Antarctic krill oil chondroitin extract, antarctic krill oil chondroitin capsule and preparation method

Abstract

The application discloses a euphausia superba oil cartilage extract, a euphausia superba oil cartilage capsule and a preparation method. The euphausia superba oil cartilage extract is composed of the following raw materials in percentage by weight: cartilage collagen 40-60%; non-denatured type II collagen 0-5%; chicken cartilage extract 1-10%; chitosan oligosaccharide 0-10%; and the rest is euphausia superba oil. The compound formula design combines the euphausia superba oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide and other beneficial ingredients, which can produce positive effects on human cardiovascular health, joint health, skin health, immune system health and intestinal health and other aspects, meet the needs of human body in different physiological and health demands, help to improve the quality of life and promote health.

Description

Technical Field

[0001] This invention relates to an edible oil product, and more particularly to an Antarctic krill oil cartilage extract, Antarctic krill oil cartilage capsules, and a preparation method thereof. Background Technology

[0002] The global health and nutrition supplement market is currently experiencing unprecedented growth. Consumer demand for products that improve heart health, enhance brain function, and protect vision is increasing, especially for supplements containing Omega-3 polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Furthermore, with increasing emphasis on beauty, joint health, and gut health, supplements rich in chondroitin, chitosan oligosaccharides, and chicken cartilage extract are also gaining popularity.

[0003] However, existing supplements do not fully meet market demand. Firstly, many supplements containing Omega-3 fatty acids, especially fish oil products, are not efficiently absorbed and utilized by the human body, primarily because the Omega-3 fatty acids exist in the form of glycerides. Secondly, some products on the market may contain heavy metal contamination and other environmental pollutants, posing potential threats to human health. Furthermore, overfishing of marine resources and environmental pollution have made sustainability a key concern.

[0004] In terms of cartilage health, cartilage collagen, chicken cartilage extract, and other ingredients have been proven to improve joint health and skin elasticity. However, they are usually found alone in existing supplements and cannot be effectively combined with other ingredients such as Omega-3 fatty acids to enhance the overall effect of the product.

[0005] Regarding gut health, although oligosaccharides such as chitosan oligosaccharides have been shown to promote gut health, improve the body's absorption of nutrients, and help strengthen the immune system, current products do not effectively combine them with other ingredients to enhance overall health benefits.

[0006] Based on the above, there is an urgent need in the market for a new type of health supplement that can overcome the shortcomings of existing products and provide comprehensive health benefits in a highly effective manner. This supplement should contain easily absorbed Omega-3 fatty acids, while also incorporating beneficial ingredients such as chondroitin collagen, chicken cartilage extract, and chitosan oligosaccharides. These ingredients should work synergistically to enhance their effects on heart health, brain function, vision protection, joint health, skin elasticity, and gut health. Furthermore, this supplement should also consider environmental and sustainability considerations, avoiding further pressure on the environment and marine resources.

[0007] In summary, developing such a novel health supplement has the potential to meet market demand, improve people's quality of life, and potentially open up new avenues for the health and nutritional supplement market. However, this requires researchers to conduct extensive studies and experiments in areas such as raw material selection, formula design, production process development, and product efficacy verification. Summary of the Invention

[0008] To address the shortcomings of existing technologies, the technical problem to be solved by this invention is to provide an Antarctic krill oil cartilage extract, Antarctic krill oil cartilage capsules, and a preparation method thereof.

[0009] The detailed technical solution of this invention is as follows:

[0010] This invention provides an Antarctic krill oil cartilage extract, which is composed of Antarctic krill oil, cartilage collagen, non-denatured type II collagen, and chicken cartilage extract.

[0011] As a preferred embodiment, the Antarctic krill oil cartilage extract also includes chitosan oligosaccharides.

[0012] As a preferred embodiment, the Antarctic krill oil cartilage extract is composed of the following raw materials in weight percentages:

[0013] Cartilage collagen 40-60%;

[0014] 0-5% non-denatured type II collagen;

[0015] Chicken cartilage extract 1-10%;

[0016] Chitosan oligosaccharides 0-10%;

[0017] The remainder is Antarctic krill oil.

[0018] As a preferred embodiment, the Antarctic krill oil cartilage extract is composed of the following raw materials in weight percentages:

[0019] Cartilage collagen 45-55%;

[0020] 0.2-2% non-denatured type II collagen;

[0021] Chicken cartilage extract 2-8%;

[0022] Chitosan oligosaccharides 0.2-5%;

[0023] The remainder is Antarctic krill oil.

[0024] As a preferred embodiment, the Antarctic krill oil cartilage extract is composed of the following raw materials in weight percentages:

[0025] Cartilage collagen 47-51%;

[0026] Non-denatured type II collagen 0.5-1.5%;

[0027] Chicken cartilage extract 3-7%;

[0028] Chitosan oligosaccharides 1-3%;

[0029] The remainder is Antarctic krill oil.

