Protein bar with anti-oxidation and glycolipid metabolism regulation functions and preparation method thereof

Abstract

The invention belongs to the technical field of computer security, and particularly relates to a protein bar with anti-oxidation and glycolipid metabolism regulation functions and a preparation method thereof.The protein bar is of a three-layer structure including an outer layer, a sandwich layer and an inner layer, the sandwich layer serves as a core, functional components formed by compounding penaeus vannamei extracts and Tibetan gastrodia elata water extracts according to the ratio of 3: 7 are adopted, and the protein bar is prepared. Nutrient substances such as whey protein powder and oatmeal are matched as raw materials; the preparation method comprises the following steps: extracting functional components, preparing each layer, treating after forming, performing ultrasonic microwave-assisted enzymolysis, performing water bath reflux extraction to obtain the functional components, and performing low-temperature preparation, hot air baking, low-temperature sterilization and nitrogen-filled packaging to obtain the finished product. Synergistic antioxidation and glycolipid metabolism regulation of the two functional components are achieved, active components are reserved, the fishy smell of the crustacean raw materials is eliminated, high-value utilization of aquatic byproducts and food development of the Tibetan gastrodia elata are further achieved, and the product is comprehensive in nutrition, excellent in taste, safe in quality and suitable for people who are sub-healthy and do exercises and fitness and the like.

Description

Technical Field

[0001] This invention belongs to the field of functional food processing technology, specifically relating to a protein bar with antioxidant and glucose and lipid metabolism regulation functions and its preparation method. Background Technology

[0002] The shrimp heads, shells, and other crustacean byproducts produced during the processing of Litopenaeus vannamei are rich in bioactive peptide precursors. The functional crustacean peptides obtained by enzymatic hydrolysis are easily absorbed, have low allergenicity, and possess activities such as regulating glucose and lipid metabolism and antioxidation. However, their deep processing and integration into functional foods are still in their early stages.

[0003] Tibetan Gastrodia elata from Bomi County, Linzhi City, Tibet, is a geographical indication product. Its water extract contains active ingredients such as gastrodin and polyphenols, which have significant antioxidant activity and can improve cell oxidative damage caused by exogenous toxicity. However, its development is mostly concentrated in the field of traditional Chinese medicine preparations, with insufficient cross-border integration with food.

[0004] Existing protein bars generally suffer from a single function, focusing only on protein supplementation and lacking dual physiological regulatory functions. Furthermore, the raw materials do not incorporate high-value aquatic by-products or authentic Chinese medicinal herbs. Some foods containing crustacean ingredients have a strong fishy smell and their functional components are easily degraded by high temperatures. At the same time, existing formulas are mostly simple combinations of raw materials, failing to explore the synergistic effects of different functional components, making it difficult to meet the diverse health needs of the market.

[0005] To this end, this invention combines the needs of resource recycling and the modernization of traditional Chinese medicine with the scientific compounding of crustacean functional peptides and Tibetan gastrodia elata water extract to develop a protein bar with multiple functions. At the same time, the process is optimized to solve the problems of fishy smell and component degradation, filling the gap in existing products. Summary of the Invention

[0006] To overcome the shortcomings of existing technologies, this product uses shellfish functional peptides prepared by enzymatic hydrolysis of processing by-products from Litopenaeus vannamei and water extracts of Gastrodia elata from Bomi, Linzhi, Tibet as core compound functional ingredients, combined with conventional nutritional raw materials such as whey protein powder and soy protein powder. This achieves dual functions of antioxidation and regulation of glucose and lipid metabolism, while simultaneously supplementing the body with high-quality protein, dietary fiber, vitamins, and other nutrients. Secondly, it provides a standardized preparation method for this protein bar, using low-temperature processing to preserve the activity of functional components, scientifically combining raw materials to eliminate the inherent fishy smell of shellfish, and controlling key points to ensure product quality and safety. Ultimately, it realizes the high-value recycling of aquatic processing by-products and the cross-industry development of Tibetan Gastrodia elata into food products, providing convenient, nutritionally complete, and multifunctional functional foods for fitness enthusiasts, sub-healthy individuals, and general consumers.

[0007] The technical solution adopted by this invention to solve its technical problem is as follows: This invention proposes a protein bar with antioxidant and glucose-lipid metabolism regulation functions. The protein bar has a three-layer structure, including an outer layer, a sandwich layer, and an inner layer. Based on a total mass of 100g for each layer, the components and amounts added to each layer are as follows:

[0008] Each 100g of the outer layer contains 12g of chocolate and 4.8g of chopped nuts;

[0009] Each 100g of the sandwich layer contains 6.5g of whey protein powder, 2.5g of water, 1.9g of maltitol solution, 1.9g of honey, 1.3g of coconut oil, 0.6g of konjac powder, 0.6g of carrageenan, 0.01g of vanilla extract, 0.03g of potassium sorbate, 0.3g of whiteleg shrimp extract, and 0.7g of Gastrodia elata water extract.

[0010] Each 100g of the inner layer contains 21g of whey protein powder, 15g of oatmeal, 4g of soy protein powder, 2.5g of coconut oil, 1.7g of resistant dextrin, 1.7g of fructooligosaccharides, 0.03g of vitamin E, 1.7g of plain milk powder, cocoa powder, or matcha powder, 0.85g of erythritol, 0.85g of honey, 0.04g of vanilla extract, 2.5g of maltitol solution, 5g of plain milk powder, and 10g of water.

[0011] The *Litopenaeus vannamei* extract is a crustacean functional peptide prepared by enzymatic hydrolysis from *Litopenaeus vannamei* processing by-products, with a molecular weight below 1000 Daltons. The *Gastrodia elata* aqueous extract is an extract prepared by water extraction from the dried tubers of *Gastrodia elata* from Bomi region of Linzhi, Tibet. The aqueous extract contains at least one dominant polyphenolic substance selected from neohesperidin, forsythoside B, 4,4'-dihydroxystilbene, p-cresol, syringin, and raspberry ketone glucoside. The mass ratio of crustacean functional peptides to *Gastrodia elata* aqueous extract is 3:7.

