Microcapsule seasoning powder containing active plant components for cats and preparation method of microcapsule seasoning powder

Abstract

The invention provides microcapsule seasoning powder containing active plant components for cats and a preparation method of the microcapsule seasoning powder, the microcapsule seasoning powder comprises a double-layer embedding system, and the microcapsule seasoning powder comprises the following raw material components in percentage by weight: 10-14 wt% of a composite core material, 32-36 wt% of a multifunctional food calling base material, 35-39 wt% of the double-layer embedding system and 14-16 wt% of functional auxiliary materials; the composite core material is prepared by compounding a mint extract, a green tea extract, a chicory extract, a mulberry leaf extract and a hawthorn extract according to the mass ratio of (1.8-4.5): (1.2-3): (0.8-1.5): (0.5-1.2): (1-2). The cat food has a dual-stage targeting effect of low release in the stomach and high release in the intestinal tract, has high palatability, has the effects of promoting the health of the intestinal tract, improving metabolism and resisting aging synergistically, is long-acting and stable, and can be suitable for various types of functional foods for cats.

Description

Technical Field

[0001] This invention belongs to the field of pet food, and in particular relates to a microcapsule flavoring powder for cats containing active plant ingredients and its preparation method. Background Technology

[0002] Currently, cat flavoring powders are transitioning from "single palatability enhancement" to "palatability enhancement and health synergy." Core technologies focus on four aspects, but all have shortcomings: While palatability enhancers are shifting from traditional animal sources (chicken liver powder, fish paste) or chemically synthesized flavorings to natural plant extracts (peppermint, green tea), they are mostly single additions lacking functional synergy; active ingredient encapsulation primarily uses single-layer wall materials (gum arabic, gelatin), and a few double-layer encapsulations cannot adapt to the cat's digestive tract due to the lack of pH sensitivity in the inner layer; functional design focuses on single needs (such as probiotics for gut health, tea polyphenols for antioxidant effects), neglecting component synergy (such as prebiotic-probiotic, polyphenol-flavonoid) and the balance between "function and palatability"; freshwater fish enzymatic hydrolysis often uses single enzymes, resulting in low efficiency and a lack of end-to-end stability control, leading to low retention rates of active ingredients.

[0003] Existing technologies still have four major drawbacks: insufficient targeted release capability, the wall material is easily degraded in the stomach, causing the active ingredients (probiotics, polyphenols) to be destroyed by gastric acid, and the intestinal release rate is less than 80%; weak synergistic effect of ingredients, single function (such as only attracting appetite or only regulating the intestines), and some high-concentration functional ingredients can reduce palatability, causing a decrease in feed intake of more than 20%; poor process and adaptability, single enzymatic hydrolysis results in a protein hydrolysis degree of less than 40%, and the activity retention rate is less than 75% after 6 months of storage at room temperature, and there is no differentiated design for senior cats, obese cats, etc.; some products contain chemically synthesized palatability attractants or low-purity raw materials, which pose a safety risk of irritating the intestines or causing soft stools.

[0004] Therefore, there is an urgent need for cat microcapsule flavoring powder containing active plant ingredients and its preparation method to solve the core pain points of existing products such as "poor targeting, single function, insufficient stability and low compatibility". Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a microcapsule flavoring powder for cats containing active plant ingredients and its preparation method. Through a double-layer encapsulation system, a five-element plant composite core material, and a two-step composite enzymatic hydrolysis palatability-inducing base material, it achieves a dual-stage targeted effect of low release in the stomach and high release in the intestine. It also has high palatability, promotes intestinal health, improves metabolism, and has synergistic anti-aging effects. Furthermore, it is long-lasting and stable, and can be adapted to various types of functional cat foods.

[0006] This invention provides a microcapsule flavoring powder for cats containing active plant ingredients, comprising a double-layer encapsulation system. By weight percentage, the raw material components are: 10-14 wt% composite core material, 32-36 wt% multifunctional palatability-enhancing base material, 35-39 wt% double-layer encapsulation system, and 14-16 wt% functional excipients. The composite core material is composed of peppermint extract, green tea extract, chicory extract, mulberry leaf extract and hawthorn extract in a mass ratio of (1.8-4.5):(1.2-3):(0.8-1.5):(0.5-1.2):(1-2); The multifunctional attractant base is composed of freshwater fish enzymatic hydrolysate, brewer's yeast extract, compound amino acids and natural attractants in a mass ratio of (4-7):(1.5-3):(1-2):(0.05-0.15); The bilayer encapsulation system comprises an inner nanocarrier and an outer microcapsule wall material. The inner nanocarrier is a mixture of sodium alginate-chitosan complex and liposome-encapsulated L-carnosine, with a mass ratio of 3-5:1 and a particle size of 50-100 nm. The outer microcapsule wall material is a mixture of gum arabic, gelatin, glycerol, and β-cyclodextrin, with a mass ratio of 2.5-4.5:1.5-3.5:0.8-1.8:0.5-1.2 and a particle size of 10-25 μm. The functional excipients include maltodextrin, a cryoprotectant, thermoresistant probiotics, sodium dihydrogen phosphate, and tert-butylhydroquinone, which are mixed in a mass ratio of (8.5-18):(3-8):(0.5-2):(0.8-2.2):(0.015-0.05). The cryoprotectant is composed of trehalose and mannitol in a mass ratio of 1:1-3:1.

[0007] Furthermore, each extract in the composite core material meets the following quality indicators: The menthol content in the peppermint extract is ≥18 wt%; The tea polyphenol content in green tea extract is ≥25wt%; The inulin content in chicory extract is ≥60wt%; The total flavonoid content in mulberry leaf extract is ≥30wt%; The triterpenic acid content in hawthorn extract is ≥15wt%.

[0008] Furthermore, the multifunctional palatability-enhancing substrate includes: The degree of protein hydrolysis of freshwater fish enzymatic hydrolysate is ≥48%, and the solid content is 28-32 wt%. The total content of disodium 5'-inosinate and disodium 5'-guanylate in brewer's yeast extract is ≥4 wt%; The compound amino acid is composed of taurine, food-grade L-glycine, and glutamic acid in a mass ratio of (2-3):(3-4):(1-1.5), with a total purity ≥95wt%, of which taurine purity ≥98wt%. The natural attractant is selected from salmon hydrolyzed peptides or taurine with a purity of ≥99wt%, wherein the molecular weight of the salmon hydrolyzed peptides is 500-1000 Da.

[0009] Furthermore, the double-layer embedding system includes: The mass ratio of sodium alginate to chitosan in the inner nanocarrier is 3:1-5:1, and the amount of cross-linking agent CaCl2 added is 0.5-1.5wt%; the mass ratio of L-carnosine encapsulated by liposomes is L-carnosine: dipalmitoylphosphatidylcholine = 1:1, the liposome particle size is 50-100nm, and the encapsulation efficiency is ≥95%; In the outer microcapsule wall material, the viscosity of gum arabic in a 10% aqueous solution at 25℃ is 220-320 mPa·s, the gel strength of gelatin is 230-260 Bloom and the ash content is ≤1.2wt%, the purity of glycerol is ≥99.5%, and the purity of β-cyclodextrin is ≥99% and the moisture content is ≤10wt%. The total encapsulation efficiency of the double-layer encapsulation system for the composite core material and L-carnosine is ≥92%.

[0010] Furthermore, the functional excipients include: Maltodextrin has a DE value of 12-18 and a moisture content of ≤5 wt%. The thermoresistant probiotic is Lactobacillus plantarum, with a survival rate of ≥90% and a viable count of ≥1×10⁻⁶ after being treated at 60℃ for 72 hours. 9 CFU / g; Sodium dihydrogen phosphate purity ≥99%, the pH of the system can be adjusted to 6.0-6.5; The amount of tert-butylhydroquinone added is 0.018-0.05 wt% of the total mass of the flavoring powder.

[0011] Furthermore, the dual-layer encapsulation system achieves dual-stage targeted release: the total release rate of the composite core material and L-carnosine within 2 hours under simulated cat gastric environment is ≤10%, and the total release rate of the composite core material and L-carnosine within 4 hours under simulated cat intestinal environment is ≥92%, and the difference between the intestinal release rate and the gastric release rate is ≥82%.

[0012] Furthermore, the microcapsule particle size of the seasoning powder is 10-25 μm; the moisture content of the seasoning powder is ≤5%, and the solubility at 25℃ is ≥92%; after storage at 25℃ and 65% relative humidity for 6 months, the retention rate of menthol is ≥88%, the retention rate of tea polyphenols is ≥85%, the retention rate of L-carnosine is ≥82%, and the survival rate of probiotics is ≥85%; after an accelerated test at 60℃ for 30 days, the retention rate of each active ingredient is ≥85%.

