A continuous fluid extraction method for collagen peptides

Abstract

The present invention provides a continuous fluid extraction method for collagen peptides, which belongs to the technical field of natural protein peptide extraction. The present invention deashes and / or defattes animal raw materials, and the obtained material is heated and transferred to an enzymolysis tank after homogenization. At the same time, protease and pH value of the regulating system are added to the enzymolysis tank, and enzymolysis and filtration are performed in sequence. The filtered material is heated to the decolorization temperature during transportation, and fine filtration is performed after decolorization to obtain collagen peptides. The method provided by the present invention improves the utilization efficiency of equipment and saves equipment investment; the uniformity of auxiliary materials is improved by the auxiliary material flow addition mode; and the whole process time is shortened to 4 8h to obtain collagen peptide powder, which greatly reduces the risk of pollution, improves product quality, and achieves the purpose of saving production energy consumption.

Description

Technical Field

[0001] The invention belongs to the technical field of natural protein peptide extraction, and particularly relates to a continuous fluid extraction method for collagen peptide. Background Art

[0002] Bioactive peptides are a general term for diverse peptides composed of the 25 naturally occurring amino acids found in proteins in varying compositions and arrangements, ranging from dipeptides to complex linear and cyclic structures. These multifunctional compounds are derived from proteins. They possess diverse metabolic and physiological regulatory functions, are easily digested and absorbed, and have immune-boosting, hormone-regulating, antibacterial, antiviral, and blood pressure-lowering properties. They are highly safe for consumption, making them a hot research topic in the international food industry and a promising functional factor.

[0003] Bioactive peptides are primarily derived from proteins obtained from organisms, often recycled from processing byproducts. For example, active peptides are processed from fish skin, fish scales, cowhide, and soybean meal. Others are produced to enhance absorption, such as collagen peptides and sea cucumber peptides. Bioactive peptide extraction methods include water extraction, acid extraction, alkaline extraction, and enzyme extraction. Enzymatic hydrolysis is a relatively mature technology for extracting active peptides. The principle involves removing impurities from the target protein. Under certain parameters, such as the material-water ratio, temperature, enzyme dosage, pH, and time, large molecules are broken down into smaller pieces and dissolved in water. Insoluble matter is removed from the resulting product after the reaction is complete. The enzymatic hydrolyzate is then decolorized, deodorized, and refined, followed by concentration, sterilization, and drying. The aforementioned cleaning, crushing, enzymatic hydrolysis, filtration, decolorization, clarification, concentration, re-clarification, sterilization, and drying processes all require a reactor to contain the solution. Each batch of material proceeds to the next step after the reaction is complete. Each step involves ramping up and down times, waiting for the batch to be transferred to the tank, and time for the reaction to complete and be discharged. Often, the process from adding materials to drying and producing powder takes 24-48 hours, but the core processing time (enzymatic hydrolysis and decolorization) is only 2-3 hours. The rest of the time is spent waiting for material loading and unloading, or for temperature adjustment. This traditional solution has significant drawbacks: 1. It requires a large amount of equipment and high investment; 2. The longer the material is online, the greater the impact of heat treatment on color and odor; 4. The longer the online time, the greater the risk of bacterial contamination, which directly leads to product quality deterioration. Summary of the Invention

[0004] In view of this, the object of the present invention is to provide a continuous fluid extraction method for collagen peptides, which can significantly reduce material production time and ensure product quality.

[0005] The present invention provides a continuous fluid extraction method for collagen peptides, comprising the following steps: deashing and / or defatting an animal raw material to obtain a pretreated material;

[0006] homogenizing the pretreated material to obtain a homogenate;

[0007] The homogenate is heated to the enzymolysis temperature during the transfer process to the enzymolysis tank, and protease is added to the enzymolysis tank and the pH value of the system is adjusted. After the transfer is completed, the enzymolysis treatment is directly performed to inactivate the enzyme to obtain an enzymolysis product;

[0008] The enzymatic hydrolysis product is preliminarily filtered, and the preliminarily filtered material is heated to a decolorization temperature during transportation, and is decolorized and finely filtered to obtain collagen peptides.

[0009] Preferably, the animal raw material includes at least one of the following: animal skin, animal bones, animal tendons, animal shanks, fish scales and animal meat.

[0010] Preferably, the animal bones or fish scales include acid deashing and alkali defatting;

[0011] The animal skin, animal tendon, animal sinew or animal meat includes alkali defatting.

[0012] Preferably, the particle size of the material in the homogenate is comprised between 0.01 and 0.1 mm.

[0013] Preferably, the protease comprises at least one of the following: neutral protease, alkaline protease, papain and trypsin.

[0014] Preferably, the primary filtration or fine filtration is performed using one of the following instruments: a plate and frame filter, a candle filter, and a centrifuge.

[0015] Preferably, the initial filtration pressure of the plate and frame filter is 0.01 MPa, which is maintained for 15 minutes, and then the filtration pressure is increased to 0.1-0.15 MPa.

[0016] Preferably, the decolorization is completed by using activated carbon; the amount of activated carbon added is 5% to 30% of the solid content in the primary filter material;

[0017] During the decolorization process, activated carbon begins to be added during the transportation of the primary filter material.

[0018] Preferably, the fine filtration obtains a fine filtrate, and the process further comprises concentrating, sterilizing and drying the fine filtrate.

[0019] Preferably, the concentration method comprises using a nanofiltration membrane, a falling film double-effect or triple-effect concentration device or a scraper concentration device;

[0020] The sterilization is accomplished by pulsed intense light sterilization;

[0021] The drying includes spray drying; the air inlet temperature of the spray drying is 170-200°C, the air outlet temperature is 70-100°C, and the atomizer frequency is 30-50HZ.