[0030] This invention also provides a method for preparing Antarctic krill oil, comprising the following steps:

[0031] 1. Pulverization: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0032] 2. Extraction: Antarctic krill powder was extracted using organic solvents;

[0033] 3. Separation: The organic solvent and Antarctic krill powder are separated by filtration;

[0034] 4. Vacuum distillation.

[0035] As a preferred method, an extraction step for preparing Antarctic krill oil is as follows: Antarctic krill powder is mixed with 2-6 times its mass of organic solvent, and then stirred at 50-100 rpm for 1-3 hours at a temperature of 20-30°C; then the mixture is allowed to stand for 1-3 hours at the same temperature.

[0036] As a preferred embodiment, a method for preparing Antarctic krill oil includes the following steps:

[0037] 1. Pulverization: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0038] 2. Enzymatic hydrolysis: Add enzymes to Antarctic krill powder for enzymatic hydrolysis;

[0039] 3. Extraction: Extract the enzymatically hydrolyzed Antarctic krill powder using an organic solvent;

[0040] 4. Separation: The organic solvent and Antarctic krill powder are separated by filtration;

[0041] 5. Vacuum distillation.

[0042] As a preferred embodiment, a method for preparing Antarctic krill oil includes the following steps:

[0043] Pulverizing step: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0044] Enzymatic hydrolysis step: Add 1-3 times the amount of water and enzyme to Antarctic krill powder, maintain at 20-40℃, and stir at 30-80 rpm for 2-4 hours. The amount of enzyme used is 1000-50000 units per gram of Antarctic krill powder.

[0045] Extraction steps: Mix the enzymatically hydrolyzed Antarctic krill powder with 2-6 times its weight of organic solvent, and then stir at 50-100 rpm for 1-3 hours at 20-30°C; then let the mixture stand for 1-3 hours at the same temperature; preferably, the organic solvent is ethanol.

[0046] Separation steps: The mixture is filtered and separated, and then rotary evaporated to obtain an oil-containing ethanol solution;

[0047] Vacuum distillation procedure: Distill the oil-containing ethanol solution under reduced pressure.

[0048] Preferably, the enzyme is at least one of cholesterol esterase and protease.

[0049] As a preferred embodiment, the Antarctic krill oil cartilage extract is prepared using the method described above.

[0050] The present invention also provides a method for preparing Antarctic krill oil cartilage extract, comprising the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide evenly.

[0051] The present invention also provides an Antarctic krill oil cartilage capsule, which consists of the aforementioned Antarctic krill oil cartilage extract and a soft capsule encapsulating the Antarctic krill oil cartilage extract.

[0052] This invention also provides a method for preparing Antarctic krill oil cartilage capsules, comprising the following steps:

[0053] Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract, and chitosan oligosaccharide were mixed to obtain Antarctic krill oil cartilage extract.

[0054] Antarctic krill oil cartilage extract is pressed into soft capsules;

[0055] Finalization;

[0056] drying;

[0057] Filling.

[0058] This invention also provides the application of Antarctic krill oil cartilage extract in the preparation of Antarctic krill oil cartilage capsules.

[0059] This invention features a compound formulation design that combines various beneficial ingredients such as Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract, and chitosan oligosaccharides. This formulation may have positive effects on multiple aspects of human health, including cardiovascular health, joint health, skin health, immune system health, and intestinal health. It meets the body's needs in various physiological and health aspects, helps improve quality of life, and promotes health. Detailed Implementation

[0060] An Antarctic krill oil cartilage extract, comprising the following ingredients by weight percentage:

[0061] Cartilage collagen 47-51%;

[0062] Non-denatured type II collagen 0.5-1.5%;

[0063] Chicken cartilage extract 3-7%;

[0064] Chitosan oligosaccharides 1-3%;

[0065] The remainder is Antarctic krill oil.

[0066] Antarctic krill oil, a highly nutritious health supplement rich in Omega-3 polyunsaturated fatty acids, has been scientifically proven to have significant effects on maintaining heart health, improving brain function, and protecting vision. The Omega-3 fatty acids in this supplement, primarily eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), can exert various physiological functions in the body, including anti-inflammatory, anticoagulant, and immune-regulating effects, thereby reducing the risk of heart disease and stroke. The importance of DHA in the brain is also undeniable; it helps maintain the integrity of nerve cell membranes, promotes the release of neurotransmitters, regulates neuroinflammatory responses, thereby improving memory and learning abilities and reducing the risk of cognitive impairment. Regarding vision protection, as a key component of retinal photoreceptor cells, adequate DHA intake helps prevent vision degeneration and maintain good vision. Furthermore, the Omega-3 fatty acids in Antarctic krill oil exist in the form of phospholipids, which have higher bioavailability than the glycerides in fish oil supplements, making them easier for the body to absorb and further enhancing the health benefits of Antarctic krill oil.