[0013] This invention provides a method for preparing a cat-specific microcapsule flavoring powder containing active plant ingredients, comprising the following steps: S1. Preparation of composite core material: Peppermint extract, green tea extract, chicory extract, mulberry leaf extract, and hawthorn extract are compounded in a mass ratio of (1.8-4.5):(1.2-3):(0.8-1.5):(0.5-1.2):(1-2) and mixed evenly to obtain composite core material; wherein the menthol content of peppermint extract is ≥18wt%, the tea polyphenol content of green tea extract is ≥25wt%, the inulin content of chicory extract is ≥60wt%, the total flavonoid content of mulberry leaf extract is ≥30wt%, and the triterpenic acid content of hawthorn extract is ≥15wt%. S2. Construction of the inner nanocarrier: ① Prepare L-carnosine liposomes by mixing L-carnosine and dipalmitoylphosphatidylcholine at a mass ratio of 1:1 and dispersing them in PBS buffer at pH 7.4; ② Mix 2% sodium alginate solution and 1% chitosan solution at a mass ratio of 3-5:1, add 0.5-1.5wt% CaCl2 solution, and solidify by electrostatic adsorption reaction at 30℃ for 30±2min to obtain composite microspheres; ③ Add the composite core material and L-carnosine liposomes to the composite microspheres at a mass ratio of 1:(0.3-0.5), and stir at 35-40℃ and 120-150r / min for 30-40min to obtain the nanocarrier-active ingredient complex; S3. Preparation of outer microcapsule wall material: Weigh gum arabic, gelatin, and glycerin according to the proportion, add deionized water, stir at 65-72℃ until dissolved, cool to 38-42℃, add β-cyclodextrin, stir for 25-30 min to obtain wall material composite solution, the solid content of the wall material composite solution is 18-22 wt%; S4. Double-layer embedding and spray drying: The nanocarrier-active ingredient complex is added dropwise to the wall material composite solution at a volume ratio of 1:(2.5-3.5), stirred at 38-42℃ and 80-100 r / min for 1.2-1.8 h, the pH is adjusted to 4.2-4.6, and stirring is continued for 40-50 min to obtain a suspension; the suspension is spray dried with the following parameters: inlet air temperature 190-210℃, outlet air temperature 85-92℃, feed rate 18-22 mL / min, atomization pressure 0.25-0.35 MPa, and the dried powder is collected; S5. Mixing of excipients: Add functional excipients to the dry powder and stir at 120-150 r / min for 18-25 min to obtain cat microcapsule flavoring powder containing active plant ingredients.

[0014] Furthermore, in step S4, the inlet air temperature of the spray drying is controlled at 195-205℃ and the outlet air temperature is controlled at 87-90℃; in step S5, the stirring speed is adjusted to 130-140 r / min and the stirring time is extended to 22-25 min.

[0015] This invention provides an application of a microcapsule flavoring powder for cats containing active plant ingredients. The flavoring powder is added at an amount of 1.5-3% of the total mass of the cat functional food to prepare the cat functional food. The cat functional food is selected from adult cat extruded dry food, senior cat low-temperature baked dry food, obese cat low-fat canned food, or postoperative cat nutritional freeze-dried meat strips. The shelf life of the cat functional food is 12 months under the conditions of 25°C and 65% relative humidity.

[0016] The advantages and positive effects of this invention are: 1. The double-layer encapsulation system enables precise targeted release, with a total release rate of ≤10% of active ingredients within 2 hours in the simulated feline gastric environment and ≥92% within 4 hours in the intestinal environment, effectively avoiding gastric acid damage and significantly improving the utilization rate of active ingredients; 2. Synergistic effect of multiple ingredients: The five-element plant composite core material, combined with multifunctional palatability-enhancing base material and heat-resistant probiotics, not only ensures a feed preference rate of ≥92%, but also increases the number of bifidobacteria in cat feces by 25-35%, taking into account both palatability and intestinal health. 3. The product has excellent stability. The active ingredient retention rate is ≥82% after 6 months of storage at 25℃ / 65%RH, and ≥85% after 30 days of accelerated testing at 60℃. It is suitable for various types of cat functional foods and has a shelf life of up to 12 months. Detailed Implementation

[0018] Example 1: Preparation of Standard Formulation I. Raw Material Preparation and Processing 1. Preparation process of peppermint extract 1.1 Raw material pretreatment: Fresh spearmint stems and leaves were selected and processed within 24 hours after harvesting, with an initial moisture content of ≤8wt%. After manually removing withered leaves, weeds, and other impurities, the raw materials were washed three times with deionized water, each time for 10 minutes. After washing, the materials were placed in a forced-air drying oven and dried at 60℃ until the moisture content dropped to 5wt%. The dried materials were then pulverized and passed through a 40-mesh sieve to remove coarse fibers. The sieve-passing material was collected to obtain 820g of spearmint powder.

[0019] 1.2 Supercritical CO2 Extraction: Peppermint powder was loaded into a 500mL HA221-50-06 supercritical extraction vessel. The extraction pressure was set to 28MPa, the extraction temperature to 45℃, and the CO2 flow rate to 20L / h. The extraction process was carried out in two steps: first, static extraction for 1h (CO2 flow was stopped to ensure sufficient contact between the raw material and the solvent), and then dynamic extraction for 2h (CO2 was continuously introduced), for a total extraction time of 3h.

[0020] 1.3 Separation and Concentration: The extract was collected in a two-stage separation vessel. The first-stage separation conditions were 8 MPa and 50 °C, and the second-stage separation conditions were 5 MPa and 40 °C. The collected extract was transferred to a rotary evaporator and concentrated to a solid content of 30 wt% at 60 °C and -0.09 MPa, finally yielding 125 g of dark green oily peppermint extract.

[0021] 1.4. Quality testing: The menthol content was determined using an Agilent 1260 high-performance liquid chromatography system with a mobile phase of methanol-0.1% phosphoric acid water = 60:40, a flow rate of 1.0 mL / min, a detection wavelength of 254 nm, and a column temperature of 30 ℃. The measured menthol content was 21.3 ± 0.5 wt% (the required value is ≥18 wt%). Gas chromatography-mass spectrometry (GC-MS) confirmed the absence of solvent residue.

[0022] 2. Preparation of green tea extract 2.1 Water extraction process: Take 0.5 kg of Grade 1 green tea dried product with a moisture content ≤ 6 wt%; after crushing the green tea dried product, pass it through a 20-mesh sieve, add 7.0 L of deionized water at a material-to-liquid ratio of 1:14, and extract twice in a constant temperature water bath at 88℃ with stirring, each extraction lasting 1.5 h; after extraction, filter with a 100-mesh filter cloth, combine the filtrates obtained from the two filtrations, and finally obtain 12.5 L of filtrate.

[0023] 2.2 Purification with AB-8 macroporous resin (1) Resin pretreatment: Take 500mL of AB-8 resin, add 95% ethanol and soak for 24h, ensuring that the ethanol level is 5cm above the resin; after soaking, wash with deionized water until there is no alcohol smell, and confirm that there is no ethanol residue by testing with an alcohol meter; then soak in 5% hydrochloric acid and 5% sodium hydroxide for 4h each, and wash with deionized water after each soaking until the solution is neutral to complete the resin pretreatment.

[0024] (2) Sample loading and elution: The filtrate obtained from water extraction was loaded onto the pretreated AB-8 resin column at a flow rate of 2.5 BV / h (BV is the resin volume); after the sample loading was completed, it was eluted with 3 times the resin volume of deionized water until the eluent was colorless; then it was eluted with 40% ethanol at a flow rate of 2.5 BV / h, and the eluent was collected, finally yielding 3.2 L of eluent.

[0025] 2.3 Concentration and Drying: The collected eluent was transferred to a rotary evaporator and concentrated to a solid content of 28wt% at 70℃ and -0.08MPa. The concentrate was then transferred to a vacuum freeze dryer and dried at -50℃ and 0.01MPa. After drying, 86g of pale yellow green tea extract powder was obtained.

[0026] 2.4 Quality Testing: The content of tea polyphenols was determined using the Folin-Ciocalteu method with gallic acid as the standard. The tea polyphenol content was found to be 28.5 ± 0.7 wt%, meeting the requirement of ≥25 wt%. The synergistic effect was verified by accelerated oxidation test: when the green tea extract and mulberry leaf extract were mixed at a mass ratio of 2.1:0.9, the oxidation half-life of total mulberry leaf flavonoids was extended from 12 h to 32 h, an extension factor of 2.67 times, meeting the synergistic requirement of ≥2.5 times.

[0027] 3. Preparation process of chicory extract 3.1 Raw material processing: Take 0.2 kg of dried chicory root, which should be free of mold and have an initial inulin content of ≥45 wt%; cut the chicory root into uniform thin slices with a thickness of 0.5 cm, wash once with deionized water, and drain the surface moisture for later use.

[0028] 3.2 Hot water extraction: Add 2.0L of deionized water to the treated chicory root slices at a material-to-liquid ratio of 1:10; place in a constant temperature water bath and stir for 2 hours at 80℃; after extraction, filter through a 100-mesh filter cloth and collect the filtered liquid to obtain 1.8L of extract.

[0029] 3.3 Concentration and Drying: The extract was transferred to a rotary evaporator and concentrated to a solid content of 25wt% at 65℃ and -0.08MPa. The concentrate was then sent to a spray dryer with an inlet air temperature of 180℃ and an outlet air temperature of 85℃. After drying, a light brown powder was collected to obtain 32g of chicory extract.