[0022] The present invention provides a continuous fluid extraction method for collagen peptides, comprising the following steps: deashing and / or defatting an animal raw material to obtain a pretreated material; homogenizing the pretreated material to obtain a homogenate; heating the homogenate to an enzymatic hydrolysis temperature during transportation to an enzymatic hydrolysis tank, adding protease to the enzymatic hydrolysis tank and adjusting the pH value of the system; directly performing enzymatic hydrolysis after the transfer is completed, inactivating the enzyme, and obtaining an enzymatic hydrolysis product; performing a preliminary filtration on the enzymatic hydrolysis product, heating the obtained pre-filtered material to a decolorization temperature during transportation, and obtaining collagen peptides through decolorization and fine filtration. The present invention performs a deashing and / or defatting pretreatment on the animal raw material, which is beneficial for removing fat and ash from the animal raw material, reducing impurities, and laying the foundation for subsequent enzymatic hydrolysis. At the same time, the present invention performs homogenization after pretreatment of the animal raw material and before enzymatic hydrolysis, reducing the raw material volume, which is beneficial for shortening the enzymatic hydrolysis time and improving the uniformity of the enzymatic hydrolysis molecular weight distribution. In addition, the present invention also synchronizes the process of adjusting the enzymolysis temperature, pH value and adding protease during the material transfer process of the enzymolysis environment, adopts a flow-addition method, omits the time of adjusting the enzymolysis conditions after feeding, greatly shortens the overall duration of the preparation process, achieves the purpose of continuous extraction, thereby reducing the probability of browning of the product, and avoiding the product from being deteriorated and contaminated during long-term extraction. Experiments show that in the enzymolysis process, the traditional method is used to adjust the pretreated material to the appropriate conditions in the enzymolysis tank before adding enzyme solution for enzymolysis. Since the enzymolysis process involves heating and adjusting the pH value and feeding of a large volume of materials, it takes a long time, which greatly reduces the enzymolysis efficiency. The present invention, by synchronizing the adjustment of pH value, enzyme solution and heating measures during the material transportation process, can save material transportation time and the time of heating and adjusting pH after the end of transportation, greatly shorten the enzymolysis process time, and effectively improve the enzymolysis efficiency, thereby ensuring the quality of the enzymolysis product. It can be seen that the method provided by the present invention can greatly improve product quality by shortening the extraction cycle, ensuring that the prepared collagen peptide has good color, smell, and is not contaminated or deteriorated.

[0023] At the same time, the method provided by the present invention improves the utilization efficiency of the equipment and saves equipment investment; the material addition method is changed from the traditional per-tank addition method to the flow addition method, which improves the uniformity of the auxiliary materials; the entire process time is shortened to 4 to 8 hours to prepare collagen peptide powder; the process contamination risk is drastically reduced, and the product quality is better; and production energy consumption is saved.

[0024] Furthermore, the present invention specifically limits the particle size of the material in the homogenate to 0.01-0.1 mm, which is convenient for transportation and heating, has a small material particle size, high enzymatic hydrolysis uniformity, and saves energy. In addition, the present invention also specifically limits the sterilization to pulsed strong light sterilization technology, which is unobstructed at low temperature and ensures that the product color, smell and taste are optimal, no thermal reaction occurs, and unobstructed sterilization improves the material passing efficiency. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is an equipment composition diagram of the continuous fluid extraction method of collagen peptides implemented in the present invention. Note: 1: raw material pretreatment unit, 2: feeding unit, 3: meat grinder, 4: colloid mill, 5: conveying pipeline, 6: enzymatic hydrolysis tank, 7: plate and frame filter, 8: turnover tank, 9: concentration tower, 10: drying tower. DETAILED DESCRIPTION

[0026] The present invention provides a continuous fluid extraction method for collagen peptides, comprising the following steps:

[0027] Deashing and / or defatting the animal raw material to obtain a pretreated material;

[0028] homogenizing the pretreated material to obtain a homogenate;

[0029] The homogenate is heated to the enzymolysis temperature during the transfer process to the enzymolysis tank, and protease is added and the pH value of the system is adjusted. After the transfer is completed, the enzymolysis treatment is directly performed to inactivate the enzyme to obtain an enzymolysis product;

[0030] The enzymatic hydrolysis product is sequentially subjected to primary filtration, decolorization and fine filtration to obtain collagen peptides.

[0031] The present invention performs deashing and / or defatting treatment on animal raw materials to obtain pretreated materials.

[0032] In the present invention, the animal raw material preferably includes at least one of the following: animal skin, animal bones, animal tendons, animal shanks, fish scales, and animal meat. Preferred animal species include cattle, sheep, horses, donkeys, and mules. The fish include marine and / or freshwater fish. In the examples of the present invention, the continuous fluid extraction method for collagen peptides is illustrated using cattle bones, tilapia skin, and cod skin as examples.

[0033] In the present invention, the animal bones or fish scales preferably include acid deashing and alkaline degreasing. The animal skins, animal tendons, animal shanks or animal meat preferably include alkaline degreasing. The acid for the acid deashing is preferably an organic acid and / or an inorganic acid; for example, hydrochloric acid, sulfuric acid, phosphoric acid, citric acid. During the acid deashing, the volume concentration of the acid is preferably 1% to 5%, and can be 3.5%; the acid treatment time is preferably 12 to 48 hours, and can be 24 hours. The alkali in the alkaline degreasing includes sodium hydroxide or potassium hydroxide. During the alkaline degreasing, the mass concentration of the alkali is preferably 0.1% to 1%, and can be 0.5%; the alkali treatment time is preferably 12 to 72 hours, and can be 48 hours. The material after the treatment is washed to a pH value within the range of 7 to 9.