[0012] A method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions includes the following steps:

[0013] Step 1 is to prepare the sandwich layer: Mix the extract of whiteleg shrimp, water extract of Gastrodia elata, whey protein powder, konjac powder, carrageenan, vanilla extract and potassium sorbate according to the formula, add water, maltitol solution, honey and coconut oil and mix well. Refrigerate for 30 minutes and set aside.

[0014] Step 2 is to prepare the inner layer: Mix whey protein powder, soy protein powder, plain milk powder, resistant dextrin, fructooligosaccharides, vitamin E, cocoa powder or matcha powder, and erythritol evenly. Add honey, vanilla extract, maltitol syrup, and water and knead. Add coconut oil until the dough is no longer sticky. Wrap the oat flakes evenly around the dough, shape it into a rectangle of 10cm×2cm×1cm, and bake for 16 minutes. The baking conditions are hot air baking at a temperature of 150-180℃. Set aside.

[0015] Step 3, shaping and post-processing: The filling layer obtained in step 1 is evenly spread on the outer surface of the inner layer obtained in step 2. Chocolate sauce is poured onto the outer surface of the filling layer and chopped nuts are sprinkled on top. After low-temperature sterilization and the addition of preservatives, the packaging is sealed to obtain the finished protein bar.

[0016] The preparation process of the extract of Litopenaeus vannamei, one of the core raw materials in the sandwich layer, includes the following steps:

[0017] S1: Raw material pretreatment: Remove impurities from the processing by-products of whiteleg shrimp, wash repeatedly with deionized water, drain the water, and freeze at -20℃.

[0018] S2: Ultrasonic and microwave extraction: Add deionized water at a material-to-liquid ratio of 3g:1mL, adjust the pH of the system to the optimal pH of the protease, and perform ultrasonic extraction at 400W for 30min and microwave extraction at 400W for 30min in sequence.

[0019] S3: Enzymatic hydrolysis and purification: Add 0.5% of protease and hydrolyze for 6 hours at the optimal pH and temperature of the protease. After inactivating the enzyme in a boiling water bath for 10 minutes, adjust the pH of the system to neutral. Centrifuge at 4°C and 10,000 r / min for 15 minutes. Filter to remove suspended solids and precipitates. Collect the supernatant and freeze-dry it under vacuum to obtain lyophilized crustacean functional peptide powder, namely the extract of Litopenaeus vannamei.

[0020] The protease mentioned in S3 is at least one of alkaline protease, papain, pepsin, and trypsin; wherein the optimal pH for enzymatic hydrolysis of alkaline protease is 8.0 and the optimal temperature is 50°C; the optimal pH for enzymatic hydrolysis of papain is 6.5 and the optimal temperature is 50°C; the optimal pH for enzymatic hydrolysis of trypsin is 7.5 and the optimal temperature is 37°C; and the optimal pH for enzymatic hydrolysis of pepsin is 2.0 and the optimal temperature is 37°C.

[0021] The preparation process of the water extract of Gastrodia elata, another core raw material of the sandwich layer, includes the following steps:

[0022] T1: Raw material pretreatment: Crush the dried tubers of Gastrodia elata from Bomi region of Linzhi, Tibet, and pass them through a 60-mesh sieve for later use;

[0023] T2: Water bath reflux extraction: Add deionized water at a material-to-liquid ratio of 1g:10mL, reflux in an 80℃ water bath for 2 hours, repeat the extraction twice, and combine the two extracts;

[0024] T3: Concentration and Drying: The combined extracts are concentrated under reduced pressure and then freeze-dried under vacuum to obtain freeze-dried powder of Tibetan Gastrodia elata water extract, namely the Tibetan Gastrodia elata water extract.

[0025] Four key control points are set during the preparation process:

[0026] During the raw material receiving stage, the heavy metal and microbial indicators of the raw materials are controlled to meet national standards;

[0027] During the sandwich preparation stage, the sandwich materials are monitored for microorganisms.

[0028] During the freeze-drying and shaping stage, the water activity Aw of the material should be controlled to be ≤0.55;

[0029] During the packaging stage, the residual oxygen content inside the packaging should be controlled to be ≤0.5%.

[0030] The low-temperature sterilization temperature is 45-60℃, and the sterilization time is 20-30 minutes. The instruments used in the preparation include an electronic balance, a vacuum freeze dryer, a constant temperature water bath, a mixer, a homogenizer, an oven, a freezer, a nitrogen-filling packaging machine, and a sealing machine.

[0031] The beneficial effects of this invention are as follows:

[0032] 1. The present invention discloses a protein bar with antioxidant and glucose and lipid metabolism regulation functions and its preparation method. This involves scientifically compounding crustacean functional peptides prepared by enzymatic hydrolysis of processing by-products of Litopenaeus vannamei with aqueous extracts of Gastrodia elata from Bomi, Linzhi, Tibet, at a mass ratio of 3:7. Combined with a synergistic design verified by the DPPH free radical scavenging method, this achieves a dual-functional enhancement of antioxidant and glucose and lipid metabolism regulation. After compounding, the DPPH free radical scavenging IC50 value is as low as 2.28 μg / mL, and the interaction index γ=0.57. The crustacean functional peptides have a significant effect on improving obesity induced by a high-sugar, high-fat diet, while the aqueous extract of Gastrodia elata can restore malondialdehyde content in cells damaged by acetaldehyde to normal levels. The synergistic effect of these two components compensates for the shortcomings of single functional ingredients, providing a more efficient nutritional regulation solution for sub-healthy individuals.