[0030] 3.4 Quality Inspection: The inulin content was determined by the 3,5-dinitrosalicylic acid (DNS) method, and the result was 63.7±1.2wt%, which meets the requirement of ≥60wt%. The moisture content was determined by the Karl Fischer method, and the moisture content was 4.2wt%, which meets the quality standard of ≤5wt%.

[0031] 4. Preparation process of mulberry leaf extract and hawthorn extract 4.1 Preparation of Mulberry Leaf Extract: Take 0.2 kg of dried mulberry leaves, with an initial total flavonoid content ≥ 5 wt%; pulverize the dried mulberry leaves and pass them through an 80-mesh sieve. Add 2.4 L of 50% ethanol at a material-to-liquid ratio of 1:12, place in a reflux extraction apparatus, and reflux extract twice at 75℃, each extraction lasting 2 hours. After extraction, combine the two filtrates to obtain a total filtrate of 3.8 L; concentrate the filtrate to a solid content of 32 wt%, and then freeze-dry to finally obtain 28 g of green mulberry leaf extract powder.

[0032] 4.2 Preparation of Hawthorn Extract: Take 0.2 kg of pitted dried hawthorn, with an initial triterpenic acid content ≥3 wt%; pulverize the pitted dried hawthorn and pass it through a 60-mesh sieve; use the same extraction process as for mulberry leaf extract: add 50% ethanol at a material-to-liquid ratio of 1:12, and reflux extract twice at 75℃, 2 hours each time. Combine the filtrates after extraction, concentrate to a solid content of 30 wt%, and freeze-dry to obtain 35 g of brownish-red hawthorn extract powder.

[0033] 4.3 Quality Inspection (1) Detection of mulberry leaf extract: The total flavonoid content was determined by aluminum salt colorimetric method. Rutin was used as standard. The total flavonoid content was 32.4±0.8wt%, which met the requirement of ≥30wt%.

[0034] (2) Hawthorn extract detection: The content of triterpenic acid was determined by vanillin-glacial acetic acid colorimetric method. With ursolic acid as standard, the content of triterpenic acid was 16.8±0.5wt%, which meets the quality standard of ≥15wt%.

[0035] 5. Composite core material compounding Weigh out 32g of peppermint extract, 21g of green tea extract, 12g of chicory extract, 9g of mulberry leaf extract, and 16g of hawthorn extract in a mass ratio of 3.2:2.1:1.2:0.9:1.6. Add them to a double cone mixer (SYH-5 type) and stir at 150r / min for 30min in a constant temperature and humidity environment of 25℃ and 45% relative humidity. Store in a sealed container at 4℃ to obtain 100g of composite core material.

[0036] Preparation of a multifunctional palatability-enhancing base material (synergistic effect of quaternary components) (1) Freshwater fish enzymatic hydrolysate - raw material pretreatment: Take 500g of fresh grass carp and 200g of crucian carp, with a mass ratio of 2.5:1; process the fish by removing the scales and internal organs, then wash them three times with deionized water and drain the surface water; cut the processed fish into 1.5cm pieces. 3 The fish pieces were cut into uniform small pieces, resulting in 680g of fish chunks.

[0037] (2) Stepwise enzymatic hydrolysis process: a. First stage enzymatic hydrolysis: Add fish pieces to 3.5L of deionized water at a material-to-liquid ratio of 1:5, heat in an 88℃ water bath for 28min to sterilize; cool naturally to 52℃. Adjust the pH to 7.5 with 0.1mol / L sodium hydroxide; add 0.8g of 25000U / g animal protease (containing 15wt% Bacillus subtilis protease), place in a THZ-82 constant temperature water bath shaker, and stir at 120r / min for 2h for enzymatic hydrolysis.

[0038] b. Second stage of enzymatic hydrolysis: Adjust the pH to 8.4 with 0.1 mol / L sodium hydroxide; cool to 38℃; add 0.8 g 20000 U / g trypsin, and continue enzymatic hydrolysis for 2.3 h. c. Enzyme inactivation and separation: Heat the hydrolyzed material to 95℃ and incubate for 20 min to inactivate the enzyme; then transfer the material to a TGL-16M high-speed centrifuge and centrifuge at 4000 r / min for 13 min, finally taking 3.2 L of supernatant.

[0039] (3) Concentration and solidification: The supernatant was placed in a rotary evaporator and concentrated by rotary evaporation at 65℃ and -0.08MPa until the solid content was 30wt%, yielding 250g of light brown fish hydrolysate.

[0040] (4) Quality testing: The degree of protein hydrolysis was tested using the 2,4,6-trinitrobenzenesulfonic acid (TNBS) method, and the result was 51.3±0.8%, while the index requirement was ≥48%; the free amino acid content was tested using an L-8900 automatic amino acid analyzer, and the result was 18.7±0.6 mg / g.

[0041] Preparation and compounding of three other components 1. Preparation of brewer's yeast extract: 87.5g of high nucleotide type brewer's yeast extract was selected and detected by high performance liquid chromatography (HPLC). The detection conditions were C18 column and mobile phase potassium dihydrogen phosphate buffer-methanol = 95:5. The total content of 5'-inosinate disodium (IMP) and 5'-guanylate disodium (GMP) was found to be 4.6±0.2wt%, which meets the requirement of ≥4wt%.

[0042] 2. Preparation of compound amino acids: Raw materials were weighed at a mass ratio of 2.5:3.5:1.2; the raw materials were 12.5g of taurine (purity 98.5wt%, meeting the ≥98wt% requirement), 17.5g of food-grade L-glycine, and 6.5g of glutamic acid. After the three raw materials were mixed evenly, the total purity was detected by HPLC and found to be 96.2wt%, meeting the ≥95wt% standard, finally yielding 36.5g of compound amino acids.

[0043] 3. Preparation of natural attractants: Take 3.5g of salmon hydrolyzed peptides and analyze them by gel permeation chromatography (GPC). The molecular weight range is 500-1000 Da and the protein content is 90.5 wt%.

[0044] 4. Compounding process: Add 250g of freshwater fish enzymatic hydrolysate, 87.5g of brewer's yeast extract, 36.5g of compound amino acids, and 3.5g of salmon hydrolyzed peptides to a JRJ-10L constant temperature mixing tank. Set the temperature to 50℃ and the stirring speed to 100r / min, and continue stirring for 20min. After stirring, cool to room temperature. The initial mixture is 377.5g. Add deionized water to adjust to 350g, which is the multifunctional palatability-enhancing base material.

[0045] Preparation of four functional excipients (five-element stable components) Weigh each excipient according to the mass ratio of maltodextrin: cryoprotectant: Lactobacillus plantarum: sodium dihydrogen phosphate: tert-butylhydroquinone (TBHQ) = 12:5:1:1.5:0.03, and each excipient must meet the following requirements: 1. Maltodextrin (120g): The DE value was 15 when tested by Fehling's reagent method; the moisture content was 4.8wt% when tested by Karl Fischer method, and the requirement was ≤5wt%; the solubility was 98% at 25℃ when tested according to GB / T5009.3-2016 standard.

[0046] 2. Low-temperature protectant (50g): It is composed of 33.3g of food-grade trehalose and 16.7g of food-grade mannitol in a mass ratio of 2:1, and both have a purity of 99wt%. Through comparative experiments, it was verified that after adding this low-temperature protectant, the survival rate of Lactobacillus plantarum in spray drying can be increased to 1.85 times that without the addition, which meets the requirement of ≥1.8 times.

[0047] 3. *Lactobacillus plantarum* (1g): After being treated at 60℃ for 72 hours, the viability was determined using the plate count method according to GB4789.35-2016 standard. The survival rate was 90.5%, meeting the requirement of ≥90%; the viable count was 1.2 × 10⁻⁶. 9 CFU / g, requirement ≥1×10 9 CFU / g.

[0048] 4. Sodium dihydrogen phosphate (15g): The purity was 99.2wt% by titration, with a requirement of ≥99%; after being prepared into a 1% aqueous solution, the pH value was 6.2; the accelerated oxidation test verified that this component can reduce the oxidation rate of the composite core material by 42%, with a requirement of ≥40%.

[0049] 5. tert-Butylhydroquinone (TBHQ, 0.3g): The purity was 99.5wt% as determined by HPLC; the amount added was 0.03wt% of the total mass of the seasoning powder.

[0050] V. Microcapsule Construction Process (Double-Layer Encapsulation System) 5.1 Construction of inner layer nanocarriers (active ingredient loading) (1) Preparation of L-carnosine liposomes 1. Mixing raw materials: Weigh 17g of L-carnosine (99.5wt% purity) and 17g of dipalmitoylphosphatidylcholine (DPPC) (99wt% purity) at a mass ratio of 1:1, add 20mL of 0.01mol / L, pH7.4 PBS buffer, and stir magnetically for 10min until well mixed.

[0051] 2. Ultrasonic treatment: Place the mixture in a JY92-IIN ultrasonic disruptor and use 200W intermittent ultrasonic treatment for 5 minutes. The ultrasonic mode is 3 seconds on and 2 seconds off. The temperature is controlled at ≤25℃ in an ice bath throughout the process to avoid local overheating and damage to the liposome structure.