[0034] After obtaining the pretreated material, the present invention homogenizes the pretreated material to obtain a homogenate.

[0035] In the present invention, the homogenization is to gradually crush the pretreated material to form a homogenate with uniform particles. The crushing method includes a meat grinder and a colloid mill.

[0036] In the present invention, the particle size of the material in the homogenate is preferably 0.01 to 0.1 mm, preferably 0.02 to 0.08 mm, and may also be 0.04 to 0.07 mm, or 0.05 to 0.06 mm. The particles of the material in the homogenate are uniform.

[0037] After obtaining the homogenate, the present invention heats the homogenate to the enzymolysis temperature during the transportation process of transferring it to the enzymolysis tank, adds protease and adjusts the pH value of the system, and directly performs enzymolysis treatment after the transfer is completed to inactivate the enzyme and obtain an enzymolysis product.

[0038] In the present invention, the protease preferably includes at least one of the following: neutral protease, alkaline protease, papain and trypsin. It can be a combination of neutral protease and alkaline protease, a combination of neutral protease and papain, a combination of papain and trypsin, a combination of alkaline protease and papain, a combination of papain and trypsin, and a combination of alkaline protease and trypsin. It can also be a combination of neutral protease, alkaline protease and papain, a combination of neutral protease, alkaline protease and trypsin, a combination of alkaline protease, papain and trypsin, or a combination of neutral protease, alkaline protease, papain and trypsin. When the protease comprises more than two kinds, each protease is compounded according to the same enzyme activity. The added amount of the protease is 1500 to 2000 U / g. 蛋白 , or 1500U / g 蛋白 The pH of the system is preferably adjusted based on the optimal pH of each protease. For example, the optimal pH of neutral protease is 6.5-8; the optimal pH of alkaline protease is 9-11; the optimal pH of papain is 5-7; and the optimal pH of trypsin is 8-9. When a combination of proteases is used, each protease is preferably hydrolyzed in a stepwise manner. The hydrolysis temperature is preferably 50-60°C, and can be 55°C.

[0039] In the present invention, the enzyme inactivation method preferably includes high-temperature enzyme inactivation. The temperature of the high-temperature enzyme inactivation is preferably 90-98°C, and can be 95°C. The high-temperature enzyme inactivation time is preferably 5-15 minutes, and can be 10 minutes. The purpose of the enzyme inactivation is to inactivate the protease and avoid excessive enzymatic hydrolysis.

[0040] After obtaining the enzymatic hydrolysis product, the present invention sequentially subjects the enzymatic hydrolysis product to primary filtration, decolorization and fine filtration to obtain collagen peptides.

[0041] In the present invention, the primary filtration or fine filtration is preferably performed using one of the following instruments: a plate and frame filter, a candle filter, and a centrifuge. The initial filtration pressure of the plate and frame filter is preferably 0.01 MPa, maintained for 15 minutes, and then the filtration pressure is preferably increased to 0.1-0.15 MPa. The centrifugal speed of the centrifuge is preferably 3000-15000 rpm, and can be 4000-10000 rpm.

[0042] In the present invention, the decolorization is preferably performed using activated carbon. The amount of activated carbon added is preferably 5% to 30% of the solids in the primary filtration material, and can be 10%. The solution temperature during decolorization is preferably 50 to 90°C, and can be 70°C. The purpose of decolorization is to remove unnecessary impurities from the filtrate.

[0043] In the present invention, the fine filtration obtains the fine filtrate, and preferably further comprises concentrating, sterilizing and drying the fine filtrate.

[0044] In the present invention, the concentration method preferably includes the use of nanofiltration membrane, falling film double-effect or triple-effect concentration equipment or scraper concentration equipment. The concentration is preferably concentrated to 2 to 5 times the original volume. The sterilization is preferably completed by pulsed strong light sterilization method. The parameters of the pulsed strong light sterilization method are a pulse sterilizer with 30 lamps, a peak power of a single lamp greater than 1.5 MW, and a spectral range of 190 nm to 1200 nm. The drying preferably includes spray drying; the inlet air temperature of the spray drying is preferably 170 to 200 ° C, and can be 180 to 190 ° C; the outlet air temperature is preferably 70 to 100 ° C, and can be 80 ° C; the atomizer frequency is preferably 30 to 50 Hz, and can be 40 Hz.

[0045] In the present invention, after the drying, it is also preferably included in packaging and quality control. The packaging is preferably completed under aseptic conditions, and the packaging requirements are that the seal is firm, the seal line is straight, there is no empty seal, the bagging is accurately weighed, the label is clear, and the packaging materials are clean and tidy. The quality control includes metal detection, average molecular weight detection, sensory detection, color detection and other index detection. The qualified requirement of the metal detection is that metal foreign matter corresponding to the specifications of the metal detection test block shall not be detected. The average molecular weight detection includes detection in accordance with GB31645 standard. The color detection is preferably to configure a 10% aqueous solution of the prepared collagen peptide and measure the absorbance value at 420nm. The detection of other indicators includes detection according to GB31645 standard.

