of chitosan oligosaccharide and donkey-hide gelatin protein peptide and the preparation method thereof, chitosan oligosaccharide is a monarch component in the composition according to the composition theory of traditional Chinese medicine compatibility, monarch, minister, assistant and guide, and plays a main role; the minister component is collagen peptide which assists chitosan oligosaccharide to play a role in synergism; donkey-hide gelatin is an adjuvant component and can restrict the vigor of chitosan oligosaccharide; the abelmoschus manihot is an accessory ingredient and plays a balancing role; the honey is used as messenger component, and has effects of messenger, inducing menstruation, and harmonizing. The chitosan oligosaccharide is used as a main component, and donkey-hide gelatin, collagen peptide, abelmoschus manihot and honey are used as auxiliary components, so that the composite material has stronger function and better taste, and makes up the defects of poor taste and weak function of a single raw material. The components in the formula are mutually cooperated, so that the absorption is fast and the bioavailability is high. Especially, the chitosan oligosaccharide is combined with the donkey-hide gelatin, so that the balance of yin and yang is adjusted, no adverse reaction occurs after the donkey-hide gelatin is taken, no toxic or side effect exists, the chitosan oligosaccharide can be taken as functional food for a long time, the physiological balance can be adjusted, and the immunity of a human body is enhanced.
Detailed Description
The present invention will be described in further detail in the following. It should be noted, however, that the following detailed description merely gives specific operation examples of the present invention by way of example, and the scope of the present invention is not limited thereto. The scope of the invention is limited only by the claims. It will be obvious to those skilled in the art that various other modifications and substitutions can be made to the described embodiments of the invention within the scope of the invention as defined by the claims and still achieve the same technical result and achieve the final technical object of the invention.
Unless otherwise specified, each raw material is commercially available in this specification.
In order to verify the use effect of the composition of chitosan oligosaccharide and donkey-hide gelatin protein peptide provided by the invention, the applicant of the invention carries out the following experimental comparison.
The experimental procedures and results are as follows:
1 materials and methods
1.1 sample
The samples of example 1, comparative example 1 and comparative example 2, which are produced and provided by sunshine shell meile marine biotechnology limited company, are solid powders; specification: each 200 g/bag; storage conditions are as follows: sealing, and storing in shade, dry and dark place at room temperature; the shelf life is 12 months. The production dates of example 1, comparative example 1 and comparative example 2 were all 2022, 3 months and 12 days, and the batch numbers: 20220312. table 1 below shows the specific components of example 1, comparative example 1, and comparative example 2.
TABLE 1 concrete ingredients of example 1, comparative example 1, and comparative example 2
| Formulation components | |
| Example 1 | Collagen peptide powder 118g. 35g of chitosan oligosaccharide powder, 35g of donkey-hide gelatin powder, 2g of abelmoschus manihot powder and 10g of honey powder |
| Comparative example 1 | 118g of collagen peptide powder, 35g of chitosan oligosaccharide powder, 45g of honey powder and 2g of abelmoschus manihot powder |
| Comparative example 2 | 118g of collagen peptide powder, 35g of fructooligosaccharide powder, 35g of donkey-hide gelatin powder, 2g of abelmoschus manihot powder and 10g of honey powder |
1.2 Experimental animals
Selecting Beijing Huafukang biotech GmbH [ license number: 96 female mice of 18 g-22 g SPF KM bred by SCXK (Jing) 2019-0008] are randomly divided into 8 groups after being subjected to animal adaptive feeding observation, each group comprises 12 mice, and each group comprises 4 experimental batches which are 2 experimental batches. The measurement of a delayed type allergy experiment is carried out in an experimental batch; experiment two batches were run for carbon clearance.
Quality certification of experimental animals: no.110322210103078212. The experimental animals are raised in SPF animal room of health food function testing center of applied literature college of Beijing Union university. License number for experimental animals: SYXK (Kyoto) 2017-0038. The maintenance feed is prepared by Beijing Huafukang biotech GmbH [ license number: SCXK (Jing) 2019-0008 ].
1.3 dosage
The experiment is divided into four groups of blank control, example 1, comparative example 1 and comparative example 2, and the tested dose is 30 times of the recommended dose (4 g/d) of a human body, namely 2g/kgBW mice.
Example 1 test article: weighing 5.00g of the sample in example 1, and adding sterile water to the sample to 25.0mL;
comparative example 1 test article: weighing 5.00g of the sample of comparative example 1, and adding sterile water to 25.0mL;
comparative example 2 test article: 5.00g of the sample of comparative example 2 was weighed and sterilized water was added to 25.0mL.
The control group was given an equal amount of sterile water.
The preparation is orally administered once a day, and after continuous gavage for 31d, various indexes are measured.
The gavage dose of the mice was 10mL/kgBW. The blank control group had the same daily gavage dosage as each test group. Maintenance feed was administered to each dose group.