[0052] 3. Film formation and hydration: The sonicated liquid was transferred to a RE-52AA rotary evaporator and evaporated at 35℃ and -0.08MPa to form a film to remove the solvent; then 10mL of PBS buffer was added for hydration for 30min to obtain 34g of L-carnosine liposomes.

[0053] 4. Quality Inspection: The particle size was measured using a Zetasizer NanoZS90 Malvern particle size analyzer, with a particle size of 85±12nm; the polydispersity index (PDI) was 0.21, ≤0.3, indicating good dispersibility; and the encapsulation rate was 95.2±0.7% as determined by HPLC, meeting the requirement of ≥95%.

[0054] (2) Preparation of sodium alginate-chitosan composite microspheres 1. Solution preparation: Prepare a 2% (w / v) sodium alginate solution by dissolving it in deionized water and stirring at 150 r / min for 30 min until there are no lumps; prepare a 1% (w / v) chitosan solution by dissolving it in 1% acetic acid and adjusting the pH to 5.5 with 0.1 mol / L sodium hydroxide.

[0055] 2. Mixing and curing: Mix the two solutions at a mass ratio of 4:1; add 0.8wt% CaCl2 solution dropwise at a rate of 2mL / min; and perform electrostatic adsorption curing by stirring at 30℃ and 80r / min for 35min to finally obtain 80g of composite microspheres.

[0056] 3. Quality inspection: The microspheres were measured to be 800 nm using a laser particle size analyzer; and observed to be uniform in morphology and without adhesion by scanning electron microscopy (SEM).

[0057] (3) Preparation of nanocarrier-active ingredient complex: 100g of composite core material and 34g of L-carnosine liposome were added to the composite microsphere suspension at a mass ratio of 1:0.34; the mixture was placed in a constant temperature water bath stirrer and stirred for 35min at 38℃ with a stirring speed of 120r / min to ensure that the active ingredient was fully adsorbed, and finally 114g of the complex was obtained.

[0058] 5.2 Preparation of outer microcapsule wall material (gastric environmental protection) 1. Raw material mixing: Weigh the raw materials according to the mass ratio of gum arabic: gelatin: glycerol: β-cyclodextrin = 3.5: 2.5: 1.2: 0.8. Specifically, weigh 93.1g of gum arabic, 65g of gelatin, 31.92g of glycerol and 76g of β-cyclodextrin.

[0059] 2. Dissolution process: Add the weighed gum arabic, gelatin and glycerin to 200mL of deionized water, and stir at 150r / min for 30min at 68℃ until completely dissolved; after the solution cools naturally to 40℃, add β-cyclodextrin and continue stirring for 28min to finally obtain 396mL of wall material composite solution.

[0060] 3. Quality Inspection Solid content: 20±0.5wt% was measured using a rapid moisture analyzer; Gum arabic viscosity: 280 mPa·s was measured using an NDJ-5S digital viscometer for a 10% aqueous solution of gum arabic; Gelatin properties: 250 Bloom was measured using a TA-XTPlus texture analyzer for gel strength; and gelatin ash content was measured at 1.1wt%, meeting the requirement of ≤1.2wt%.

[0061] 5.3 Double-layer embedding and spray drying (setting process) (1) Complex coagulation reaction: 114g of nanocarrier-active ingredient complex was slowly added dropwise to the wall material composite solution at a volume ratio of 1:3. The mixture was stirred at 38℃ and 90r / min for 1.5h. Then, the pH of the system was adjusted to 4.3 with 1mol / L hydrochloric acid and stirred for 45min to finally form a stable microcapsule suspension.

[0062] (2) Spray drying and quality inspection: The above microcapsule suspension was sent into an LPG-5 spray dryer and the process parameters were set as follows: inlet air temperature 205℃, outlet air temperature 88℃, feed rate 20mL / min, atomization pressure 0.3MPa; after drying, 380g of dried microcapsule powder was collected.

[0063] (3) Quality testing of the collected microcapsule powder: The particle size was measured by a laser particle size analyzer and the result was 18.2 μm; the microspheres were observed by scanning electron microscopy and the surface was smooth and undamaged, meeting the requirements for subsequent processing and use.

[0064] 5.4 Mixing of excipients: Add 380g of microcapsule powder, 120g of maltodextrin, 50g of cryoprotectant, 1g of Lactobacillus plantarum, 15g of sodium dihydrogen phosphate, and 0.3g of TBHQ to a double cone mixer and stir at 130r / min (range 120-150r / min) for 20min (range 18-25min) to obtain 1000g of cat microcapsule flavoring powder containing active plant ingredients.

[0065] Basic indicators: Moisture content detected by rapid moisture meter is 4.2±0.3wt% (≤5wt%); solubility at 25℃ is 93.5±1.2% (≥92%, refer to GB / T5009.3-2016); pH is 6.2.

[0066] 6. Product performance testing 6.1 Physical and chemical performance testing (1) Determination of total encapsulation efficiency Sample preparation: Take 0.1g of flavoring powder, add 10mL of deionized water, sonicate at 300W for 10min to dissolve, pass through a 10kDa ultrafiltration membrane (Millipore type), centrifuge at 4000r / min for 15min, and take the permeate as the free active ingredient sample; take another 0.1g of flavoring powder, add 10mL of 70% ethanol, incubate at 80℃ for 30min to destroy and embed, sonicate for 10min, centrifuge and take the supernatant as the total active ingredient sample.

[0067] HPLC detection: Menthol: mobile phase methanol-0.1% phosphoric acid water = 60:40, wavelength 254nm, the encapsulation efficiency was measured to be 96.2±0.8%; L-carnosine: mobile phase acetonitrile-water = 5:95, wavelength 220nm, the encapsulation efficiency was measured to be 94.8±0.7%; total encapsulation efficiency = (menthol encapsulation efficiency × menthol percentage + L-carnosine encapsulation efficiency × L-carnosine percentage) / 2, calculated to be 95.5±0.6% (≥92%).

[0068] (2) Two-stage targeted release test: Franz diffusion cell (effective diffusion area 1.77 cm²) was used. 2 The receiving solution was physiological saline containing 0.5% Tween 80 (stirred at 37°C and 500 rpm). Simulating the feline gastric environment: Adjusting the pH of the receiving solution to 3.0 (range 2.5-3.5), adding 0.5wt% pepsin, the total release rate within 2 hours was 8.7±0.9% (≤10%); Simulating the feline intestinal environment: Adjusting the pH of the receiving solution to 7.5 (range 6.8-7.8), adding 1wt% pancreatin, the total release rate within 4 hours was 93.5±1.2% (≥92%); Release difference = intestinal release rate - gastric release rate = 84.8±1.5% (≥82%), achieving targeted release.

[0069] 6.2 Functional Verification (1) Palatability test (double bowl method): 30 healthy adult Chinese domestic cats (3.5-4.5kg, half male and half female) were selected and pre-adapted for 7 days (feeding only basic cat food); during the 3-day test period, two food bowls were placed at 8:00 every day: experimental group (basic cat food + 2% flavoring powder) and control group (basic cat food + 2% commercially available chicken liver flavoring attractant), and the positions of the food bowls were switched every day.

[0070] Results: The 24-hour preference rate was 94±2% (28 birds preferred the experimental group); the average feed intake of the experimental group was 285g / bird, while that of the control group was 192g / bird, representing an increase of 48±3% (≥45%) in feed intake.

[0071] (2) Gut health test Grouping: 14 cats were randomly divided into two groups (n=7): experimental group (basic cat food + 2% flavoring powder) and control group (basic cat food + 2% maltodextrin), and fed for 4 weeks.

[0072] Detection: 16S rRNA high-throughput sequencing showed that the relative abundance of Bifidobacteria in feces was 12.8±0.7% in the experimental group and 9.5±0.5% in the control group, an increase of 34.7±2% (range of 25-35%); the incidence of soft stool was 0% in the experimental group and 28.6% in the control group.

[0073] (3) Metabolic improvement test Experimental subjects: 10 obese cats (LDL-C ≥ 3.1 mmol / L) were fed low-fat cat food containing 2% flavoring powder for 8 weeks.

[0074] Results: Serum LDL-C decreased from 3.12±0.21mmol / L to 2.41±0.18mmol / L, a decrease of 22.8±1.2% (≥22%); triglycerides decreased from 1.85±0.13mmol / L to 1.52±0.11mmol / L, a decrease of 17.8±1.0%.

[0075] (4) Anti-aging protection test (DPPH method) Groups: control group (0.1 mg / mL L-carnosine), experimental group (0.1 mg / mL L-carnosine + 0.1 mg / mL tea polyphenols).

[0076] Results: The DPPH scavenging rate in the control group was 78.5±1.8% (≥75%), while that in the experimental group was 128.3±2.1%, which was 1.3 times that of the single component, showing a significant synergistic antioxidant effect.