[0046] In the present invention, it is preferred to further provide a device for implementing the continuous fluid extraction method of collagen peptides, see Figure 1, comprising a raw material pretreatment unit, a feeding unit, a meat grinder, a colloid mill, a conveying pipeline, an enzymatic hydrolysis tank, a plate and frame filter, a turnover tank, a concentration tower, and a drying tower, which are sequentially connected by pipelines. The raw material pretreatment unit is used to pretreat animal raw materials. The raw material pretreatment unit is preferably an animal raw material soaking tank. The present invention has no special restrictions on the shape, specifications, and material of the soaking tank. The feeding unit is used to convey the pretreated animal raw materials to the meat grinder. The feeding unit is preferably a conveyor belt. The meat grinder and colloid mill are used to crush the animal raw materials to form a homogenate. The conveying pipeline is used to convey the raw slurry to the enzymatic hydrolysis tank, wherein the side wall of the conveying pipeline is also connected to the acid and alkali tank, the protease tank, and the heating device through pipelines, which are used to simultaneously add protease and pH adjustment reagents and heat the materials during transportation, thereby shortening the time for adjusting the pH value and temperature and adding protease after feeding, thereby shortening the fluid extraction cycle. The plate and frame filter is used for primary filtration and fine filtration. A heating jacket is also provided at the filter outlet of the plate and frame filter to heat the primary filter material to the decolorization temperature. The turnover tank is used for temporary storage and decolorization of the pre-filtered material to meet decolorization requirements. The concentration tower is used for concentration. The drying tower is used for vacuum drying. The device described in the present invention enables continuous extraction during collagen peptide preparation, significantly improving the degree of continuous extraction. Simultaneously, by synchronizing feeding, pH adjustment, protease addition, and enzymatic hydrolysis temperature, the entire extraction process cycle is greatly shortened, improving the utilization rate of each device, and increasing product yield within a certain period of time. Meanwhile, problems such as contamination, browning, and deterioration are avoided, significantly improving product quality.

[0047] The continuous fluid extraction method for collagen peptides provided by the present invention is described in detail below with reference to the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0048] Example 1

[0049] A continuous extraction method of collagen from tilapia skin

[0050] 1. Pretreatment: Using fresh tilapia skin as the raw material, add 0.5% sodium hydroxide by weight of the raw material, add water at a material-water ratio of 1:3, and process for 24 hours to degrease. After completion, wash the raw material to a pH of 8 and set aside.

[0051] 2. Crushing (homogenization): Mix the cleaned raw materials with water in a ratio of 1:5:1:1 (absolute dry protein (water removed from the raw materials)). Transfer the liquid to an emulsification tank and process it through an emulsification device (colloid mill, homogenizer) until the particle size in the slurry is 0.01 cm and there is no graininess to the touch. Before transportation, adjust the parameters of the heating jacket so that the outlet temperature of the heating jacket is 55°C and prepare to start transportation.

[0052] Here, the absolute dry protein content = protein content / (1-water content).

[0053] 3. Enzyme hydrolysis: After the start of transportation, neutral protease derived from Bacillus subtilis is added to the transportation pipeline (the filter port is equipped with a heating jacket) at a ratio of 1500U / g protein. The enzymatic hydrolysis reaction is carried out at 50°C for 60min (the reaction time starts from the material transportation time, the reaction time is greater than the transportation time, and the material stays in the enzymatic hydrolysis tank after transportation to reach the enzymatic hydrolysis time). After the end, the temperature is raised to 90°C and kept for 15min to complete the enzyme inactivation, thereby obtaining the enzyme-inactivated enzymatic hydrolysis product.

[0054] 4. Primary filtration: Clarify the enzymatic hydrolysate using a plate and frame filter, adjust the pressure of the plate and frame filter to 0.1-0.2 MPa; the pressure difference between the inlet and outlet shall not exceed 0.1 MPa to obtain the primary filtrate.

[0055] 5. Decolorization: Adjust the temperature of the heating jacket of the conveying pipe at the outlet of the filter so that the temperature of the primary filtered material after passing through the heating jacket rises to 60°C. Add activated carbon and acid for decolorization to the pipeline at the rear end of the heating jacket. The added mass of activated carbon is 5% of the solid content in the primary filtered material. Start counting 30 minutes from the start of carbon addition (filtration time is less than 30 minutes). When decolorization is completed, prepare for fine filtration.

[0056] 6. Fine filtration: Select a plate and frame filter for fine filtration. The plate and frame filter performs initial filtration at a pressure of 0.01 MPa. After maintaining for 15 minutes, gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate.

[0057] 7. Concentration: The refined filtrate is concentrated using a nanofiltration membrane, and the outlet discharge material concentration is 30% (solid mass percentage).

[0058] 8. Sterilization: Sterilize the concentrated liquid using a pulse sterilizer with 30 lamps, a single lamp with a peak power greater than 1.5 MW, and a spectral range of 190 nm to 1200 nm to obtain sterilized material.

[0059] 9. Drying: The sterilized material is spray-dried, and the air inlet temperature of the spray drying tower is controlled at 170°C, the air outlet temperature is controlled at 90°C, and the atomizer frequency is controlled at 23 Hz to 25 Hz to obtain tilapia skin-derived collagen peptide powder.

[0060] 10. Inner packaging: Packing is carried out under sterile conditions to obtain the finished product;

[0061] 12. Other product index testing

[0062] 1) Average molecular weight: the molecular weight distribution is determined according to the method in GB31645.

[0063] 2) Sensory evaluation: Prepare a sensory evaluation table (see Table 1) and evaluate by 5 people.

[0064] 3) Color: Determine by absorbance: Prepare a 10% by mass solution and measure the absorbance at 420 nm.

[0065] 4) Other physical and chemical indicators: in accordance with GB31645 standards.

[0066] Table 1 Sensory evaluation table

[0067]

[0068]

[0069]

[0070] result:

[0071] 1. The average molecular weight test results of the products are shown in Table 2. Small molecule peptides below 1000 Da account for more than 80%.

[0072] Table 2 Average molecular weight

[0073] Molecular weight range (Da) Peak area percentage (%, λ = 220nm) ＞5000 0 5000～3000 0.44 3000～2000 2.06 2000～1000 14.51 1000～500 35.5 500～180 44.13 ＜180 3.36

[0074] 2. Sensory evaluation

[0075] 5 people commented that the taste has no strange smell or bitterness, and the smell has a very slight characteristic flavor of collagen, without any strange smell.