[0067] Collagen is the most abundant protein in the human body, accounting for nearly one-third of total protein, with the main types being types I, II, and III. Cartilage collagen is primarily type II collagen, mainly found in human cartilage, and plays a crucial role in maintaining joint health. Cartilage collagen helps maintain joint elasticity and flexibility by providing structural support to cartilage, and prevents cartilage wear and degeneration. Additionally, cartilage collagen also functions in the skin, helping to maintain its elasticity and firmness.

[0068] Undenatured type II collagen is a special form of collagen that retains the original triple helix structure. This type of collagen can exert a therapeutic effect on rheumatoid arthritis and other autoimmune arthritis through an oral immune tolerance mechanism. This mechanism involves the regulation of T cells in the immune system. The antigenic determinants on undenatured type II collagen can form a lock-key mechanism with T cells generated from Peyresia aggregates in the gut, inhibiting the activity of killer T cells, thereby controlling the immune and inflammatory responses, further halting the progression of arthritis, and allowing joints to heal and regenerate.

[0069] Chicken cartilage extract is a substance extracted from chicken cartilage, containing a high amount of chondroitin sulfate. Chondroitin sulfate is a large-molecule polysaccharide found in cartilage, tendons, ligaments, and skin. Chondroitin sulfate plays an important role in maintaining joint health, helping to absorb impact and maintain cartilage elasticity. Furthermore, chondroitin sulfate has anti-inflammatory properties, inhibiting the production and release of inflammatory cytokines, thereby alleviating symptoms of inflammatory diseases such as arthritis.

[0070] Chitosan oligosaccharides are oligosaccharides composed of 2-10 monosaccharide units, primarily derived from shrimp and crab shells. Considered a prebiotic, chitosan oligosaccharides can promote the growth of beneficial gut microbiota and inhibit the growth of harmful bacteria, thus offering significant benefits to gut health. They can enhance the function of the intestinal mucosal barrier, reducing the invasion of harmful substances, and stimulate the intestinal immune system, thereby improving immunity. Furthermore, chitosan oligosaccharides help regulate gut microbiota balance, improve constipation, and lower blood lipids and blood sugar, offering potential benefits for patients with diabetes and cardiovascular disease.

[0071] This invention also provides a method for preparing Antarctic krill oil, comprising the following steps:

[0072] 1. Pulverization: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0073] 2. Extraction: Antarctic krill powder was extracted using organic solvents;

[0074] 3. Separation: The organic solvent and Antarctic krill powder are separated by filtration;

[0075] 4. Vacuum distillation.

[0076] As a preferred method, an extraction step for preparing Antarctic krill oil is as follows: Antarctic krill powder is mixed with 2-6 times its mass of organic solvent, and then stirred at 50-100 rpm for 1-3 hours at a temperature of 20-30°C; then the mixture is allowed to stand for 1-3 hours at the same temperature.

[0077] As a preferred embodiment, a method for preparing Antarctic krill oil includes the following steps:

[0078] 1. Pulverization: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0079] 2. Enzymatic hydrolysis: Add enzymes to Antarctic krill powder for enzymatic hydrolysis;

[0080] 3. Extraction: Extract the enzymatically hydrolyzed Antarctic krill powder using an organic solvent;

[0081] 4. Separation: The organic solvent and Antarctic krill powder are separated by filtration;

[0082] 5. Vacuum distillation.

[0083] As a preferred embodiment, a method for preparing Antarctic krill oil includes the following steps:

[0084] Pulverizing step: Antarctic krill is freeze-dried, pulverized, and ground to obtain Antarctic krill powder;

[0085] Enzymatic hydrolysis step: Add 1-3 times the amount of water and enzyme to Antarctic krill powder, maintain at 20-40℃, and stir at 30-80 rpm for 2-4 hours. The amount of enzyme used is 1000-50000 units per gram of Antarctic krill powder.

[0086] Extraction steps: Mix the enzymatically hydrolyzed Antarctic krill powder with 2-6 times its weight of organic solvent, and then stir at 50-100 rpm for 1-3 hours at 20-30°C; then let the mixture stand for 1-3 hours at the same temperature; preferably, the organic solvent is ethanol.

[0087] Separation steps: The mixture is filtered and separated, and then rotary evaporated to obtain an oil-containing ethanol solution;

[0088] Vacuum distillation procedure: Distill the oil-containing ethanol solution under reduced pressure.

[0089] Preferably, the enzyme is at least one of cholesterol esterase and protease. More preferably, the enzyme is composed of cholesterol esterase and protease in a mass ratio of (1-3):(1-3).

[0090] Cholesterol esterase is a catalytic enzyme that plays a crucial role in the preparation of Antarctic krill oil. It esterifies free cholesterol and fatty acids in the krill powder to form cholesterol esters. This process is an esterification reaction, where ester bonds are formed between cholesterol and fatty acids, tightly binding them together. Cholesterol esters are the product of the combination of fatty acids and cholesterol, and they constitute a significant proportion of Antarctic krill oil.