[0033] 2. The protein bar with antioxidant and glucose-lipid metabolism regulating functions and its preparation method described in this invention, through a three-layer structure formula design and low-temperature process control throughout the entire process, not only achieves the preservation of the activity of functional components, but also solves the problem of the fishy smell of shellfish raw materials; the sandwich layer is prepared at low temperature throughout the process, the core extract is freeze-dried in vacuum, and the finished product is sterilized at low temperature of 45-60℃, avoiding protein denaturation and degradation of active ingredients caused by high temperature. At the same time, the scientific combination of whey protein, oatmeal, chocolate, nut pieces and vanilla extract flavoring effectively neutralize the inherent fishy smell of shellfish raw materials. Attached Figure Description

[0034] The invention will now be further described with reference to the accompanying drawings.

[0035] Figure 1 This is a flowchart of the method of the present invention;

[0036] Figure 2 This is a schematic diagram illustrating the scavenging ability of different antioxidants of the present invention against DPPH free radicals;

[0037] Figure 3 This is a heatmap analysis diagram of the clustering relationship of compounds in Gastrodia elata according to the present invention;

[0038] Figure 4 This is a graph showing the average fluorescence intensity of reactive oxygen species (ROS) in this invention.

[0039] Figure 5 This is a bar graph showing the average fluorescence intensity of reactive oxygen species (ROS) in this invention.

[0040] Figure 6 These are photos of mice used to validate the animal model of this invention;

[0041] Figure 7 This is a schematic diagram illustrating the results of mouse weight change verification in the animal model of this invention. Detailed Implementation

[0042] The following will describe specific embodiments of the present invention and accompanying drawings. Figures 1-5 As shown, the technical solution of the present invention is clearly and completely described. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0043] A protein bar with antioxidant and glucose and lipid metabolism regulation functions has a three-layer composite structure consisting of an outer layer, a sandwich layer, and an inner layer. Each layer has an independent formula, and the amount of each ingredient added is determined based on 100g of its total weight. Each of the three layers has its own function: the inner layer is the nutritional base, the sandwich layer is the core functional layer, and the outer layer is the taste and shaping layer. The core functional ingredients are extracts of whiteleg shrimp and water extracts of Gastrodia elata, with a fixed mass ratio of 3:7, which is the key ratio to achieve synergistic antioxidant and glucose and lipid metabolism regulation.

[0044] Specifically, it includes:

[0045] Outer layer: Each 100g of outer layer contains 12g of chocolate and 4.8g of chopped nuts; the remainder consists of conventional base ingredients for chocolate products, such as cocoa butter and emulsifiers, which are common raw materials in the food industry and can be purchased directly from the market. The outer layer has no functional ingredients and its main function is to improve the product's formability and palatability, while slowing down the release rate of the inner and sandwich layer ingredients in the human body and prolonging the duration of their functional effects.

[0046] Filling layer: Each 100g of filling layer contains 6.5g whey protein powder, 2.5g water, 1.9g maltitol solution, 1.9g honey, 1.3g coconut oil, 0.6g konjac powder, 0.6g carrageenan, 0.01g vanilla extract, 0.03g potassium sorbate, 0.3g whiteleg shrimp extract, and 0.7g Tibetan gastrodia elata water extract. The filling layer is the core functional layer, which is concentrated with crustacean functional peptides and Tibetan gastrodia elata water extract. The remaining ingredients are shaping, flavoring, and preservative additives to ensure the stability of the filling layer's shape and taste.

[0047] Inner layer: Each 100g of inner layer contains 21g whey protein powder, 15g rolled oats, 4g soy protein powder, 2.5g coconut oil, 1.7g resistant dextrin, 1.7g fructooligosaccharides, 0.03g vitamin E, 1.7g plain milk powder, cocoa powder, or matcha powder, 0.85g erythritol, 0.85g honey, 0.04g vanilla extract, 2.5g maltitol liquid, 5g plain milk powder, and 10g water. The inner layer is the nutritional base layer and is the main source of high-quality protein and dietary fiber for the product. The option of adding "plain milk powder, cocoa powder, or matcha powder" is optional, meaning that one of them is added at 1.7g / 100g of the inner layer, corresponding to the plain, cocoa, and matcha flavors respectively, to meet diverse consumption needs.

[0048] Core functional ingredients and raw material characteristics:

[0049] All raw materials used in this invention are commercially available common raw materials or functional extracts prepared using conventional processes. There are no special or rare raw materials. The basic characteristics of each raw material and its role in this invention are as follows:

[0050] Litopenaeus vannamei extract: namely, crustacean functional peptides, are prepared by enzymatic hydrolysis using one or two of the processing by-products of Litopenaeus vannamei, namely shrimp heads and shells. With a molecular weight ≤1000 Daltons, it belongs to small molecule active peptides and has the characteristics of easy absorption and low allergenicity. It can regulate human glucose and lipid metabolism, scavenge free radicals to achieve basic antioxidant effects, and also play a role in immune regulation and anti-fatigue. Choosing shrimp heads and shells as raw materials can not only realize the high-value utilization of aquatic waste resources, but also reduce the raw material cost of the product.

[0051] Tibetan Gastrodia elata water extract: This product is made from dried tubers of Gastrodia elata from Bomi region of Linzhi, Tibet, using water extraction. This region is the authentic producing area of ​​Gastrodia elata, and its content of gastrodin and polyphenols is significantly higher than that of other producing areas. The extract contains at least one of the following polyphenolic substances with distinct advantages: neohesperidin, forsythoside B, 4,4'-dihydroxystilbene, p-cresol, syringin, and raspberry ketone glucoside. Its core function is to provide high-intensity antioxidant protection and improve cellular oxidative damage caused by exogenous toxic substances, while retaining the traditional effects of Gastrodia elata in calming the liver and relieving wind, and sedating the mind.