1.4 Main instruments and reagents
The device comprises a T1000 electronic balance (2012003), a BS223S electronic balance (2008007), a BS2202S electronic balance (2014007), a UV2600 ultraviolet visible spectrophotometer (2015003), a SPARK microplate reader (2021001), a carbon dioxide incubator, a centrifuge, a thermostatic water bath, a microscope and a micrometer screw (96099).
A cell counter, 24-well and 96-well flat-bottom cell culture plates, 96-well U-shaped cell culture plate Sheep Red Blood Cells (SRBC), normal saline, hank's solution (pH 7.2-7.4), RPMI1640 culture solution, calf serum, complement (guinea pig serum), SA buffer solution, hb diluent kit, YAC-1 cells, lithium lactate, nitro tetrazole chloride, phenazine dimethyl sulfate, oxidized coenzyme I, 0.2mol/L Tris-HCl buffer solution (pH 8.2), 1-inch NP40, india ink, 0.1% Na 2 CO 3 Solutions, and the like.
1.5 Experimental methods
1.5.1 delayed type allergy (DTH) test (plantar thickening)
Sheep blood was washed 3 times with physiological saline, and each mouse was intraperitoneally injected with 0.2mL of 2% (v/v) SRBC suspension 4d after immunization, and the thickness of the plantar region of the left hind foot was measured. Then 20 μ L of SRBC suspension was injected subcutaneously at the site of measurement (v/v) and the thickness of the plantar portion of the left hind foot was measured 24h after injection. The two measurements are all measured at the same position for three times, and the average value is taken. The degree of DTH is expressed as the difference in plantar thickness before and after the attack. The difference value of the test sample group is obviously higher than that of the control group, and the test result can be judged to be positive.
1.5.2 mouse carbon clearance test
Indian ink (10 mL/10 gBW) diluted 4 times by body weight was injected intravenously from the mouse tail. And immediately timing when the ink is injected.
2 min and 10min after injecting ink, respectively from the inner canthus vein20. Mu.L of blood was taken from the plexus and added to 2.0mL0.1% of Na 2 CO 3 In solution. Measuring optical density value (OD) at 600nm wavelength with spectrophotometer, by 0.1% 2 CO 3 The solution was used as a blank control. The mice were sacrificed and the liver and spleen were taken and blotted with filter paper to dry the surface blood stains of the viscera and weighed. The ability of the mouse to express carbon clearance as phagocytosis index (a) was calculated as follows:
k＝(lgOD1-lgOD2)/(t 2 -t 1 ) A = body weight ÷ (liver weight + spleen weight) × k 1/3
The phagocytosis index of the test sample group is obviously higher than that of the control group, and the positive experimental result can be judged.
1.6 data processing
Data processing was performed with SPSS software. Independent sample T-test was used. The statistical result is divided into two parts, namely 1, the levene test of the homogeneity of the variance; and 2, carrying out t test on the mean value equation. When the uniform variance test result Sig is more than 0.05, the uniform variance is considered, so the mean value equation t is taken to test the Sig value in the first column, and when Sig is less than or equal to 0.05, the statistical results between the two groups are considered to be different; when the variance homogeneous test result Sig is less than or equal to 0.05, the variances are considered to be uneven, so the mean value equation t is taken to test the Sig value in the second column, and when Sig is less than or equal to 0.05, the statistical results between the two groups are considered to be different.
2. Results
2.1. Effect of different test samples on mouse body weight
TABLE 2 initial body weights (x. + -. SD) of the groups of mice
As can be seen from Table 2, the initial body weights of the mice were not significant (P > 0.05) in the formula groups compared with the blank control group. I.e. the initial body weight of the mice was more balanced between the groups.
TABLE 3 Effect on body weight (x. + -. SD) of different test samples
As can be seen from Table 3, after the mice are orally administered with different test samples 31d, the body weight of the mice of each formulation group is not significant (P > 0.05) compared with that of the blank control group. I.e. the different test samples had no adverse effect on the body weight of the mice.
2.2 Effect of different test samples on sheep Red blood cell induced mice DTH (footpad thickening method)
TABLE 4 Effect of different test samples on delayed allergy (x. + -. SD)
Note: * Has significant difference compared with a blank control group
As can be seen from Table 4, after the oral administration of the different test samples 31d to the mice, the swelling degree of the foot sole of the mice in each formulation group is compared with that of the blank control group, the swelling degree of the foot sole of the mice in the example 1 group is larger than that in the control group 2, and the swelling degree of the foot sole of the mice in the example 1 group is significantly different from that in the control group 2 (P < 0.05).