[0091] The inventors discovered that cholesterol esterase and protease can effectively improve performance, significantly increasing the astaxanthin extraction rate from Antarctic krill. The possible mechanistic explanation for why cholesterol esterase and protease can effectively improve the astaxanthin extraction rate from Antarctic krill is as follows:

[0092] (1) Action of cholesterol esterase: Cholesterol esterase is an enzyme that promotes the hydrolysis of cholesterol esters, breaking them down into free cholesterol and fatty acids. In Antarctic krill oil, a certain amount of cholesterol esters may be bound to astaxanthin, making it difficult for astaxanthin to be fully released. By adding cholesterol esterase, the hydrolysis process of cholesterol esters can be accelerated, allowing astaxanthin to be released from the cholesterol esters and improving the extractability of astaxanthin.

[0093] (2) Action of proteases: Proteases are a class of enzymes that catalyze the hydrolysis of proteins, breaking them down into amino acids or small peptides. In Antarctic krill, astaxanthin may bind to proteins to form complexes, making it difficult to extract astaxanthin from these proteins. By adding proteases, proteins can be hydrolyzed into amino acids or small peptides, disrupting the binding of astaxanthin to proteins and thus increasing the extractability of astaxanthin.

[0094] In summary, cholesterol esterase and protease can respectively solve the problem of astaxanthin binding to cholesterol esters and proteins, thereby improving the extraction rate of astaxanthin from Antarctic krill. The addition of these enzymes can effectively release and dissociate astaxanthin, making it easier to extract astaxanthin from Antarctic krill, thus increasing the effective extraction rate of astaxanthin.

[0095] Obviously, not all proteases are suitable. The inventors further screened the proteases through numerous experiments, ultimately obtaining at least one of clostridial protease, alkaline protease, papain, and neutral protease. Preferably, the protease is clostridial protease. Most preferably, the enzyme is composed of cholesterol esterase and clostridial protease in a mass ratio of (1-3):(1-3).

[0096] Clostridium protease is a proteolytic enzyme isolated from Clostridium histolyticum, possessing esterase, amidase, and protease activities. Alkaline protease is a proteolytic enzyme. Papain is a cysteine ​​protease of the peptidase C1 family, used in the food, pharmaceutical, textile, and cosmetic industries. Neutral protease is a neutral protease derived from microorganisms, used for cell and tissue isolation.

[0097] Clostridium protease, a serine protease, may have a molecular structure better suited for specific interactions with astaxanthin complexes. This unique structure may give it better affinity and specificity in the astaxanthin-bound state, thus facilitating the release of astaxanthin from the complex. As a serine enzyme, clostridium protease exhibits high specificity, primarily catalyzing the hydrolysis of serine residues. When astaxanthin binds to serine residues in proteins to form complexes, clostridium protease may more readily disrupt these complexes, facilitating the release of astaxanthin. This serine enzyme specificity may enable clostridium protease to more effectively recognize astaxanthin complexes and promote the release of astaxanthin from the complex during enzymatic hydrolysis. Therefore, clostridium protease more effectively improves the extraction rate of astaxanthin from Antarctic krill compared to other types of proteases.

[0098] As a preferred embodiment, the Antarctic krill oil cartilage extract is prepared using the method described above.

[0099] The present invention also provides a method for preparing Antarctic krill oil cartilage extract, comprising the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide evenly.

[0100] The present invention also provides an Antarctic krill oil cartilage capsule, which consists of the aforementioned Antarctic krill oil cartilage extract and a soft capsule encapsulating the Antarctic krill oil cartilage extract.

[0101] This invention also provides a method for preparing Antarctic krill oil cartilage capsules, comprising the following steps:

[0102] Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract, and chitosan oligosaccharide were mixed to obtain Antarctic krill oil cartilage extract.

[0103] Antarctic krill oil cartilage extract is pressed into soft capsules;

[0104] Finalization;

[0105] drying;

[0106] Filling.

[0107] As a preferred embodiment, a method for preparing Antarctic krill oil cartilage capsules includes the following steps:

[0108] Add the appropriate parameters of Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract, and chitosan oligosaccharide to the mixing tank in advance as required.

[0109] Start the mixer and mix for 12-24 hours. Take samples at different times for testing. When the test results are stable, the mixture is considered to be homogeneous and the Antarctic krill oil cartilage extract is obtained.

[0110] Mix gelatin, glycerin, and water in a mass ratio of 1:(0.2-0.6):(0.6-1) until homogeneous, heat to 60-90℃, and maintain for 1-3 hours; if necessary, you can also add appropriate amounts of natural carotene and beetroot red.

[0111] Once there are no gelatin particles to be detected, begin vacuuming at a negative pressure of 0.04-0.06 MPa for 0.5-2 hours.