[0076] 3. Color: OD 420nm is 0.06.

[0077] 4. Other physical and chemical indicators: in compliance with GB 31645. No metallic foreign matter was detected.

[0078] Example 2

[0079] A continuous extraction method of collagen from fresh cowhide

[0080] 1. Pretreatment: Using fresh cowhide as raw material, add 1% sodium hydroxide by weight, add water at a material-water ratio of 1:4, and degrease for 48 hours. Afterwards, wash the fresh cowhide with water to a pH of 8-9.

[0081] 2. Crushing (homogenization): The cleaned raw materials are fed into the emulsification tank at a material-to-water ratio of 1:10 based on the absolute dry protein. After being processed by the colloid mill, the slurry is obtained with a diameter of 0.02 cm and no granularity when touched. Before transportation, the heating jacket parameters are adjusted so that the slurry temperature at the heating jacket outlet is 50°C, and then transportation is ready;

[0082] 3. Enzymatic hydrolysis: After the start of transportation, the feed is heated to 1000U / g in the delivery pipe (after the outlet of the heating jacket). 蛋Alkaline protease derived from Bacillus subtilis was added to the mixture in a proportion of 4:1, and enzymatic hydrolysis reaction was carried out at 55°C for 30 minutes (the reaction time was counted from the material transportation time, the reaction time was greater than the transportation time, and the mixture stayed in a temporary storage tank after transportation to reach the enzymatic hydrolysis time). After the reaction, the temperature was raised to 95°C and kept for 15 minutes to inactivate the enzyme, thereby obtaining an enzyme-inactivated enzymatic hydrolysis product.

[0083] 4. Primary filtration: Use a centrifuge to clarify, centrifuge at a speed of 4000r / min, and separate the supernatant as the primary filtrate.

[0084] 5. Decolorization: Adjust the heating jacket of the conveying pipe at the outlet of the filter so that the temperature of the material reaches 70°C after passing through the heating jacket. Add activated carbon and acid for decolorization to the pipeline at the rear end of the heating jacket. The amount of activated carbon is 8% of the solid mass in the solution (solid mass = solution volume × solution concentration). Start timing 60 minutes from the start of carbon addition (filtration time is less than 60 minutes). When decolorization is completed, prepare for fine filtration.

[0085] 6. Fine filtration: Use a plate and frame filter for fine filtration. The initial filtration pressure of the plate and frame filter is 0.01MPa. After maintaining it for 15 minutes, gradually increase the filtration pressure to between 0.1MPa and 0.15MPa to obtain the fine filtrate.

[0086] 7. Concentration: The concentration method adopts falling film triple-effect concentration, and the outlet material concentration is 45% (solid mass percentage).

[0087] 8. Sterilization: The sterilization method is pulsed intense light sterilization technology. The specific parameters are a pulse sterilizer with 30 lamps. The peak power of a single lamp is greater than 1.5 mW, and the spectral range is 190nm to 1200nm.

[0088] 9. Spray drying: Spray drying is used as the drying method. The air inlet temperature of the spray drying tower is controlled at 170°C, the air outlet temperature is 70°C, and the atomizer frequency is 23HZ-25HZ to obtain collagen peptides derived from fresh cowhide.

[0089] 10. Inner packaging: Packaging should be carried out under sterile conditions; packaging requirements: firm seal, straight seal line, no empty seal, accurate bagging and weighing, clear labeling, and clean and tidy packaging materials.

[0090] 11. Metal detection: Metal foreign matter that corresponds to the specifications of the metal detection test block shall not be detected.

[0091] 12. The testing methods for other product indicators are the same as those described in Example 1.

[0092] result

[0093] 1. The average molecular weight of the products is shown in Table 3. Small molecule peptides below 1000 Da account for approximately 90%.

[0094] Table 3 Average molecular weight

[0095]

[0096]

[0097] 2. Sensory evaluation

[0098] 5 people commented that the taste has no strange smell or bitterness, and the smell has a very slight characteristic flavor of collagen, without any strange smell.

[0099] 3. Color: OD 420nm is 0.08.

[0100] 4. Other physical and chemical indicators: in compliance with GB 31645. No metallic foreign matter was detected.

[0101] Example 3

[0102] A continuous extraction method of collagen from fresh fish scales

[0103] 1. Pretreatment: Fresh fish scales are used as raw material. Deashing is performed by adding 1% hydrochloric acid (based on the raw material's weight). Water is added at a material-to-water ratio of 1:2.5. After 24 hours, the acid is washed off. Then, 1% sodium hydroxide (based on the raw material's weight) is added at a material-to-water ratio of 1:2.5. Water is added and the treatment is continued for 48 hours to defat. Afterwards, the fresh fish scales are rinsed with drinking water until the pH reaches 8-9.

[0104] 2. Crushing (homogenization): The cleaned raw materials are fed into the emulsification tank at a material-to-water ratio of 1:8 according to the absolute dry protein. After being processed through a series of emulsification equipment, the material size is 0.1cm and there is no granularity when touched. Before transportation, the parameters of the heating jacket are adjusted so that the slurry temperature at the heating jacket outlet is 60℃, and then transportation is ready;

[0105] 3. Enzymatic hydrolysis: After the start of transportation, the feed is heated to 2000U / g in the delivery pipe (after the outlet of the heating jacket). 蛋 Papain was added in a proportion of white to white, and the reaction was carried out for 90 minutes (the reaction time was counted from the material transportation time, the reaction time was greater than the transportation time, and the material stayed in a temporary storage tank after the transportation to achieve the enzymatic hydrolysis time). After the reaction, the temperature was raised to 95°C and kept for 10 minutes to inactivate the enzyme, thereby obtaining an enzyme-inactivated enzymatic hydrolysis product.