2.3 different oligosaccharide formulations the effect of the test samples on the carbon clearance ability of mice
TABLE 5 different oligosaccharide formulations given the Effect of test samples on carbon clearance Capacity (x + -SD)
| Group of | Animal number (only) | Phagocytic index (a) | P value |
| Blank control group | 12 | 5.01±1.20 | —— |
| EXAMPLE 1 group | 12 | 6.39±0.88 ** | 0.004 |
| Comparative example 1 group | 12 | 5.86±0.98 | 0.072 |
| Comparative example 2 group | 12 | 6.15±0.87 * | 0.015 |
Note: * Significant differences compared with the blank control group; * Very significant differences compared to the blank control group
As can be seen from Table 5, after the mice were orally administered with the different test samples 31d, the mice of the group of example 1 showed an increase in phagocytosis index with a very significant difference (P < 0.01) compared with the blank control group, and the mice of the group of comparative example 2 showed an increase in phagocytosis index with a significant difference (P < 0.05).
3. Small knot
After 31 days of oral administration of different test samples to the mice, compared with the samples of the blank control group and the comparative example 2, the group of the example 1 can obviously increase the toe swelling degree (P is less than 0.05) of the mice and can improve the carbon clearance capacity (P is less than 0.01) of the mice, which indicates that the group of the example 1 can obviously improve the cellular immunity and the non-specific immunity of the mice.
Example 2-example 4:
the weight of each raw material in examples 2 to 4 is shown in table 6 below.
Table 6 weight of each raw material in examples 2 to 4
| Formulation components | |
| Example 2 | 100g of collagen peptide powder, 35g of chitosan oligosaccharide powder, 45g of donkey-hide gelatin powder, 2g of abelmoschus manihot powder and 18g of honey powder |
| Example 3 | 120g of collagen peptide powder, 30g of chitosan oligosaccharide powder, 30g of donkey-hide gelatin powder, 2g of abelmoschus manihot powder and 18g of honey powder |
| Example 4 | 100g of collagen peptide powder, 45g of chitosan oligosaccharide powder, 35g of donkey-hide gelatin powder, 2g of abelmoschus manihot powder and 18g of honey powder |
The practical application condition is as follows:
a total of 200 elderly people 35-70 years old in 5 communities were selected and randomly divided into groups A, B, C, D, E, each of which had 40 people. A. B, C and D take the composition of the powdered chitosan oligosaccharide compound donkey-hide gelatin protein peptide provided by the embodiments 1-4 of the invention respectively, and E takes the formula of the comparative example 1 and are brewed with warm water; the composition is administered once daily in the morning for 3 months.
A. The groups B, C and D all reflect no cold symptoms during the period of taking the composition, and have ruddy complexion, good complexion and enhanced vitality. The product of the invention can enhance the immunity of human body, improve the sub-health state and keep away from the occurrence of diseases.
And the group E exhibited symptoms of body flaming such as redness of eyes, ulceration of the corners of the mouth, swelling of lips, itching of the skin during the first month of administration, and disappeared after the second and third months of administration of the composition of example 1 of the present invention instead.
The above-described embodiments should not be construed as limiting the scope of the present invention, and any alternative modifications or alterations to the embodiments of the present invention will be apparent to those skilled in the art.
The present invention is not described in detail, but is known to those skilled in the art.
Claims (5)
1. The composition of the chitosan oligosaccharide and donkey-hide gelatin protein peptide is characterized in that active ingredients of the composition are prepared from the following raw materials in parts by weight: 10-30 parts of collagen peptide, 2.5-10 parts of chitosan oligosaccharide, 2.5-10 parts of donkey-hide gelatin, 0.05-0.25 part of abelmoschus manihot and 0.5-5 parts of honey powder.
2. The composition of chitosan oligosaccharide compounded donkey-hide gelatin protein peptide as claimed in claim 1, wherein the collagen protein peptide is in the form of granule or powder, the particle size of the granular collagen protein peptide is 20-30 mesh, the particle size of the powdered collagen protein peptide is 120-150 mesh, the particle size of chitosan oligosaccharide is 120-140 mesh, the particle size of donkey-hide gelatin is 120-150 mesh, the particle size of abelmoschus manihot is 120-140 mesh, and the particle size of honey powder is 120-140 mesh.
3. The composition of the chitosan oligosaccharide complex colla corii asini protein peptide according to claim 1, wherein the dosage form of the composition is any one of powder, granules and solution.
4. The method for preparing a composition of chitosan oligosaccharide and donkey-hide gelatin protein peptide according to claim 1, comprising the following steps:
adding the raw materials for forming the active ingredients into a mixer, uniformly mixing, sterilizing and subpackaging to obtain the chitosan oligosaccharide compound donkey-hide gelatin protein peptide composition powder.
5. The preparation method of the composition of chitosan oligosaccharide compound donkey-hide gelatin protein peptide as claimed in claim 1, characterized in that the raw materials for forming the active ingredients are added into a mixer to be mixed uniformly, dissolved into pure water while stirring, then added with active carbon to clarify and remove impurities, and finally filtered, sterilized and filled to obtain the solution of the composition of chitosan oligosaccharide compound donkey-hide gelatin protein peptide.
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