[0112] Antarctic krill oil cartilage extract is pressed into soft capsules using soft capsule production equipment;

[0113] Setting: Temperature 20-40℃, humidity 15-40%, time 0.5-2 hours;

[0114] Drying: Temperature 20-40℃, humidity 15-40%, time 12-24 hours;

[0115] Filling process: control the amount of soft capsules in each capsule to be 0.2-0.3 grams and the amount of Antarctic krill oil cartilage extract to be 0.9-1.1 grams.

[0116] The present invention also provides the application of Antarctic krill oil cartilage extract in the preparation of Antarctic krill oil cartilage capsules.

[0117] Antarctic krill oil cartilage extract has several positive and beneficial effects:

[0118] (1) Maintaining heart and brain health: Omega-3 fatty acids in Antarctic krill oil, especially EPA and DHA, help reduce the risk of heart disease and stroke, while improving brain function and helping to maintain healthy cognitive abilities and vision.

[0119] (2) Protecting joints: The presence of cartilage collagen and non-denatured type II collagen helps maintain joint health, promotes cartilage elasticity and prevents cartilage degeneration, while inhibiting the development of arthritis.

[0120] (3) Improve skin condition: Supplementing with cartilage collagen helps maintain skin elasticity and firmness, and promotes healthy skin.

[0121] (4) Boost immunity: The effects of non-denatured type II collagen and chitosan oligosaccharide help enhance the function of the immune system, improve the body's resistance, and make the body more capable of resisting diseases.

[0122] (5) Promote gut health: Chitosan oligosaccharide helps maintain the balance of gut microbiota and promotes the growth of beneficial bacteria, thereby improving digestive function and alleviating intestinal problems such as constipation.

[0123] Overall, the Antarctic krill oil cartilage extract compound formulation of this invention has beneficial effects in multiple aspects, covering important areas such as cardiovascular health, joint health, skin health, immune system health, and gut health. However, it is worth noting that individuals should carefully select supplements based on their own health conditions and needs, and follow the advice of a doctor or healthcare professional.

[0124] In the following examples:

[0125] Hydrolyzed type II collagen, also known as chondroitin collagen, is sold by Anhui Shengmeinuo Biotechnology Co., Ltd., and conforms to the type A standard in Q / ASMN 0010S-2020.

[0126] The non-denatured type II collagen was selected from the non-denatured cartilage collagen sold by Anhui Shengmeinuo Biotechnology Co., Ltd., which was prepared using ZL2018111757422.

[0127] Chicken cartilage extract, specifically the chicken cartilage extract from Anhui Shengmeinuo Biotechnology Co., Ltd. (product number ZHOJ22002938.001), with a chicken cartilage extract content of 31.1 wt%. Standard implemented: Q / ASMN 0009S-2020.

[0128] Chitosan oligosaccharide: 2-Acetamido-2-deoxy-D-glucose, CAS No.: 7512-17-6, sourced from Amexco Co., Ltd., South Korea. N-acetylglucosamine (NAG).

[0129] Cholesterol esterase, CAS No.: 9026-00-0, was selected from 10KU cholesterol esterase provided by Hefei Bomei Biotechnology Co., Ltd.

[0130] Clostridium protease, CAS No.: 9028-00-6, provided by Shanghai Baoman Biotechnology Co., Ltd., selected as 10KU.

[0131] Alkaline protease, CAS No.: 9014-01-1, provided by Shanghai Jianglai Biotechnology Co., Ltd., selected as 10KU. It is a proteolytic enzyme obtained by deep fermentation, extraction, and purification of *Bacillus licheniformis* strain 2709, cultivated through fermentation or bacterial protoplast mutagenesis. It belongs to the serine-bound, highly alkaline protease family. Its main component is *Bacillus subtilis* protease, an endonuclease catalyzing serine.

[0132] Papain, CAS No. 9001-73-4, provided by Guangdong Manxin Biotechnology Co., Ltd., selected as 10KU. Papain is a thiol protease with broad substrate specificity, acting on peptide bonds formed by the carboxyl groups of L-arginine, L-lysine, glycine, and L-papain amino acid residues in proteins. This enzyme is an endopeptidase, capable of cleaving the internal peptide chain —CO—NH— of whole egg proteins to generate smaller polypeptides.

[0133] Type A Antarctic krill oil is prepared using the following method:

[0134] Pulverizing step: Freeze-dry Antarctic krill, pulverize and grind it, and pass it through a 50-mesh sieve to obtain Antarctic krill powder;

[0135] Extraction steps: Mix Antarctic krill powder with 4 times its weight of organic solvent ethanol, and then stir at 80 rpm for 2 hours at 25°C; then let the mixture stand for 2 hours at the same temperature.

[0136] Separation steps: The mixture is separated by filtration through a 0.2 mm pore size filter, followed by rotary evaporation (heating temperature 50℃, rotary evaporation speed set at 150 rpm) to obtain an oil-containing ethanol solution;

[0137] Vacuum distillation procedure: The oil-containing ethanol solution is distilled under reduced pressure at a temperature of 75°C and a pressure of 30 mmHg to obtain type A Antarctic krill oil.