[0077] 6.3 Stability Test: After vacuum packaging, the seasoning powder was stored under different conditions, and the retention rate of active ingredients was tested periodically. Room temperature storage (25℃, 65%RH): After 6 months, the retention rates of menthol, tea polyphenols, L-carnosine, and Lactobacillus plantarum were 89.1±0.8% (≥88%), 85.3±1.0% (≥85%), and 83.2±0.9% (≥82%), respectively. Accelerated test at 60℃: After 30 days, the retention rates of all active ingredients were ≥85%, and the probiotic survival rate was 85.8±1.3% (≥85%), meeting the 12-month shelf-life requirement.

[0078] Example 2: Preparation and Performance Verification of Gut Health Optimized Formula I. Preparation of composite core material (total preparation amount: 1000g) In this embodiment, the total amount of composite core material is 120g (accounting for 12wt% of the total mass of the finished product). The core adjustment is to increase the amount of chicory extract to the upper limit of the process (to enhance the prebiotic function). The preparation process parameters of the other extracts are the same as in Example 1 to ensure compliance with basic performance.

[0079] 1.1 Composite core material compounding scheme: The plant extracts are compounded in a mass ratio of 2.8:2.0:1.5:1.0:1.7, with the following specific weights: Peppermint extract: 28g (preparation process as described in Example 1); Green tea extract: 20g (preparation process as described in Example 1); Chicory extract: 18g (core incremental component, enhancing prebiotic function, preparation process as described in 2.1.2-2.1.4); Mulberry leaf extract: 10g (preparation process as described in Example 1); Hawthorn extract: 17g (preparation process as described in Example 1).

[0080] 1.2 Chicory Extract—Raw Material Pretreatment: 1.2 kg of fresh chicory root was selected (50% more than in Example 1, matching the requirement of 18 g of extract preparation); after washing the surface impurities with deionized water, it was cut into uniform thin slices of 3-5 cm thickness; the slices were placed in a hot air drying oven and dried at 60°C until the moisture content of the material dropped to 8 wt%; after drying, the material was pulverized and passed through a 60-mesh sieve to remove coarse fiber impurities, and the sieve-passing material was collected to obtain 0.95 kg of chicory powder.

[0081] 1.3 Chicory Extract - Hot Water Extraction Process: Add 9.5L of deionized water to 0.95kg of chicory powder at a material-to-liquid ratio of 1:10; place in a constant temperature water bath environment and extract at 85℃ with stirring at 120r / min for 2.5h; after extraction, filter with a 100-mesh filter cloth and collect the filtered liquid to obtain 8.8L of chicory extract.

[0082] 1.4 Chicory Extract—Purification and Concentration: 8.8 L of chicory extract was filtered through a Millipore 10 kDa ultrafiltration membrane to remove macromolecular impurities (such as crude fiber and protein). The ultrafiltration retentate was collected and transferred to a rotary evaporator, where it was concentrated to a solid content of 32 wt% at 65 °C and -0.08 MPa to obtain 68 g of light brown chicory extract powder. 18 g of this powder was accurately weighed for use in the compound core material formulation.

[0083] 1.5 Quality testing of each extract: Peppermint extract: Supercritical CO2 extraction was used, with parameters the same as in Example 1 (pressure 25 MPa, temperature 45℃, time 3 h). The menthol content was measured to be 21.2 ± 0.5 wt%, meeting the requirement of ≥18 wt%. Green tea extract: After water extraction and purification with AB-8 macroporous resin, the tea polyphenol content was measured to be 29.1 ± 0.7 wt%, meeting the requirement of ≥25 wt%. After being mixed with mulberry leaf extract in a certain proportion, the oxidation half-life of total mulberry leaf flavonoids reached 33 h, an extension factor of ≥2.5 times. Mulberry leaf extract: Extracted by reflux with 50% ethanol (solid-liquid ratio 1:12, extraction twice at 75℃, 2 h each time). The total flavonoid content was measured to be 32.6±0.8wt%, meeting the requirement of ≥30wt%. The triterpenic acid content of hawthorn extract was measured to be 16.9±0.5wt%, meeting the requirement of ≥15wt%. The inulin content of chicory extract was measured using high performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD), with acetonitrile-water mobile phase = 75:25, flow rate 1.0mL / min, column temperature 35℃, and the inulin content was measured to be 68.2±1.5wt% (≥60wt%). The moisture content was determined by Karl Fischer method, and the moisture content was 4.5±0.2wt% (≤5wt%), ensuring storage stability.

[0084] 1.6 Composite Core Material Formulation Process 18g of chicory extract, 33.6g of peppermint extract, 24g of green tea extract, 12g of mulberry leaf extract, and 20.4g of hawthorn extract were added to a double cone mixer. The mixture was stirred at 150r / min for 30min under a constant temperature and humidity environment of 25℃ and 45% relative humidity. After stirring, the mixture was sealed and stored at 4℃ to prevent inulin from absorbing moisture and clumping. The final product was 120g of composite core material (of which the total inulin content was 12.28g, which is 25% higher than that of Example 1, thus strengthening the prebiotic base).

[0085] II. Preparation of Multifunctional Appetizing Base Material (total 336g, accounting for 33.6wt% of the total mass of the finished product): It is compounded in a mass ratio of 5.2:2.4:1.8:0.15, and its digestibility is improved by optimizing the enzymatic hydrolysis process to meet the taste needs of cats with sensitive intestines.

[0086] 2.1 Preparation of freshwater fish enzymatic hydrolysate (260g) 2.1.1 Raw material pretreatment and enzymatic hydrolysis process: Take 520g of fresh grass carp and 208g of crucian carp, with a mass ratio of 2.5:1; process the fish by removing scales and internal organs, washing three times with deionized water, draining the surface moisture, and cutting into 1.5cm pieces. 3 The fish pieces are cut into uniform small pieces; the fish pieces are added to deionized water at a ratio of 1:5, heated in an 88°C water bath for 28 minutes to sterilize, and then naturally cooled to 53°C. First stage of enzymatic hydrolysis: Add 0.85g of 25000U / g animal protease (containing 15% subtilis protease), adjust the pH to 7.5, and hydrolyze for 2 hours; Second stage of enzymatic hydrolysis: Add 0.2% papain (to assist in the degradation of large protein molecules), and hydrolyze for 1 hour at 55℃; Third stage of enzymatic hydrolysis: Add 0.85g of 20000U / g trypsin, adjust the temperature to 38℃ and the pH to 8.4, and hydrolyze for 2.3 hours.

[0087] 2.1.2 Post-processing: The enzymatically hydrolyzed material was heated to 95℃ and kept at that temperature for 20 min to inactivate the enzyme; then it was transferred to a high-speed centrifuge and centrifuged at 4000 r / min for 13 min, and the supernatant was collected; the supernatant was placed in a rotary evaporator and concentrated to a solid content of 30 wt%, yielding 260 g of light brown fish enzymatic hydrolysate.

[0088] 2.1.3 Quality Testing: Degree of protein hydrolysis: The 2,4,6-trinitrobenzenesulfonic acid (TNBS) method was used for testing, and the result was 52.8±0.8%, which meets the requirement of ≥48%; Molecular weight distribution: Gel permeation chromatography (GPC) was used for testing, and the proportion of small molecule peptides <1kDa was 52%, which can improve digestion and absorption rate and reduce intestinal burden.

[0089] 2.2 Preparation of other palatability-enhancing components 2.2.1 Brewer's yeast extract (80.6g): A high-nucleotide type brewer's yeast extract was selected and detected by high performance liquid chromatography (HPLC). The detection conditions were: C18 column, mobile phase potassium dihydrogen phosphate buffer-methanol = 95:5. The total content of 5'-inosinate disodium (IMP) and 5'-guanylate disodium (GMP) was found to be 4.4±0.2wt%, which meets the requirement of ≥4wt%. It has a mild umami taste and does not irritate the intestines.

[0090] 2.2.2 Compound amino acids (60.5g): Taurine, food-grade L-glycine, and glutamic acid were weighed in a mass ratio of 3.0:3.5:1.5; among which, the purity of taurine was 98.6wt% (meeting the requirement of ≥98wt%), which has a protective effect on the intestinal mucosa; after the three raw materials were mixed evenly, the total purity was detected by HPLC and found to be 96.5±0.3wt%, which meets the standard of ≥95wt%.

[0091] 2.2.3 Natural palatability enhancer (4.3g): Take 4.3g of salmon hydrolyzed peptides. The molecular weight range is 500-1000Da as determined by gel permeation chromatography (GPC). Avoid adding highly irritating ingredients and make it suitable for cats with sensitive intestines.

[0092] 2.3 Multifunctional attractant base compounding process: 260g of freshwater fish enzymatic hydrolysate, 80.6g of brewer's yeast extract, 60.5g of compound amino acids, and 4.3g of salmon hydrolyzed peptides were added together into a JRJ-10L constant temperature mixing tank; the temperature was set to 50℃ and the stirring speed to 100r / min, and stirring was continued for 20min; after stirring, the mixture was cooled to room temperature to 25℃ to ensure no odor, and finally 336g of multifunctional attractant base was obtained.

[0093] III. Preparation of functional excipients: Fructooligosaccharides were added in a mass ratio of 10:6:2:1.8:0.03:3 to enhance the intestinal health-related components.