[0052] Whey protein powder, soy protein powder, and plain milk powder: Whey protein powder is a high-quality animal protein containing nutrients such as β-lactoglobulin and immunoglobulins, which are easily absorbed; soy protein powder is a high-quality plant protein that can reduce cholesterol intake; plain milk powder supplements protein and minerals such as calcium and zinc. The combination of the three achieves complementary effects between animal and plant proteins, avoids the nutritional deficiencies of single proteins, and provides the basic structure for protein bars.

[0053] Oatmeal, resistant dextrin, and fructooligosaccharides: Oatmeal is rich in beta-glucan, which increases satiety and slows down postprandial blood sugar; resistant dextrin is low in calories, increases satiety, and lowers blood sugar; fructooligosaccharides are prebiotics that regulate gut microbiota and cleanse the intestines and promote bowel movements. The combination of these three ingredients provides a diversified supplement of dietary fiber and helps regulate glucose and lipid metabolism.

[0054] Coconut oil: Its main component is medium-chain triglycerides, which can provide energy quickly without bile emulsification, are not easily converted into body fat, provide an appropriate amount of fat for the product, and at the same time improve the kneadability of raw materials and improve the taste of the product.

[0055] Maltitol, erythritol, and honey are all low-glycemic sweeteners that replace traditional refined sugar, preventing rapid spikes in blood sugar while providing a suitable level of sweetness. Honey also contains antioxidants such as flavonoids, which work in conjunction with the core functional ingredients to provide antioxidant support.

[0056] Konjac flour and carrageenan are both commonly used thickeners and molding agents in the food industry, which help maintain the stable shape of the sandwich layer and inner layer, making them less prone to breakage and deformation.

[0057] Vanilla extract: A general-purpose flavoring agent for food, it neutralizes minor off-flavors in raw materials and enhances the palatability of products.

[0058] Potassium sorbate: a general food preservative that inhibits microbial growth, extends product shelf life, and its addition amount complies with national food safety standards.

[0059] Vitamin E: A fat-soluble natural antioxidant that helps the core functional ingredients eliminate free radicals and enhances the overall antioxidant effect of the product.

[0060] Chocolate and chopped nuts: Common outer ingredients that enhance the product's texture and flavor, while the chopped nuts provide unsaturated fatty acids, enriching the product's nutrition.

[0061] A method for preparing protein bars with antioxidant and glucose-lipid metabolism regulating functions:

[0062] The preparation method of the protein bars of this invention has no special process requirements, and the instruments used are all conventional food processing equipment that can be purchased directly from the market. The entire preparation process is divided into four stages: self-made core functional ingredients, preparation of the sandwich layer, preparation of the inner layer, and molding and post-processing, as detailed below:

[0063] I. Preparations before preparation:

[0064] 1. Instrument Preparation: Electronic balance (accuracy 0.01g, for precise weighing of raw materials), vacuum freeze dryer (for drying and shaping of functional extracts), constant temperature water bath (for water extraction, enzymatic hydrolysis, and heat preservation), mixer (for mixing raw materials), homogenizer (for thorough kneading of raw materials), oven (for inner layer baking), freezer (for raw material preservation and sandwich layer refrigeration), nitrogen-filled packaging machine (for finished product packaging to prevent oxidation), sealing machine (for sealing packaging); all instruments should be cleaned with water and dried before use to avoid contamination of raw materials.

[0065] 2. Raw material preparation:

[0066] Commercially available raw materials: Whey protein powder, soy protein powder, oatmeal, coconut oil and other common raw materials are all selected from food-grade commercially available products and are used directly for preparation;

[0067] Homemade ingredients: Prepare extracts of whiteleg shrimp and water extract of Gastrodia elata in advance. Store in a freezer at -20°C after preparation. Thaw at room temperature before use.

[0068] II. Preparation of core functional ingredients:

[0069] 1. Preparation of extracts from Litopenaeus vannamei (crustacean functional peptides):

[0070] The ultrasonic-microwave-assisted enzymatic hydrolysis method is a routine method for extracting small molecule bioactive peptides in the food industry. The steps are detailed and the parameters are fixed, as follows:

[0071] S1: Raw material pretreatment: Take the processing by-products of whiteleg shrimp (one or two of shrimp heads and shells), manually remove impurities (such as mud, shrimp veins, etc.), wash repeatedly with deionized water 3-4 times, drain the surface water, and then freeze in a freezer at -20℃ for later use.

[0072] S2: Ultrasonic and microwave extraction: Weigh the pretreated shrimp heads / shells using an electronic balance, add deionized water to the raw materials at a material-to-liquid ratio of 3g:1mL, pour into the ultrasonic and microwave co-processing station, adjust the pH of the mixture to the optimal pH for the protease using 1mol / L sodium hydroxide solution or 1mol / L hydrochloric acid solution (adjust to the corresponding pH based on which protease will be used for subsequent enzymatic hydrolysis), turn on the ultrasonic function first, and extract with ultrasonic power at 400W for 30min. After ultrasonic extraction, do not change the container, directly turn on the microwave function, and extract with microwave power at 400W for 30min.

[0073] S3: Enzymatic hydrolysis reaction: Add 0.5% (m / m, i.e., the mass of the protease is 0.5% of the mass of the shrimp head / shell raw material) of protease to the mixture after ultrasonic and microwave extraction. The protease can be one or more of alkaline protease, papain, pepsin, and trypsin. After adding, stir evenly and hydrolyze at the optimal temperature and pH of the corresponding protease for 6 hours.

[0074] The optimal enzymatic hydrolysis conditions for each protease are: alkaline protease (pH 8.0, 50℃), papain (pH 6.5, 50℃), trypsin (pH 7.5, 37℃), and pepsin (pH 2.0, 37℃).