[0138] The preparation method is basically the same as that of another product of the inventor, Haematococcus pluvialis astaxanthin oil.

[0139] Type B Antarctic krill oil is prepared using the following method:

[0140] Pulverizing step: Freeze-dry Antarctic krill, pulverize and grind it, and pass it through a 50-mesh sieve to obtain Antarctic krill powder;

[0141] Enzymatic hydrolysis steps: Add twice the amount of water and enzyme to Antarctic krill powder, maintain at 30°C, and stir at 50 rpm for 3 hours. The amount of enzyme used is 10,000 units per gram of Antarctic krill powder.

[0142] Extraction steps: Mix the enzymatically hydrolyzed Antarctic krill powder with 4 times its mass of organic solvent ethanol, and then stir at 80 rpm for 2 hours at 25°C; then let the mixture stand for 2 hours at the same temperature.

[0143] Separation steps: The mixture is separated by filtration through a 0.2 mm pore size filter, followed by rotary evaporation (heating temperature 50℃, rotary evaporation speed set at 150 rpm) to obtain an oil-containing ethanol solution;

[0144] Vacuum distillation procedure: The oil-containing ethanol solution is distilled under reduced pressure at a temperature of 75°C and a pressure of 30 mmHg to obtain type B Antarctic krill oil.

[0145] The enzyme used in the enzymatic hydrolysis step is cholesterol esterase.

[0146] Type C Antarctic krill oil is prepared using the following method:

[0147] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is clostridium protease.

[0148] Type D Antarctic krill oil is prepared using the following method:

[0149] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is a mixture of cholesterol esterase and clostridium protease in a mass ratio of 3:1.

[0150] Type E Antarctic krill oil is prepared using the following method:

[0151] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is a mixture of cholesterol esterase and clostridium protease in a mass ratio of 1:3.

[0152] Type F Antarctic krill oil is prepared using the following method:

[0153] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is a mixture of cholesterol esterase and clostridium protease in a 1:1 mass ratio.

[0154] Type G Antarctic krill oil is prepared using the following method:

[0155] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is alkaline protease.

[0156] H-type Antarctic krill oil is prepared using the following method:

[0157] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is papain.

[0158] Type I Antarctic krill oil is prepared using the following method:

[0159] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is a mixture of cholesterol esterase and alkaline protease in a 1:1 mass ratio.

[0160] J-type Antarctic krill oil is prepared using the following method:

[0161] The preparation process is basically the same as that of type B Antarctic krill oil, except that the enzyme used in the enzymatic hydrolysis step is a mixture of cholesterol esterase and papain in a 1:1 mass ratio.

[0162] Example 1

[0163] Antarctic krill oil cartilage extract, composed of the following ingredients by weight percentage:

[0164] Cartilage collagen 49%;

[0165] 1% non-denatured type II collagen;

[0166] 5% chicken cartilage extract;

[0167] Chitosan oligosaccharide 2%;

[0168] Type A Antarctic krill oil 43%.

[0169] The preparation method of Antarctic krill oil cartilage extract includes the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide evenly.

[0170] Comparative Example 1

[0171] Antarctic krill oil cartilage extract, composed of the following ingredients by weight percentage:

[0172] Cartilage collagen 49%;

[0173] 5% chicken cartilage extract;

[0174] Chitosan oligosaccharide 2%;

[0175] Type A Antarctic krill oil 44%.

[0176] The preparation method of Antarctic krill oil cartilage extract includes the following steps: mixing Antarctic krill oil, cartilage collagen, chicken cartilage extract and chitosan oligosaccharide evenly.

[0177] Comparative Example 2

[0178] Antarctic krill oil cartilage extract, composed of the following ingredients by weight percentage:

[0179] Cartilage collagen 49%;

[0180] 1% undenatured type II collagen;

[0181] Chitosan oligosaccharide 2%;

[0182] Type A Antarctic krill oil 48%.

[0183] The preparation method of the Antarctic krill oil cartilage extract includes the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen and chitosan oligosaccharide evenly.

[0184] Comparative Example 3

[0185] Antarctic krill oil cartilage extract, composed of the following ingredients by weight percentage:

[0186] Cartilage collagen 49%;

[0187] 1% undenatured type II collagen;

[0188] 5% chicken cartilage extract;

[0189] Type A Antarctic krill oil 45%.

[0190] The preparation method of Antarctic krill oil cartilage extract includes the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen and chicken cartilage extract evenly.

[0191] Example 2

[0192] Antarctic krill oil cartilage extract, composed of the following ingredients by weight percentage:

[0193] Cartilage collagen 49%;

[0194] 1% undenatured type II collagen;

[0195] 5% chicken cartilage extract;

[0196] Chitosan oligosaccharide 2%;

[0197] Type F Antarctic krill oil 43%.