[0094] 3.1 Core Optimized Ingredients 3.1.1 Fructooligosaccharides (FOS, 30g): Food-grade fructooligosaccharides with a purity ≥95wt% and a degree of polymerization of 3-5 are selected, which are easily utilized by bifidobacteria; they form a "dual prebiotic synergistic" effect with inulin in the composite core material, thereby enhancing the intestinal prebiotic function.

[0095] 3.1.2 *Lactobacillus plantarum* (2g): A heat-resistant strain (batch number 20240615) produced by Keto Biotechnology was selected. After treatment at 60℃ for 72 hours, the viability was detected using the plate count method according to GB4789.35-2016 standard. The viability rate was 91.5±0.7% (meeting the requirement of ≥90%). The viable count was 2.1×10⁻⁶. 9 CFU / g (satisfying ≥1×10) 9 The CFU / g index requirement is doubled compared to Example 1, thus enhancing intestinal colonization ability.

[0096] 3.2 Preparation of basic auxiliary materials 3.2.1 Maltodextrin (100g): The DE value was 15 as determined by Fehling's reagent method; the moisture content was 4.7±0.2wt% as determined by Karl Fischer method (meeting the requirement of ≤5wt%); the solubility was 98% at 25℃ as determined by GB / T5009.3-2016 standard.

[0097] 3.2.2 Low temperature protectant (60g): It is a compound of food-grade trehalose and food-grade mannitol in a mass ratio of 3:1, and both have a purity of 99wt%. Through comparative experiments, it was verified that after adding this low temperature protectant, the survival rate of Lactobacillus plantarum in spray drying can be increased to 1.85 times that without the addition (meeting the index requirement of ≥1.8 times).

[0098] 3.2.3 Sodium dihydrogen phosphate (18g): The purity was determined by titration to be 99.3±0.1wt% (meeting the requirement of ≥99%); the pH value was 6.3 after being prepared into a 1% aqueous solution; the accelerated oxidation test verified that this component can reduce the oxidation rate of the composite core material by 42.5% (meeting the requirement of ≥40%).

[0099] 3.2.4 Tert-butylhydroquinone (TBHQ, 0.3g): The purity was 99.5wt% as determined by HPLC; its addition amount was 0.03wt% of the total mass of the seasoning powder, which could inhibit the oxidation of unsaturated fatty acids in fish hydrolysate.

[0100] 3.3 Functional excipient compounding process: First, add 100g of maltodextrin, 60g of cryoprotectant, 18g of sodium dihydrogen phosphate, and 0.3g of TBHQ to a double cone mixer and stir at 120r / min for 15min until uniformly mixed; then add 30g of fructooligosaccharide and 2g of Lactobacillus plantarum, and continue stirring at 100r / min for 10min to avoid premature exposure of probiotics to high temperatures, which could lead to inactivation. Finally, 204g of functional excipients are obtained.

[0101] IV. Microcapsule Construction Process 4.1 Construction of inner layer nanocarrier (total 144g, adapted for intestinal component loading) 4.1.1 Preparation of L-carnosine liposomes (36g) Raw material mixing: Weigh 18g of L-carnosine (99.5wt% purity) and 18g of dipalmitoylphosphatidylcholine (DPPC) (99wt% purity) at a mass ratio of 1:1, add 20mL of 0.01mol / L, pH7.4 PBS buffer, add 0.5% cholesterol (to enhance liposome membrane stability and reduce the damage of the intestinal environment to liposomes), and stir magnetically for 10min until well mixed; Ultrasonic treatment: The mixture was placed in a JY92-IIN ultrasonic disruptor and subjected to 200W intermittent ultrasonic treatment for 5 minutes. The ultrasonic mode was 3 seconds on and 2 seconds off. The temperature was controlled at ≤25℃ in an ice bath throughout the process to avoid local overheating and damage to the liposome structure. Film formation and hydration: The sonicated liquid was transferred to a RE-52AA rotary evaporator and rotary evaporated at 35℃ and -0.08MPa to form a film to remove the solvent; then 10mL of PBS buffer was added for hydration for 30min to obtain 36g of L-carnosine liposomes. Quality testing: The particle size was measured using a Zetasizer NanoZS90 Malvern particle size analyzer. The particle size was 90±10nm, and the polydispersity index (PDI) was 0.22 (≤0.3, indicating good dispersibility). The encapsulation efficiency was 96.8±0.5% by HPLC (meeting the requirement of ≥95%). The stability of L-carnosine was improved by 15% compared with Example 1.

[0102] 4.1.2 Preparation of sodium alginate-chitosan composite microspheres (108g) Solution preparation: Prepare 90 mL of 2% (w / v) sodium alginate solution, dissolve in deionized water, and stir at 150 rpm for 30 min until no lumps form; prepare 30 mL of 1% (w / v) chitosan solution, dissolve in 1% acetic acid, and adjust the pH to 5.5 with 0.1 mol / L sodium hydroxide. Mixing and curing: Mix the two solutions at a mass ratio of 3:1, and add 1.0 wt% CaCl2 solution dropwise at a rate of 2 mL / min; stir at 30℃ and 80 rpm for 35 min for electrostatic adsorption curing, finally obtaining 108 g of composite microspheres. Quality testing: The microsphere particle size was measured to be 820 ± 50 nm using a laser particle size analyzer; scanning electron microscopy (SEM) showed that the microspheres were uniform in morphology and free from aggregation.

[0103] 4.1.3 Preparation of nanocarrier-active ingredient complex: 120g of composite core material and 36g of L-carnosine liposomes were added to the composite microsphere suspension at a mass ratio of 1:0.3; the mixture was placed in a constant temperature water bath stirrer and magnetically stirred at 200r / min for 40min at 37℃ (extending the stirring time to ensure sufficient prebiotic loading); after stirring, the material was collected to obtain 144g of nanocarrier-active ingredient complex; the inulin encapsulation rate was 95.2±0.6% as detected by HPLC-ELSD.

[0104] 4.2 Preparation of outer microcapsule wall material (264g) 4.2.1 Raw material ratio and weighing: Weigh each raw material according to the mass ratio of gum arabic: gelatin: glycerol: β-cyclodextrin = 3.2: 2.4: 1.5: 0.9; wherein the viscosity of gum arabic is 275 mPa・s, the gel strength of gelatin is 245 Bloom, the purity of glycerol is 99.6%, and the moisture content of β-cyclodextrin is 9.8%.

[0105] 4.2.2 Dissolution process: Add the weighed gum arabic, gelatin, and glycerin to deionized water and stir at 150 r / min for 30 min at 68℃ until completely dissolved; after the solution cools naturally to 40℃, add β-cyclodextrin and continue stirring for 28 min; after stirring, the solid content is measured to be 21±0.5wt%, and the final wall material composite solution is 264g.

[0106] V. Microcapsule Construction Process 5.1 Double-layer encapsulation and spray drying (384g of finished microcapsule powder): The microcapsule suspension formed by the coagulation reaction was fed into an LPG-5 spray dryer. To protect the activity of probiotics, nitrogen protection was used throughout the process to reduce oxidative inactivation of probiotics. The spray process parameters were set as follows: Inlet air temperature: 198±2℃, process operating range 190-210℃; outlet air temperature: 86±1℃, process operating range 85-92℃; atomization pressure: 0.28±0.02MPa, process operating range 0.25-0.35MPa; feed rate: 19±1mL / min, process operating range 18-22mL / min.

[0107] After spray drying, 384g of microcapsule powder was collected and subjected to quality testing. Particle size detection: The particle size was measured by a laser particle size analyzer and was 16±2μm, which is within the process range of 10-25μm. Morphology detection: The microspheres were observed to be smooth and free of probiotic leakage by scanning electron microscopy, which meets the requirements for subsequent processing and use.

[0108] 5.2 Mixing of excipients (1000g finished product): Add 384g of microcapsule powder and 204g of functional excipients to a double cone mixer, set the stirring speed to 130r / min, and continue stirring for 20min; after mixing, test the mixing uniformity. The relative standard deviation (RSD) is 1.1%, which meets the uniformity requirement of ≤3%, and finally obtain 1000g of intestinal health optimized cat microcapsule flavoring powder.

[0109] Basic performance tests were conducted on the finished seasoning powder: Moisture content: The result was 4.4±0.3wt% using a rapid moisture analyzer, which meets the quality requirement of ≤5wt%. Solubility: The solubility was tested according to GB / T5009.3-2016 standard. The solubility at 25℃ was 92.8±1.2%, which meets the standard of ≥92%.

[0110] VI. Product Performance Testing 6.1 Physical and chemical performance testing 6.1.1 Determination of Total Encapsulation Efficiency The encapsulation efficiency of inulin and L-carnosine in the composite core material was measured separately, and the total encapsulation efficiency was calculated to verify the loading effect of active ingredients. Inulin encapsulation efficiency: Detected using high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) with acetonitrile-water = 75:25 as the mobile phase, the encapsulation efficiency was 95.2±0.6%. L-Carnosine encapsulation efficiency: Detected using high-performance liquid chromatography (HPLC) with acetonitrile-water = 5:95 as the mobile phase, the encapsulation efficiency was 96.8±0.5%. Total encapsulation efficiency: Calculated as “(inulin encapsulation efficiency × inulin percentage + L-carnosine encapsulation efficiency × L-carnosine percentage) / 2”, the result was 96.0±0.6%, meeting the requirement of ≥92%, indicating excellent prebiotic and probiotic loading effects.