[0106] 4. Primary filtration: The enzymatic hydrolysis product after inactivation of enzymes is clarified by centrifuge at a speed of 4000 r / min and the supernatant is collected.

[0107] 5. Decolorization: Adjust the heating jacket of the conveying pipe at the outlet of the filter so that the temperature of the material reaches 65°C after passing through the heating jacket. Add activated carbon and acid for decolorization to the pipeline at the rear end of the heating jacket. The amount of activated carbon is 10% of the solid content in the solution. Start counting 60 minutes from the start of adding carbon (filtration time is less than 60 minutes). When decolorization is completed, prepare for fine filtration.

[0108] 6. Fine filtration: The decolorized liquid is finely filtered using a plate and frame filter. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining for 15 minutes, the filtration pressure is gradually increased to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate.

[0109] 7. Concentration: The refined filtrate was concentrated using a nanofiltration membrane with a pore size of 150 Da to make the feed concentration reach 25% (solid mass percentage) to obtain a concentrated solution.

[0110] 8. Sterilization: The concentrated liquid is sterilized by pulsed strong light sterilization technology. The specific parameters are a pulse sterilizer with 30 lamps, the peak power of a single lamp is greater than 1.5 mW, and the spectral range is 190 nm to 1200 nm to obtain a sterilized concentrated liquid.

[0111] 9. Drying: The sterilized concentrated liquid is spray-dried, and the air inlet temperature of the spray drying tower is controlled at 200°C, the air outlet temperature is controlled at 100°C, and the atomizer frequency is controlled at 25 Hz to obtain collagen powder derived from fresh fish scales.

[0112] 10. Inner packaging: Packaging should be carried out under sterile conditions; packaging requirements: firm seal, straight seal line, no empty seal, accurate bagging and weighing, clear labeling, and clean and tidy packaging materials.

[0113] 11. Metal detection: Metal foreign matter that corresponds to the specifications of the metal detection test block shall not be detected.

[0114] 12. The determination methods of other product indicators are the same as those described in Example 1.

[0115] result

[0116] 1. The average molecular weight of the products is shown in Table 4. Small molecule peptides below 1000 Da account for about 80%.

[0117] Table 4 Average molecular weight of the products

[0118] Molecular weight range (Da) Peak area percentage (%, λ = 220nm) ＞5000 0.1 5000--3000 0.16 3000--2000 2.11 2000--1000 16.7 1000--500 42.59 500--180 36.13 ＜180 2.21

[0119] 2. Sensory evaluation

[0120] 5 people commented that the taste has no strange smell or bitterness, and the smell has a very slight characteristic flavor of collagen, without any strange smell.

[0121] 3. Color: OD 420nm is 0.07.

[0122] 4. Other physical and chemical indicators: in compliance with GB 31645. No metallic foreign matter was detected.

[0123] Comparative Example 1

[0124] A continuous extraction method for collagen from fresh fish scales, without using a preparation process that requires simultaneous addition of materials and enzymes

[0125] 1. Pretreatment: Fresh fish scales are used as raw material. Deashing is performed by adding 1% hydrochloric acid (based on the raw material's weight). Water is added at a material-to-water ratio of 1:2.5. After 24 hours, the acid is washed off. Then, 1% sodium hydroxide (based on the raw material's weight) is added at a material-to-water ratio of 1:2.5. Water is added and the treatment is continued for 48 hours to defat. Afterwards, the fresh fish scales are rinsed with drinking water until the pH reaches 8-9.

[0126] 2. Enzyme hydrolysis: The cleaned fish scales are put into the enzymatic hydrolysis tank, heated to 60℃, and the enzyme is 2000U / g. 蛋白 Papain was added in a ratio of , and enzymatic hydrolysis was carried out for 90 min. After the end, the temperature was raised to 95°C and kept for 10 min to inactivate the enzyme, thereby obtaining an enzyme-inactivated enzymatic hydrolysis product.

[0127] 3. Primary filtration: The enzymatic hydrolysis product after enzyme inactivation is clarified by centrifuge at a speed of 4000 r / min, and the supernatant is collected and put into the decolorization tank.

[0128] 4. Decolorization: After the centrifugation of the entire tank of liquid is complete, heat the decolorized liquid to 65°C and add activated carbon to the supernatant, the amount of activated carbon being 10% of the substrate mass. Keep the temperature at 65°C for 60 minutes. After decolorization, prepare for fine filtration.

[0129] 5. Fine filtration: The decolorized liquid is finely filtered using a plate and frame filter. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining for 15 minutes, the filtration pressure is gradually increased to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate.

[0130] 6. Concentration: The refined filtrate is concentrated using a nanofiltration membrane with a pore size of 150D molecular weight to make the feed liquid concentration reach 25% to obtain a concentrated solution.

[0131] 7. Sterilization: The concentrated liquid is sterilized by pulsed strong light sterilization technology. The specific parameters are a pulse sterilizer with 30 lamps, a single lamp peak power greater than 1.5 mW, and a spectral range of 190 nm to 1200 nm to obtain a sterilized concentrated liquid.

[0132] 8. Drying: The sterilized concentrated liquid is spray-dried, and the air inlet temperature of the spray drying tower is controlled at 200°C, the air outlet temperature is controlled at 100°C, and the atomizer frequency is controlled at 25 Hz to obtain collagen powder derived from fresh fish scales.

[0133] 9. Inner packaging: Packaging should be carried out under sterile conditions; packaging requirements: the seal should be firm, the sealing line should be straight, there should be no empty seals, the bagging and weighing should be accurate, the labels should be clear, and the packaging materials should be clean and tidy.