[0198] The preparation method of Antarctic krill oil cartilage extract includes the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide evenly.

[0199] Test Example 1: Test for Improving Osteoporosis

[0200] Number of participants in each group: 12 patients with osteoporosis.

[0201] Oral dosage: The experimental group took 2 grams of Antarctic krill oil cartilage extract daily, while the control group took 2 grams of placebo.

[0202] Experiment duration: 70 days.

[0203] The average value of the test results is taken.

[0204] Table 1: Osteocalcin Enhancement Rate

[0205]

[0206] The data in the table above shows that the use of cholesterol esterase and protease can effectively improve performance, significantly increasing the astaxanthin extraction rate from Antarctic krill. The possible mechanistic reasons why cholesterol esterase and protease can effectively improve the astaxanthin extraction rate from Antarctic krill are as follows:

[0207] (1) Action of cholesterol esterase: Cholesterol esterase is an enzyme that promotes the hydrolysis of cholesterol esters, breaking them down into free cholesterol and fatty acids. In Antarctic krill oil, a certain amount of cholesterol esters may be bound to astaxanthin, making it difficult for astaxanthin to be fully released. By adding cholesterol esterase, the hydrolysis process of cholesterol esters can be accelerated, allowing astaxanthin to be released from the cholesterol esters and improving the extractability of astaxanthin.

[0208] (2) Action of proteases: Proteases are a class of enzymes that catalyze the hydrolysis of proteins, breaking them down into amino acids or small peptides. In Antarctic krill, astaxanthin may bind to proteins to form complexes, making it difficult to extract astaxanthin from these proteins. By adding proteases, proteins can be hydrolyzed into amino acids or small peptides, disrupting the binding of astaxanthin to proteins and thus increasing the extractability of astaxanthin.

[0209] In summary, cholesterol esterase and protease can respectively solve the problem of astaxanthin binding to cholesterol esters and proteins, thereby improving the extraction rate of astaxanthin from Antarctic krill. The addition of these enzymes can effectively release and dissociate astaxanthin, making it easier to extract astaxanthin from Antarctic krill, thus increasing the effective extraction rate of astaxanthin.

[0210] Test Example 2:

[0211] Determination of astaxanthin content in Antarctic krill meal:

[0212] Weigh approximately 25 mg of pretreated Antarctic krill powder and place it in a 10 ml centrifuge tube. Add 3 g of quartz sand and 5 ml of DMSO to the centrifuge tube. Incubate in a 50°C water bath for 15 min, shaking three times during the incubation period. Centrifuge at 4000 rpm for 3 min to precipitate the insoluble matter. Transfer the supernatant to a 25 ml volumetric flask. Add 5 ml of n-hexane to the centrifuge tube, vortex for 30 s, centrifuge to separate the precipitate, and transfer the supernatant to a 25 ml volumetric flask. Repeat the n-hexane extraction until the supernatant is essentially colorless (absorbance less than 0.05). Dilute to volume with n-hexane as needed, and measure the absorbance at 474 nm using n-hexane as a blank (between 0.25 and 0.75). Calculate using the following formula:

[0213] Astaxanthin content (%) in Antarctic krill meal = A × 25 × dilution factor ÷ [2100 × mass of Antarctic krill meal (g)] × 80%

[0214] In the formula: 2100 is the absorbance coefficient of a 1% (m / v) astaxanthin n-hexane standard solution at 474 nm.

[0215] Determination of astaxanthin content in Antarctic krill oil:

[0216] Weigh approximately 25 mg of Antarctic krill oil into a 100 ml volumetric flask, dissolve and dilute to volume with n-hexane, dilute appropriately, and measure the absorbance at 474 nm (between 0.25 and 0.75) using n-hexane as a blank. Calculate using the following formula:

[0217] Astaxanthin content (%) in Antarctic krill oil = A × 25 × dilution factor ÷ [2100 × mass of Antarctic krill oil (g)] × 80%

[0218] In the formula: 2100 is the absorbance coefficient of a 1% (m / v) astaxanthin n-hexane standard solution at 474 nm.

[0219] The extraction rate refers to the astaxanthin extraction rate from Antarctic krill powder, calculated using the following formula:

[0220] Astaxanthin extraction rate (%) = Mass of Antarctic krill oil × Astaxanthin content in Antarctic krill oil ÷ Mass of Antarctic krill meal ÷ Astaxanthin content in Antarctic krill meal × 100%

[0221] Table 1: Astaxanthin Extraction Rate Test Table from Antarctic Krill

[0222]

[0223]

[0224] Example 3

[0225] A method for preparing Antarctic krill oil cartilage capsules includes the following steps:

[0226] Mix gelatin, glycerin, and water in a mass ratio of 1:0.4:0.8 until homogeneous, heat to 70°C and maintain for 2 hours; once there are no gelatin particles, begin vacuuming at a negative pressure of 0.05 MPa for 1 hour.