[0075] S4: Enzyme inactivation and purification: After 6 hours of enzymatic hydrolysis, pour the mixture into a beaker and heat it in a boiling water bath for 10 minutes to inactivate the enzyme at high temperature and prevent the protease from further decomposing the raw materials. After enzyme inactivation, adjust the pH of the mixture to neutral (pH 6.5-7.5) with 1 mol / L sodium hydroxide solution or 1 mol / L hydrochloric acid solution. Then pour the mixture into centrifuge tubes and place them in a refrigerated centrifuge. Centrifuge at 4°C and 10,000 r / min for 15 minutes. After centrifugation, take the supernatant and filter it with qualitative filter paper to remove suspended solids and precipitates to obtain the enzymatic hydrolysate.

[0076] S5: Drying and shaping: Pour the filtered enzymatic hydrolysate into the tray of a vacuum freeze dryer, turn on the equipment for vacuum freeze drying, and obtain a white or light yellow powdery solid after drying, which is the crustacean functional peptide freeze-dried powder, that is, the Litopenaeus vannamei extract of the present invention. After sealing the extract, freeze it at -20°C for later use.

[0077] 2. Preparation of water extract of Gastrodia elata from Tibet:

[0078] The water bath reflux extraction method is a conventional method for extracting water-soluble active ingredients from traditional Chinese medicinal materials, as detailed below:

[0079] T1: Raw material pretreatment: Take dried tubers of *Gastrodia elata* from Bomi region of Linzhi, Tibet, crush them with a pulverizer, and sieve them through a 60-mesh standard sieve to obtain Tibetan *Gastrodia elata* powder. After sieving, remove coarse residue and set aside. T2: Water bath reflux extraction: Weigh the Tibetan *Gastrodia elata* powder using an electronic balance, add deionized water at a material-to-liquid ratio of 1g:10mL, pour into a reflux extraction device, place the device in a constant temperature water bath, and reflux extract at 80℃ for 2 hours. After extraction, filter to obtain the first extract. Add deionized water again to the filter residue at a material-to-liquid ratio of 1g:10mL, and repeat the above 80℃ constant temperature reflux extraction operation to obtain the second extract. Combine the two extracts to obtain the total extract of Tibetan *Gastrodia elata*.

[0080] T3: Concentration and Drying: Pour the total extract of Gastrodia elata into a rotary evaporator and concentrate it under reduced pressure until the extract becomes a viscous paste. Pour the viscous paste into the tray of a vacuum freeze dryer and turn on the equipment for vacuum freeze drying. After drying, a light yellow powdery solid is obtained, which is the freeze-dried powder of Gastrodia elata water extract. After sealing, store it frozen at -20℃ for later use.

[0081] III. Preparation of the sandwich layer:

[0082] The sandwich layer is the core functional layer. The entire preparation process employs low-temperature treatment to avoid degradation of heat-sensitive components such as chitinous functional peptides and Tibetan gastrodia extract caused by high temperatures, thus preserving their bioactivity. Specific steps include:

[0083] Using an electronic balance, accurately weigh the following ingredients according to the formula: whiteleg shrimp extract, gastrodia elata water extract, whey protein powder, konjac powder, carrageenan, vanilla extract, and potassium sorbate. Pour all the dry ingredients into a mixer and mix at low speed for 5 minutes to ensure the dry ingredients are evenly mixed.

[0084] Add the prescribed amounts of water, maltitol syrup, honey, and coconut oil to the mixer, and continue mixing at low speed for 8-10 minutes until the ingredients are completely mixed and form a uniform, viscous paste.

[0085] Pour the paste into a food storage container, seal it, and refrigerate it at 4°C for 30 minutes to set the material and prevent it from flowing, thus obtaining the sandwich material. Refrigerate it for later use. The set sandwich material can be applied directly without further processing.

[0086] IV. Preparation of the inner layer:

[0087] The inner layer is the nutrient base layer. The core of its preparation lies in the thorough mixing and precise baking of the raw materials. The combination of grains and protein ingredients eliminates the fishy smell of the crustaceans. Specific steps:

[0088] Accurately weigh whey protein powder, soy protein powder, plain milk powder, resistant dextrin, fructooligosaccharides, vitamin E, and erythritol according to the formula using an electronic balance. Choose one of plain milk powder, cocoa powder, or matcha powder to add. Pour all dry ingredients into a homogenizer and homogenize at low speed for 3 minutes until well mixed.

[0089] Add the formula amount of honey, vanilla extract, maltitol solution and water to the homogenizer, turn on medium speed and homogenize for 5 minutes until the dry and wet ingredients are initially mixed to form a loose flocculent material;

[0090] Add the specified amount of coconut oil to the flocculent material and continue homogenizing at medium speed for 8-10 minutes, until the material is fully kneaded and forms a non-sticky dough (if the dough is too dry, add a small amount of water; if it is too wet, add a small amount of whey protein powder; these are standard adjustment methods in food processing).

[0091] Take out the kneaded dough and place it on the work surface. Manually wrap the oat flakes of the recipe evenly on the outer surface of the dough. Then, shape the dough into a standard rectangular shape of 10cm×2cm×1cm to obtain the inner raw dough. When shaping, ensure that the shape is regular and without cracks.

[0092] Arrange the inner layer dough evenly on a baking tray lined with parchment paper. Turn on the oven, set it to convection baking mode, and bake at 150-180℃ for 16 minutes. After baking, turn off the oven, remove the inner layer from the baking tray, and let it cool to 25-30℃ at room temperature to obtain the finished inner layer. Set aside after cooling.