[0111] 6.1.2 Two-stage targeted release test: The Franz diffusion cell method was used (effective diffusion area 1.77 cm²).2 Using physiological saline containing 0.5% Tween 80 as the receiving solution (at a constant temperature of 37°C and with stirring at 500 rpm), the targeted release performance was verified by simulating the feline gastric and intestinal environment. Simulated gastric environment test: The pH of the receiving solution was adjusted to 3.0, and 0.5 wt% pepsin was added. The total release rate of active ingredients within 2 hours was 7.2 ± 0.6%, meeting the requirement of ≤10%. Inulin and probiotics were hardly released, which can avoid destruction by gastric acid. Simulated intestinal environment test: The pH of the receiving solution was adjusted to 7.5, and 1 wt% pancreatin was added. The total release rate of active ingredients within 4 hours was 94.3 ± 1.1%, meeting the requirement of ≥92%. Prebiotics and probiotics can be released simultaneously. Release difference: The difference between the intestinal release rate and the gastric release rate was 87.1 ± 1.3%, meeting the requirement of ≥82%, and the targeting is better than that of Example 1.

[0112] 6.2 Validation of Gut Health Function 6.2.1 In vitro gut microbiota culture test The regulatory effect of seasoning powder on gut microbiota was verified by in vitro culture, and the process was designed as follows: 1. Fecal sample preparation: Take 10g of fresh feces from a healthy adult cat, dilute it with sterile saline to a suitable concentration (to ensure bacterial activity), and remove impurities through sterile filtration; 2. Culture medium preparation: Add the diluted fecal bacterial solution to a sterile culture medium containing this flavoring powder, so that the final concentration of the flavoring powder is 2%; 3. Culture conditions: Place the above culture medium in a 37℃ anaerobic incubator and culture for 48 hours (simulating the anaerobic environment of the intestine). 4. Detection Indicators: After cultivation, the number of Bifidobacteria in the culture medium was determined by plate counting, and the content of short-chain fatty acids (SCFAs, such as propionic acid and butyric acid) in the culture medium was determined by gas chromatography to evaluate the promoting effect of the flavoring powder on the proliferation of beneficial bacteria and the production of short-chain fatty acids. The results are as follows:

[0113] 6.2.2 In vivo gut health validation test (gut-sensitive cat model) 1. Selection and grouping of test subjects Fifteen clinically diagnosed cats with gut sensitivities were selected as subjects for the experiment. The specific selection criteria and grouping are as follows: Basic conditions: weight 3.0-4.0kg, no other systemic diseases, to ensure that the test results only reflect changes in intestinal status; Sensitivity determination: loose stools or diarrhea ≥3 times per week, which meets the clinical characteristics of intestinal sensitivity; Randomization: cats were randomly divided into two groups using a random number table method, with 7 cats in the experimental group and 8 cats in the control group. There were no significant differences in weight and initial intestinal sensitivity between the two groups (P>0.05), ensuring comparability between the groups.

[0114] 2. Test Treatment Plan Both groups were fed the same brand and formula of basic cat food as their staple diet, with the only difference being the added ingredients. The specific treatments are as follows: Experimental group: Basic cat food + 3% of the "Gut Health Optimized Cat Microcapsule Flavoring Powder" prepared in this embodiment, fed daily with the cat food; Control group: Basic cat food + 3% of commercially available probiotic powder (this probiotic powder contains only a single strain and does not contain inulin, fructooligosaccharides, or other prebiotic ingredients), fed in the same way as the experimental group to exclude the interference of "simple probiotic addition" on the experimental results.

[0115] 3. Experimental period and observation indicators Experiment period: lasting 6 weeks, during which the rearing environment (temperature, humidity, cage space), feeding time, and daily feed amount were kept consistent to avoid the influence of additional variables; Observation indicators: Record the following indicators at fixed times each week: 1. Fecal score: A 1-5 point scoring system was used, with 5 points being the best (formed stool, moderate texture) and 1 point being the worst (watery diarrhea). The average fecal score of each group of cats was recorded. 2. Diarrhea frequency: The total number of times each group of cats had soft stools or diarrhea per week was counted, and the average weekly diarrhea frequency of a single cat was calculated. 3. Hair quality: Visual observation and tactile assessment (hair smoothness, shine) were used to help determine whether the overall hair health had improved. Results are as follows:

[0116] 6.3 Stability Test 6.3.1 Stability Test During Room Temperature Storage: The gut health-optimized seasoning powder of this embodiment was vacuum-packed and stored at 25℃ and 65% relative humidity for 6 months. Samples were periodically taken to test the retention rate of the core functional components: Inulin retention rate: 88.5±0.8%, meeting the stability requirement of ≥85%; Lactobacillus plantarum survival rate: 86.2±1.2%, meeting the stability requirement of ≥85%; Bifidobacterium proliferation capacity: After 48 hours of in vitro culture, the number of Bifidobacterium still reached 9.2×10⁻⁶. 8 The CFU / g value decreased by only 6.1% from the initial value, indicating that its function of promoting the proliferation of beneficial intestinal bacteria was not significantly diminished.

[0117] 6.3.2 Accelerated Stability Test To further verify long-term storage reliability, two accelerated aging conditions were set up for testing: 1. High temperature and high humidity acceleration: The seasoning powder was stored in an environment of 40℃ and 75% relative humidity for 3 months. The results showed that the inulin retention rate was 84.2±1.0% and the survival rate of Lactobacillus plantarum was 83.7±1.3%.

[0118] 2. High temperature acceleration: The seasoning powder was stored in an environment of 60℃ for 30 days. The results showed that the inulin retention rate was 82.5±1.1% and the survival rate of Lactobacillus plantarum was 81.9±1.2%.

[0119] Both of the above accelerated test results meet the stability requirements, ensuring that intestinal function is effectively preserved during the product's 12-month shelf life.

[0120] 6.4 Palatability Test 6.4.1 Experimental Design Subjects: Thirty healthy adult cats were selected, including eight cats with sensitive gut (consistent with the screening criteria for cats with sensitive gut in experiment 5.2.2) to ensure that the experimental results are relevant to the target user group; Experimental method: A double-bowl comparison method was used, with two food bowls of equal quantity placed in the cat house at 8:00 AM daily. Experimental group food bowl: basic cat food + 3% of the gut health-optimized flavoring powder in this embodiment; Control group food bowl: basic cat food + 3% commercially available chicken liver flavored palatability enhancer; Experiment duration: 3 days, with the positions of the two food bowls being switched daily to avoid interference from environmental location on the cat's food selection.

[0121] 6.4.2 Test Results Feed preference rate: Within 3 days, 27 cats preferred the experimental group's food bowl, with a preference rate of 91±2%, meeting the palatability index requirement of ≥90%; Feed intake comparison: The average daily feed intake per cat in the experimental group was 278±10g, while that in the control group was 205±8g, with the experimental group's feed intake increasing by 35.6% compared to the control group; Applicability statement: Although this palatability index is slightly lower than that of Example 1 (standard formula), it is sufficient to ensure the basic feeding willingness of cats with sensitive intestines and avoid insufficient intake of functional components due to poor palatability.

[0122] In summary, to ensure the quality stability and functional effectiveness of the industrialized production of gut health-optimized seasoning powder, three core aspects need to be carefully controlled: First, standardized management of chicory extract. Each batch must be tested for inulin content using HPLC-ELSD to ensure it is ≥60wt%. Simultaneously, the storage environment must be controlled at a temperature ≤25℃ and relative humidity ≤40% to prevent the extract from absorbing moisture and clumping, which could affect subsequent compounding. Second, ensuring the activity of probiotics throughout the entire process. Before spray drying, the suspension temperature must be controlled at ≤40℃ (as the survival rate of probiotics drops sharply above 45℃), and the finished product should be vacuum-packed in aluminum foil composite bags to ensure an oxygen permeability of <0.5cc / m³.2 The product is kept away from light to reduce the damage to probiotic activity caused by oxygen and light. Thirdly, the synergistic ratio of prebiotics is controlled. Inulin and fructooligosaccharides need to be compounded at a mass ratio of 1:1.67. This ratio has been verified by experiments to achieve the best effect of bifidobacteria proliferation, while avoiding bloating in cats caused by excessive ratio, thus ensuring a balance between product function and palatability.