[0227] The Antarctic krill oil cartilage extract from Example 2 was pressed into soft capsules using a soft capsule production equipment;

[0228] Setting: Temperature 30℃, humidity 30%, time 1 hour;

[0229] Drying: Temperature 30℃, humidity 25%, time 18 hours;

[0230] Filling process: control the amount of soft capsules in each capsule to 0.25g and the mass of Antarctic krill oil cartilage extract to 1g.

[0231] Test Example 3:

[0232] The performance of the Antarctic krill oil cartilage extract capsules prepared in Example 3 was tested, as follows:

[0233]

[0234]

[0235] Acid value: Tested according to Method II of GB 5009.229-2016.

[0236] Total phospholipids: Tested according to Method I of GB / T 5537-2008.

[0237] Moisture content: Tested according to Method I of GB 5009.3-2016.

[0238] Total bacterial count: tested according to GB 4789.2-2016.

[0239] Coliform bacteria: tested according to GB 4789.3-2016, Method II.

[0240] Mold: Tested according to GB 4789.15-2016, Method I; Staphylococcus aureus: Tested according to GB 4789.10-2016, Method I; Salmonella: Tested according to GB 4789.4-2016; Peroxide value: Tested according to GB 5009.227-2016, Method I.

[0241] Acid value: Tested according to GB 5009.229-2016, Method II, for sorbic acid and its potassium salt (calculated as sorbic acid); Tested according to GB 5009.28-2016, Method I, for benzoic acid and its sodium salt (calculated as benzoic acid); Tested according to GB 5009.28-2016, Method I, for butylated hydroxytoluene (BHT); Tested according to GB 5009.32-2016, Method IV, for butylated hydroxyanisole (BHA); Tested according to GB 5009.32-2016, Method IV, for lead (calculated as Pb); Tested according to GB 5009.12-2017, Method II, for inorganic arsenic (calculated as As); Tested according to GB 5009.11-2014, Part II, Method I, for energy and carbohydrates; Calculated according to GB 28050-2011; Tested according to GB 5009.6-2016, Method I, for fats.

[0242] Protein: Tested according to Method I of GB 5009.5-2016; Sodium: Tested according to Method IV of GB 5009.91-2017.

Claims

1. An extract of Antarctic krill oil cartilage, characterized in that, It is composed of the following raw materials by weight percentage: Cartilage collagen 45-55%; Non-denatured type II collagen 0.2-2%; Chicken cartilage extract 2-8%; Chitosan oligosaccharide 0.2-5%; The rest is Antarctic krill oil. The Antarctic krill oil is prepared by the following method: A crushing step: Antarctic krill is freeze-dried, crushed and ground to obtain Antarctic krill powder; An enzymolysis step: 1-3 times the amount of water and enzymes is added to the Antarctic krill powder, and the mixture is kept at 20-40℃, stirred at 30-80 rpm for 2-4 hours, and the amount of enzymes used is 1000-50000 units per gram of Antarctic krill powder; An extraction step: the enzymolysis-treated Antarctic krill powder is mixed with 2-6 times its mass of organic solvent, and then stirred at 50-100 rpm for 1-3 hours at a temperature of 20-30℃; then the mixture is allowed to stand at the same temperature for 1-3 hours, and the organic solvent is ethanol; A separation step: the mixture is filtered and rotary evaporated to obtain an oil-containing ethanol solution; A reduced pressure distillation step: the oil-containing ethanol solution is distilled under reduced pressure; The enzymes are composed of cholesterol esterase and clostridial protease in a mass ratio of (1-3):(1-3).

2. The Euphausiids oil chondroitin extract according to claim 1, characterized in that, It is composed of the following raw materials by weight percentage: Cartilage collagen 47-51%; Non-denatured type II collagen 0.5-1.5%; Chicken cartilage extract 3-7%; Chitosan oligosaccharide 1-3%; The rest is Antarctic krill oil.

3. The method for preparing the Euphausia superba oil chondroitin extract according to claim 1 or 2, characterized in that, It comprises the following steps: mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide uniformly.

4. A capsule of Antarctic krill oil chondroitin, characterized by, It is composed of the Antarctic krill oil cartilage extract of claim 1 or 2 and soft capsules containing the Antarctic krill oil cartilage extract.

5. A method of preparing the Euphausia superba oil soft cartilage capsule of claim 4, characterized by, It comprises the following steps: Mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide to obtain Antarctic krill oil cartilage extract; Pressing the Antarctic krill oil cartilage extract into soft capsules; Shaping; Drying; Filling. It comprises the following steps: Mixing Antarctic krill oil, cartilage collagen, non-denatured type II collagen, chicken cartilage extract and chitosan oligosaccharide to obtain Antarctic krill oil cartilage extract; Pressing the Antarctic krill oil cartilage extract into soft capsules; Shaping; Drying; Filling.

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