[0093] V. Molding and Post-processing:

[0094] The final steps in making protein bars include coating the filling layer, preparing the outer layer, sterilization, and packaging. Key control points are also established to ensure product quality and safety. Specific steps are as follows:

[0095] Sandwich layer coating: Take the cooled inner layer product and use a scraper to evenly coat the refrigerated and shaped sandwich layer material onto the outer surface of the inner layer product. The coating thickness should be consistent (about 0.5-1cm) to avoid local areas being too thick or too thin, thus obtaining the sandwich inner layer.

[0096] Outer layer preparation: Melt the chocolate completely in a double boiler (heating temperature ≤60℃ to avoid chocolate spoilage). Use a brush to evenly spread the melted chocolate on the outer surface of the inner layer of the filling. Immediately after spreading, sprinkle chopped nuts on the chocolate surface. Sprinkle the chopped nuts evenly to form the outer layer. Then let it stand at room temperature for 10-15 minutes to allow the chocolate to solidify and obtain the semi-finished protein bar.

[0097] Low-temperature sterilization: Arrange the protein bar semi-finished product evenly in the sterilization tray, place it in the low-temperature sterilization equipment, set the sterilization temperature to 45-60℃, and the sterilization time to 20-30 minutes. Low-temperature sterilization is used instead of high-temperature sterilization. The core is to avoid the degradation of the functional components of the sandwich layer. After sterilization, take it out and let it cool at room temperature.

[0098] Adding preservatives and packaging: Spray a small amount of food-grade potassium sorbate solution (compliant with national standards) evenly onto the surface of the cooled protein bar semi-finished product. Then, place the protein bar into a food-grade sealed packaging bag and use a nitrogen-filling packaging machine to fill the packaging bag with nitrogen (to expel air and prevent product oxidation and deterioration). Immediately after filling with nitrogen, seal the packaging bag with a sealing machine to obtain the finished protein bar product.

[0099] Key Control Point Management: Four key control points must be strictly controlled throughout the preparation process:

[0100] Raw material receiving stage: Test all commercially purchased raw materials and self-made extracts for heavy metals and microbial indicators to ensure compliance with the National Food Safety Standards;

[0101] Sandwich preparation stage: Each batch of sandwich layer material undergoes microbial testing to prevent microbial contamination;

[0102] Freeze-drying and shaping stage: Test the water activity of the functional extract after freeze-drying to ensure Aw≤0.55 and prevent the extract from absorbing moisture and deteriorating;

[0103] Packaging stage: Test the residual oxygen content inside the packaging to ensure that the residual oxygen content is ≤0.5% and to guarantee the product's shelf life.

[0104] The protein bars of this invention achieve their function based on the characteristics of raw materials and the synergistic effect of components. The preparation process ensures the activity of functional components and the product's eating experience. From raw material selection and formula design to preparation and molding, each step works together to achieve a unity of nutrition, function, and taste. Even non-professionals can achieve the following working and technical effects after preparing the product according to the method of this invention.

[0105] The detailed work process and results are as follows:

[0106] The working process of this protein bar consists of two parts: the process coordination during preparation and the process of its effect on the human body after consumption. Both parts work together to ensure the product's functionality and nutritional value.

[0107] Process synergy in the preparation process:

[0108] When preparing the core functional ingredients, the ultrasonic and microwave-assisted enzymatic hydrolysis method can efficiently extract small molecule active peptides from shrimp heads or shells, and the water bath reflux extraction method can fully extract water-soluble antioxidant components from Tibetan Gastrodia elata. Both methods use vacuum freeze-drying to avoid the components from being deactivated due to high-temperature drying.

[0109] The sandwich layer is prepared and refrigerated throughout the entire process, the inner layer is baked at a temperature controlled at 150-180℃, and the finished product is sterilized at a low temperature of 45-60℃. The entire process avoids high temperatures and maximizes the preservation of the bioactivity of heat-sensitive components such as chitinous functional peptides, Tibetan gastrodia elata water extract, and vitamin E, laying the foundation for the realization of functions.

[0110] By scientifically combining ingredients such as whey protein powder, oatmeal, chocolate, and chopped nuts, and using vanilla extract for flavoring, the inherent fishy taste of crustaceans is effectively neutralized. At the same time, the three-layer structure design makes the product's taste rich and layered, avoiding the taste defects of a single ingredient.

[0111] Four key control points are managed throughout the entire process from raw materials to packaging, effectively avoiding the risks of heavy metal and microbial contamination and product oxidation, and ensuring stable product quality.

[0112] The process of human action after consumption:

[0113] After consuming the protein bar, during the chewing stage in the mouth, the outer layer of chocolate and chopped nuts quickly provides a pleasant flavor, while the inner layer of oatmeal and chopped nuts increases the chewiness and enhances the eating experience;

[0114] Once inside the stomach, the whey protein, soy protein, and original milk powder in the inner layer begin to break down under the action of gastric juice, quickly replenishing the body with high-quality amino acids to meet the needs of tissue repair and growth and development. Oatmeal, resistant dextrin, and fructooligosaccharides play a role in the intestines, increasing satiety, delaying the rise in blood sugar after meals, regulating intestinal flora, and helping to improve glucose and lipid metabolism.

[0115] The shell-derived functional peptides and the water extract of Gastrodia elata in the sandwich layer are small molecule components that are easily absorbed by the human intestines and enter the bloodstream. Among them, the shell-derived functional peptides directly act on the human glucose and lipid metabolism system, reduce the levels of triglycerides and cholesterol in the blood, improve metabolic disorders caused by high sugar and high fat, and at the same time clear a small number of free radicals in the body to achieve basic antioxidant effects. After entering the body, the water extract of Gastrodia elata enhances the body's antioxidant capacity by clearing a large number of free radicals in the body and increasing the expression of ALDH2 protein, thereby improving cellular oxidative damage caused by exogenous toxic substances such as alcohol and pollutants. At the same time, the polyphenols it contains form an antioxidant synergy with vitamin E and flavonoids in honey.