[0134] 10. Metal detection: Metal foreign matter that corresponds to the specifications of the metal detection test block shall not be detected.

[0135] 11. The determination methods of other product indicators are the same as those described in Example 1.

[0136] result

[0137] 1. The average molecular weight of the products is shown in Table 5. Small molecule peptides below 1000 Da account for approximately 43%.

[0138] Table 5 Average molecular weight of the products

[0139] Molecular weight range (Da) Peak area percentage (%, λ = 220nm) ＞5000 7.28 5000--3000 15.9 3000--2000 13.01 2000--1000 20.43 1000--500 21.51 500--180 21.51 ＜180 0.37

[0140] 2. Sensory evaluation

[0141] 5 people commented: All of them felt that the taste was distinctly bitter and the smell had a distinct fishy smell.

[0142] 3. Color: OD 420nm It is 0.157.

[0143] 4. Other physical and chemical indicators: in compliance with GB 31645. No metallic foreign matter was detected.

[0144] Comparative Example 2

[0145] A continuous extraction method of collagen from tilapia skin

[0146] 1. Pretreatment: Using fresh tilapia skin as the raw material, add 0.5% sodium hydroxide by weight of the raw material, add water at a material-water ratio of 1:3, and process for 24 hours to degrease. After completion, wash the raw material to a pH of 8 and set aside.

[0147] 2. Enzyme hydrolysis: The cleaned fish skin is put into the enzymatic hydrolysis tank, heated to 50℃, and the enzyme is heated to 1500U / g. 蛋白 Neutral protease derived from Bacillus subtilis was added in a ratio of , and enzymatic hydrolysis was carried out for 60 min. After the end, the temperature was raised to 95°C and kept for 10 min to inactivate the enzyme, thereby obtaining an enzyme-inactivated enzymatic hydrolysis product.

[0148] 3. Primary filtration: Clarify the enzymatic hydrolysate using a plate and frame filter, adjust the pressure of the plate and frame filter to 0.1-0.2 MPa; the pressure difference between the inlet and outlet shall not exceed 0.1 MPa to obtain the primary filtrate.

[0149] 4. Decolorization: After the centrifugation of the entire tank of liquid is complete, heat the decolorized liquid to 60°C and add activated carbon to the supernatant, the amount of activated carbon being 5% of the substrate mass. Keep the temperature at 60°C for 30 minutes. After decolorization, prepare for fine filtration.

[0150] 5. Fine filtration: Select a plate and frame filter for fine filtration. The initial filtration of the plate and frame filter is at a pressure of 0.01 MPa. After maintaining for 15 minutes, gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain the fine filtrate.

[0151] 6. Concentration: The refined filtrate is concentrated using a nanofiltration membrane, with the outlet material concentration being 30% (solid mass percentage).

[0152] 7. Sterilization: Use pulsed strong light sterilization technology to sterilize the concentrate. The specific conditions are a pulse sterilizer with 30 lamps, a single lamp peak power greater than 1.5 mW, and a spectral range of 190 nm to 1200 nm to obtain sterilized materials.

[0153] 8. Drying: The sterilized material is spray-dried, and the air inlet temperature of the spray drying tower is controlled at 170°C, the air outlet temperature is controlled at 90°C, and the atomizer frequency is controlled at 23 Hz to 25 Hz to obtain tilapia skin-derived collagen peptide powder.

[0154] 9. Inner packaging: Packing is carried out under sterile conditions to obtain the finished product;

[0155] result:

[0156] 1. The average molecular weight test results of the products are shown in Table 6. Small molecule peptides below 1000 Da account for approximately 61%.

[0157] Table 6 Average molecular weight

[0158] Molecular weight range (Da) Peak area percentage (%, λ = 220nm) ＞5000 2.5 5000～3000 5.41 3000～2000 12.06 2000～1000 19.01 1000～500 30.51 500～180 28.21 ＜180 2.31

[0159] 2. Sensory evaluation

[0160] 5 people commented: All of them felt that the taste was distinctly bitter and the smell had a distinct fishy smell.

[0161] 3. Color: OD 420nm It is 0.132.

[0162] 4. Other physical and chemical indicators: in compliance with GB 31645. No metallic foreign matter was detected.

[0163] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A continuous fluid extraction method for collagen peptides, characterized in that: The following steps are involved: 1) Pretreatment: Fresh tilapia skin was used as raw material, sodium hydroxide (0.5% by weight of the raw material) was added, water was added according to a material-water ratio of 1:3, and the mixture was degreased for 24 hours; the raw material was washed until the pH value was 8; 2) Crushing: The cleaned raw materials are mixed with water in a mass ratio of 1:5 between absolute dry protein and water. The slurry is transferred to an emulsification tank and processed through an emulsification device to reduce the particle size of the slurry to 0.01 cm. Before transportation, the heating jacket parameters are adjusted to make the slurry temperature at the heating jacket outlet 55°C, and then transportation is ready to begin. 3) Enzymolysis: After the start of transportation, neutral protease derived from Bacillus subtilis was added to the transportation pipeline of the filter port equipped with a heating jacket at a ratio of 1500 U / g protein. The enzymatic hydrolysis reaction was carried out at 50°C for 60 minutes. The reaction time was measured from the material transportation time and the reaction time was greater than the transportation time. After the transportation was completed, the material was kept in a temporary storage tank for the enzymatic hydrolysis time. After the end of the enzymatic hydrolysis, the temperature was raised to 90°C and kept for 15 minutes to complete the enzyme inactivation, thereby obtaining the enzyme-inactivated enzymatic hydrolysis product. 4) Primary filtration: The enzymatic hydrolyzate is clarified using a plate and frame filter, and the pressure of the plate and frame filter is adjusted to 0.1-0.2 MPa to obtain a primary filtrate; 5) Decolorization: Adjust the temperature of the heating jacket of the conveying pipe at the filter outlet so that the temperature of the primary filtered material after passing through the heating jacket rises to 60°C. Add decolorization activated carbon and acid to the pipe at the rear end of the heating jacket. The amount of activated carbon added should be 5% of the solid content in the primary filtered material. Decolorization should be carried out for 30 minutes from the start of carbon addition. 6) Fine filtration: Use a plate and frame filter for fine filtration. Initially filter at a pressure of 0.01 MPa for 15 minutes, then gradually increase the filtration pressure to between 0.1 MPa and 0.15 MPa to obtain a fine filtrate. 7) Concentration: The refined filtrate is concentrated using a nanofiltration membrane, and the solid content by mass percentage of 30% in the material is discharged from the outlet to obtain tilapia skin-derived collagen peptides.