[0116] When chitosan functional peptides and Tibetan Gastrodia elata water extract are combined in a 3:7 mass ratio, they form a dual synergistic effect of antioxidation and regulation of glucose and lipid metabolism in the body. While regulating metabolism, the chitosan functional peptides provide basic antioxidants for Tibetan Gastrodia elata water extract. Tibetan Gastrodia elata water extract provides high-intensity antioxidants while protecting metabolic system cells from oxidative damage, significantly improving the overall functional effect. In addition, the outer chocolate layer slows down the absorption rate of the filling layer ingredients, allowing the functional ingredients to continue to work in the body and prolonging the duration of action.

[0117] Through the above-described formula design and preparation process, the protein bars produced by this invention not only achieve synergistic enhancement of core functions, but also solve several pain points of existing technologies. Furthermore, they possess advantages such as environmentally friendly raw materials, simple processing, and food safety. Specific technical effects are as follows:

[0118] Significant synergistic effects: When chitinous functional peptides and Tibetan Gastrodia elata water extract were combined in a 3:7 ratio, the IC50 value was as low as 2.28 μg / mL and the interaction index γ=0.57, as determined by the DPPH free radical scavenging method, showing a significant synergistic antioxidant effect, with antioxidant capacity far superior to that of the single component. Animal experiments confirmed that chitinous functional peptides can significantly improve obesity induced by a high-sugar, high-fat diet in mice, and Tibetan Gastrodia elata water extract can restore the malondialdehyde (MDA) content in cells damaged by acetaldehyde to normal levels. The combination of the two achieves dual functions of antioxidant and glucose and lipid metabolism regulation, with functional effects superior to existing single-function protein bars.

[0119] Achieving high-value utilization of aquatic waste resources: Functional peptides are prepared using the processing by-products (shrimp heads and shells) of whiteleg shrimp, which are discarded annually in quantities exceeding 10 million tons, as raw materials. This transforms aquatic waste into high-value-added functional raw materials, practices the concept of green circular economy, and reduces the cost of raw materials for products.

[0120] Promoting the food and modern development of Tibetan Gastrodia elata: Using Tibetan Gastrodia elata from Bomi County, Linzhi City, as raw material, and preserving its active ingredients through modern water extraction technology, this project integrates traditional Chinese medicine with functional foods, promotes the modernization of Tibetan medicine, and assists in industrial development in Tibetan areas, achieving synergistic upgrading of the fishery and Chinese medicine industries.

[0121] To maximize the preservation of the activity of functional ingredients: The entire process of low-temperature preparation of the sandwich layer, vacuum freeze-drying of the core extract, and low-temperature sterilization of the finished product is designed to avoid protein denaturation and degradation of functional ingredients caused by high temperature, thus ensuring the bioactivity of crustacean functional peptides and Tibetan Gastrodia elata water extract, and making the product's functional effects stable.

[0122] Excellent eating experience and high sensory score: Through scientific combination of raw materials and optimization of seasoning process, the inherent fishy smell of shellfish raw materials is effectively eliminated. The product has been rated by 100 professional sensory evaluators and has achieved a sensory score of 90 points. The three-layer structure design makes the taste rich and layered, and there are three flavors to choose from: original, cocoa and matcha, to meet the diversified consumer needs and achieve a high degree of unity between health benefits and eating experience.

[0123] Nutritionally comprehensive and suitable for a wide range of people: The product not only contains high-quality animal and plant protein and dietary fiber, but also contains minerals such as vitamin E, calcium, and zinc, as well as a variety of antioxidants. While achieving antioxidant and glucose and lipid metabolism regulation, it provides the human body with comprehensive and balanced nutrition, making it suitable for athletes, people in a sub-healthy state, people controlling their blood sugar, and general consumers.

[0124] The process is simple and easy to industrialize and prepare at home: All processes in this invention are conventional methods in the food industry, and all instruments used are commercially available equipment with no special customization requirements. The steps are detailed and the parameters are fixed, which can not only realize large-scale industrial production, but also allow non-professionals to prepare small batches at home by following the steps. Moreover, the raw materials are all food-grade commercial products that are easy to obtain.

[0125] Product quality and safety, long shelf life: Through full-process control of four key control points, risks such as heavy metal, microbial contamination and product oxidation are effectively avoided. The physicochemical indicators of the product, such as protein, fat and dietary fiber, meet national standards, and the microbial indicators meet the standards. Nitrogen-filled packaging effectively prevents product oxidation and deterioration, extending the product's shelf life.

[0126] Functional identification – animal model validation:

[0127] To verify the in vivo activity of chitinous functional peptides, a high-sugar, high-fat diet-induced obesity model was established using C57BL / 6 mice.

[0128] The experiment was divided into three groups: normal mice (normal diet), high sugar and high fat non-intervention group (high sugar and high fat diet), and high sugar and high fat intervention group (high sugar and high fat diet + chitinous functional peptide intervention).

[0129] After 21 weeks of continuous feeding and observation, the results were as follows: Figure 6 , Figure 7 As shown, a high-sugar, high-fat diet significantly induced weight gain in mice, while the weight of mice in the chitinous functional peptide intervention group was significantly lower than that in the non-intervention group at week 19, indicating that chitinous functional peptides have a significant ameliorative effect on obesity induced by a high-sugar, high-fat diet.

[0130] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of the present invention is defined by the appended claims and their equivalents.