2. A continuous fluid extraction method for collagen peptides, characterized in that: The following steps are involved: S1) Pretreatment: Using fresh cowhide as raw material, adding 1% sodium hydroxide by weight of the raw material, adding water at a material-water ratio of 1:4, treating for 48 hours to perform degreasing, and washing the fresh cowhide with water to a pH value of 8-9; S2) Crushing: The cleaned raw materials are fed into an emulsification tank at a material-to-water ratio of 1:10 according to the absolute dry protein. The raw materials are processed by a colloid mill to obtain a slurry with a diameter of 0.02 cm. Before transportation, the parameters of the heating jacket are adjusted to make the slurry temperature at the outlet of the heating jacket 50°C, and then transportation is started; S3) Enzymatic hydrolysis: After the start of transportation, the transportation pipeline equipped with a heating jacket is heated at 1000 U / g. 蛋白 alkaline protease derived from Bacillus subtilis was added in a ratio of 1:1, and the enzymatic hydrolysis reaction was carried out at 55°C for 30 minutes. The reaction time was counted from the material transportation time, and the reaction time was greater than the transportation time. After the transportation, the material was kept in a temporary storage tank to reach the enzymatic hydrolysis time. After the end, the temperature was raised to 95°C and kept for 15 minutes to inactivate the enzyme, thereby obtaining an enzyme-inactivated enzymatic hydrolysis product; S4) Primary filtration: using a centrifuge to clarify the enzyme-inactivated enzymatic hydrolysate at a speed of 4000 r / min, and separating the supernatant as the primary filtrate; S5) Decolorization: Adjust the heating jacket of the delivery pipe at the filter outlet so that the temperature of the material reaches 70°C after passing through the heating jacket. Add decolorization activated carbon and acid to the pipe behind the heating jacket. The amount of activated carbon is 8% of the solids in the solution. Decolorization is carried out for 60 minutes from the start of carbon addition. Prepare for fine filtration. S6) Fine filtration: A plate and frame filter is used for fine filtration. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining it for 15 minutes, the filtration pressure is gradually increased to between 0.1 MPa and 0.15 MPa to obtain a fine filtrate. S7) Concentration: The concentration method adopts falling film triple-effect concentration, and the solid mass percentage of the outlet material is 45%, thereby obtaining collagen peptides derived from fresh cowhide.

3. A continuous fluid extraction method for collagen peptides, characterized in that: The following steps are involved: (1) Pretreatment: fresh fish scales are used as raw materials, hydrochloric acid accounting for 1% of the raw material weight is added for deashing, water is added according to a material-water ratio of 1:2.5, the acid solution is washed off after 24 hours, sodium hydroxide accounting for 1% of the raw material weight is added, water is added according to a material-water ratio of 1:2.5, the treatment is carried out for 48 hours, degreasing, and the fresh fish scales are washed with domestic water until the pH value is 8-9; (2) Crushing: The cleaned raw materials are fed into the emulsification tank at a material-water ratio of 1:8 according to the absolute dry protein, and processed through a series of emulsification equipment. The material size is 0.1 cm. Before transportation, the parameters of the heating jacket are adjusted so that the slurry temperature at the outlet of the heating jacket is 60 ° C, and then transportation is ready; (3) Enzymolysis: After the start of transportation, the delivery pipe with a heating jacket is set at the discharge port according to 2000U / g 蛋白 Add papain in a ratio of 100mg / L and react for 90 minutes. The reaction time starts from the material transportation time and the reaction time is longer than the transportation time. After the transportation is completed, the material is kept in a temporary storage tank to achieve the enzymatic hydrolysis time. After the end, the temperature is raised to 95℃ and kept for 10 minutes to inactivate the enzyme to obtain the enzyme-inactivated enzymatic hydrolysis product. (4) Primary filtration: The enzymatic hydrolysate after inactivation of enzymes was clarified using a centrifuge at a speed of 4000 r / min and the supernatant was collected; (5) Decolorization: Adjust the heating jacket of the conveying pipe at the outlet of the filter so that the temperature of the material reaches 65°C after passing through the heating jacket. Add activated carbon and acid for decolorization to the pipe at the rear end of the heating jacket. The amount of activated carbon should be 10% of the solid content in the solution. Start counting from the time of adding carbon. If the filtration time is less than 60 minutes, the decolorization is completed and the fine filtration is ready. (6) Fine filtration: The decolorized liquid is finely filtered using a plate and frame filter. The initial filtration pressure of the plate and frame filter is 0.01 MPa. After maintaining for 15 minutes, the filtration pressure is gradually increased to between 0.1 MPa and 0.15 MPa to obtain a fine filtrate. (7) Concentration: The refined filtrate was concentrated using a nanofiltration membrane with a pore size of 150 Da to reduce the solid content to 25%, thereby obtaining collagen peptides derived from fresh fish scales.

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