Claims

1. A protein bar with antioxidant and glucose-lipid metabolism regulating functions, characterized in that, The protein bar has a three-layer structure, including an outer layer, a sandwich layer, and an inner layer; based on a total mass of 100g for each layer, the ingredients and amounts added to each layer are as follows: Each 100g of the outer layer contains 12g of chocolate and 4.8g of chopped nuts; Each 100g of the sandwich layer contains 6.5g of whey protein powder, 2.5g of water, 1.9g of maltitol solution, 1.9g of honey, 1.3g of coconut oil, 0.6g of konjac powder, 0.6g of carrageenan, 0.01g of vanilla extract, 0.03g of potassium sorbate, 0.3g of whiteleg shrimp extract, and 0.7g of Gastrodia elata water extract. Each 100g of the inner layer contains 21g of whey protein powder, 15g of oatmeal, 4g of soy protein powder, 2.5g of coconut oil, 1.7g of resistant dextrin, 1.7g of fructooligosaccharides, 0.03g of vitamin E, 1.7g of plain milk powder, cocoa powder, or matcha powder, 0.85g of erythritol, 0.85g of honey, 0.04g of vanilla extract, 2.5g of maltitol solution, 5g of plain milk powder, and 10g of water.

2. The protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 1, characterized in that, The whiteleg shrimp extract is a crustacean functional peptide prepared by enzymatic hydrolysis using whiteleg shrimp processing by-products as raw material. The Tibetan gastrodia elata water extract is an extract prepared by water extraction of the dried tuber of Gastrodia elata. The mass ratio of the crustacean functional peptide to the Tibetan gastrodia elata water extract is 3:

7.

3. A protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 2, characterized in that, The processing by-products of the whiteleg shrimp are shrimp heads and shells, and the molecular weight of the crustacean functional peptides is below 1000 Daltons.

4. The protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 2, characterized in that, The aqueous extract of Gastrodia elata contains at least one of the following polyphenolic substances with distinct advantages: neohesperidin, forsythoside B, 4,4'-dihydroxystilbene, p-cresol, syringin, and raspberry ketone glucoside.

5. A method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions, applicable to the preparation of a protein bar with antioxidant and glucose-lipid metabolism regulating functions as described in any one of claims 1-4, characterized in that, Includes the following steps: Step 1 is to prepare the sandwich layer: Mix the extract of whiteleg shrimp, water extract of Gastrodia elata, whey protein powder, konjac powder, carrageenan, vanilla extract and potassium sorbate according to the formula, add water, maltitol solution, honey and coconut oil and mix well. Refrigerate for 30 minutes and set aside. Step 2 is to prepare the inner layer: Mix whey protein powder, soy protein powder, plain milk powder, resistant dextrin, fructooligosaccharides, vitamin E, cocoa powder or matcha powder, and erythritol evenly. Add honey, vanilla extract, maltitol syrup, and water and knead. Add coconut oil until the dough is no longer sticky. Wrap the oatmeal evenly around the dough, shape it into a rectangle of 10cm×2cm×1cm, and bake for 16 minutes. Step 3, shaping and post-processing: The filling layer obtained in step 1 is evenly spread on the outer surface of the inner layer obtained in step 2. Chocolate sauce is poured onto the outer surface of the filling layer and chopped nuts are sprinkled on top. After low-temperature sterilization and the addition of preservatives, the packaging is sealed to obtain the finished protein bar.

6. The method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 5, characterized in that, The inner layer is prepared by hot air baking at a temperature of 150-180℃.

7. The method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 5, characterized in that, The preparation process of the Litopenaeus vannamei extract includes the following steps: S1: Raw material pretreatment: Remove impurities from the processing by-products of whiteleg shrimp, wash repeatedly with deionized water, drain the water, and freeze at -20℃. S2: Ultrasonic and microwave extraction: Add deionized water at a material-to-liquid ratio of 3g:1mL, adjust the pH of the system to the optimal pH of the protease, and perform ultrasonic extraction at 400W for 30min and microwave extraction at 400W for 30min in sequence. S3: Enzymatic hydrolysis and purification: Add 0.5% of protease and hydrolyze for 6 hours at the optimal pH and temperature of the protease. After inactivating the enzyme in a boiling water bath for 10 minutes, adjust the pH of the system to neutral. Centrifuge at 4°C and 10,000 r / min for 15 minutes. Filter to remove suspended solids and precipitates. Collect the supernatant and freeze-dry it under vacuum to obtain lyophilized crustacean functional peptide powder, namely the extract of Litopenaeus vannamei.

8. The method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 7, characterized in that, The protease mentioned in S3 is at least one of alkaline protease, papain, pepsin, and trypsin; wherein the optimal pH for enzymatic hydrolysis of alkaline protease is 8.0 and the optimal temperature is 50°C; the optimal pH for enzymatic hydrolysis of papain is 6.5 and the optimal temperature is 50°C; the optimal pH for enzymatic hydrolysis of trypsin is 7.5 and the optimal temperature is 37°C; and the optimal pH for enzymatic hydrolysis of pepsin is 2.0 and the optimal temperature is 37°C.

9. The method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 5, characterized in that, The preparation process of the Tibetan Gastrodia elata water extract Includes the following steps: T1: Raw material pretreatment: Crush the dried tubers of Gastrodia elata from Bomi region of Linzhi, Tibet, and pass them through a 60-mesh sieve for later use; T2: Water bath reflux extraction: Add deionized water at a material-to-liquid ratio of 1g:10mL, reflux in an 80℃ water bath for 2 hours, repeat the extraction twice, and combine the two extracts; T3: Concentration and Drying: The combined extracts are concentrated under reduced pressure and then freeze-dried under vacuum to obtain freeze-dried powder of Tibetan Gastrodia elata water extract, namely the Tibetan Gastrodia elata water extract.

10. A method for preparing a protein bar with antioxidant and glucose-lipid metabolism regulating functions according to claim 5, characterized in that, The low-temperature sterilization temperature is 45-60℃, and the sterilization time is 20-30 minutes.

Images