[0123] Comparative Example 1: Single-layer embedding system 1. Raw material composition and proportions: The total composition of the raw materials in this comparative example is consistent with that of the "Cat Microcapsule Flavoring Powder Containing Active Plant Ingredients" in this application. The core difference is that only a single-layer encapsulation system is used (without an inner nano-carrier). The specific proportions (based on 100% of the total mass) and key components are described below: 1.1 Mass percentage and details of each component Composite core material (12%): a compound of peppermint extract, green tea extract, chicory extract, mulberry leaf extract, and hawthorn extract in a mass ratio of 3:2:1.2:0.8:1.5; Multifunctional attractant base (35%): a compound of freshwater fish enzymatic hydrolysate, brewer's yeast extract, compound amino acids, and natural attractants in a mass ratio of 5:2:1.5:0.1; Single-layer encapsulation system (40%): only a mixture of gum arabic and gelatin (mass ratio 3.5:2.5, consistent with the ratio of the two components in the outer wall material of the product of this application), without the inner nanocarrier (sodium alginate-chitosan complex + liposome-encapsulated L-carnosine) of the product of this application, and without glycerol and β-cyclodextrin in the outer wall material of the product of this application; Functional excipients (13%): maltodextrin, cryoprotectant, thermoresistant probiotics, sodium dihydrogen phosphate, and tert-butylhydroquinone are compounded in a mass ratio of 10:4:1:1.98:0.02; the cryoprotectant is composed of trehalose and mannitol in a mass ratio of 2:1, and the thermoresistant probiotics are Lactobacillus plantarum (live count ≥1×10⁻⁶). 9 CFU / g).

[0124] 2. Preparation method 2.1 Preparation of basic components: Preparation of composite core material: Peppermint extract was prepared by supercritical CO2 extraction (extraction pressure 26MPa, temperature 45℃, time 3h), and the preparation process of other extracts (green tea, chicory, mulberry leaf, hawthorn) was the same as that in this application. Preparation of multifunctional attractant base: Freshwater fish enzymatic hydrolysate is prepared using a two-step enzymatic hydrolysis process (first adding animal protease and hydrolyzing at 53℃ for 2 hours, then adding trypsin and hydrolyzing at 38℃ for 2.2 hours). The preparation process of brewer's yeast extract, compound amino acids, and natural attractants is consistent with that of this application. Preparation of functional excipients: The purity testing and pretreatment of maltodextrin, sodium dihydrogen phosphate, and tert-butylhydroquinone, as well as the compounding process of the cryoprotectant, are consistent with those in this application.

[0125] 2.2 Single-layer embedding process 2.2.1 Mixing of active ingredients: The composite core material and L-carnosine are mixed at a mass ratio of 1:0.4, without encapsulation of the inner nanocarrier (sodium alginate-chitosan complex + liposomes) of the product of this application; 2.2.2 Wall material dissolution: Mix gum arabic and gelatin at a mass ratio of 3.5:2.5, add deionized water and dissolve at 68°C, adjust the solid content to 20wt% to obtain a single-layer embedded wall material solution; 2.2.3 Complex coagulation reaction: The "composite core material-L-carnosine mixture" was slowly added dropwise to the monolayer embedded wall material solution at a volume ratio of 1:3, and stirred for 1.5 h at 40 °C and 90 r / min. Then, the pH of the system was adjusted to 4.4 with acid, and stirring was continued for 45 min to form a stable microcapsule suspension.

[0126] 2.3 Spray drying and mixing with finished product 2.3.1 Spray drying: The microcapsule suspension was fed into a spray dryer, and the process parameters were set as follows: inlet air temperature 200℃, outlet air temperature 88℃, feed rate 20mL / min, and atomization pressure 0.3MPa. After drying, the microcapsule powder was collected. 2.3.2 Mixing of excipients: Add the microcapsule powder and functional excipients together to the mixing equipment and stir at 140 r / min for 20 min. After mixing evenly, the final product of Comparative Example 1 is obtained.

[0127] 3. Performance Testing

[0128] 4. Comparative Analysis of Comparative Example 1 and the Product of this Application 4.1 Differences in targeted release performance in the gastric environment: The lack of an inner nanocarrier leads to premature release of active ingredients; the monolayer encapsulation system of Comparative Example 1 did not include the inner nanocarrier (sodium alginate-chitosan pH-sensitive barrier) of the product of this application, which is the core structure for achieving gastric environmental protection - the sodium alginate-chitosan complex can maintain structural stability in a gastric environment with pH≤4.0, avoiding premature degradation of the wall material.

[0129] Due to the absence of this barrier, the single-layer gum arabic-gelatin wall material of Comparative Example 1 is prone to rapid degradation in the gastric environment (pH 2.5-3.5), resulting in the premature release of active ingredients (inulin, L-carnosine, probiotics). The 2-hour release rate in the stomach is significantly higher than that of the product of this application (7.2±0.6%), which fails to achieve the targeted effect of "retention in the stomach and release in the intestines". Consequently, the active ingredients are destroyed by gastric acid, reducing the efficiency of intestinal absorption.

[0130] 4.2 Defects in wall material performance: The lack of key components in the outer layer leads to a decrease in density and thermal stability; the single-layer encapsulation system of Comparative Example 1 did not contain glycerol and β-cyclodextrin, which are crucial components for the performance of the wall material in this application. Glycerin's plasticizing effect can improve the flexibility of the wall material and reduce cracking during drying; β-cyclodextrin's inclusion effect can enhance the density of the wall material and reduce the impact of external environment (such as temperature and oxygen) on the internal active ingredients. Due to the lack of the above components, the single-layer wall material of Comparative Example 1 has insufficient density, is prone to cracking during spray drying (air inlet temperature 200℃), and its thermal stability decreases significantly during storage. The active ingredients (such as probiotics) are deactivated at a faster rate due to temperature.

[0131] 4.3 Overall Functional Differences: Reduced Encapsulation Ratio Confirms the Core Value of the Double-Layer Encapsulation System In summary, the total encapsulation efficiency (inulin + L-carnosine) of the single-layer encapsulation system in Comparative Example 1 was significantly lower than that of the product of this application (96.0±0.6%), and the survival rate of probiotics (stored at room temperature for 6 months) was only 75.3±1.5% (86.2±1.2% of the product of this application).

[0132] The results confirm that the dual-layer encapsulation system of the product (inner nanocarrier for gastric protection + outer wall material for enhanced density and stability) is the key to ensuring the dual functions of "targeted release" and "activity retention", while the single-layer encapsulation system cannot meet the core performance requirements of the product due to structural and component defects.

[0133] Comparative Example 2: Ternary Plant Composite Core Material 1. Raw material composition and proportions The overall composition framework of the raw materials in this comparative example is consistent with that of the "Cat Microcapsule Flavoring Powder Containing Active Plant Ingredients" in this application. The core difference is that the composite core material only retains the ternary plant components (lacking the chicory extract and mulberry leaf extract from the quinary component). The specific proportions (based on 100% of the total mass) and key component descriptions are as follows: 1.1 Mass percentage and details of each component The ternary plant composite core material (12%) is a compound of peppermint extract, green tea extract, and hawthorn extract in a mass ratio of 3:2:1.5, which is completely consistent with the ratio of these three components in the product of this application; the multifunctional palatability-enhancing base material (35%) has the same composition and preparation process as the product of this application, ensuring no difference in palatability and avoiding interference with the comparison of core material functions; the double-layer encapsulation system (40%) has the same composition and preparation process as the product of this application, ensuring consistent encapsulation conditions, and only the difference in core material reflects functional synergy; the functional excipients (13%) have the same composition and ratio as Comparative Example 1 (i.e., maltodextrin: cryoprotectant: thermoresistant probiotics: sodium dihydrogen phosphate: tert-butylhydroquinone = 10:4:1:1.98:0.02, and trehalose: mannitol in the cryoprotectant = 2:1), eliminating the influence of excipient differences on function.

[0134] 1.2 Key Component Description The ternary plant composite core material in this comparative example is clearly missing two key components found in the pentagonal composite core material of this application: chicory extract (in this application, inulin content ≥60wt%, a core prebiotic component); and mulberry leaf extract (in this application, total flavonoid content ≥30wt%, synergistic antioxidant with green tea extract). The preparation processes for the other three extracts (peppermint, green tea, and hawthorn) are the same as those for this application: peppermint extract is extracted using supercritical CO2 extraction, green tea extract is purified using water extraction-AB-8 macroporous resin, and hawthorn extract is extracted using 50% ethanol reflux extraction, ensuring that the single variable is "the number of core material components (ternary vs. pentagonal)".

[0135] 2. Preparation method 2.1 Preparation of ternary plant composite core material: Each extract was weighed according to the mass ratio of "peppermint extract: green tea extract: hawthorn extract = 3:2:1.5" and added to a double cone mixer; in a constant temperature and humidity environment of 25℃ and 45% relative humidity, it was stirred at 150r / min for 30min. After stirring, it was sealed and stored at 4℃ to obtain ternary plant composite core material. The preparation environment and mixing parameters were the same as those of the pentagonal core material of this application.

[0136] 2.2 Double-layer encapsulation process: Inner layer nanocarrier construction: Sodium alginate-chitosan composite microspheres (sodium alginate to chitosan mass ratio 4:1) and L-carnosine liposomes were prepared according to the process of this application. The ternary plant composite core material and L-carnosine liposomes were mixed at a mass ratio of 1:0.34 and added to the microsphere suspension. The mixture was stirred at 38℃ and 120 r / min for 35 min to obtain the nanocarrier-active ingredient complex. Outer layer wall material preparation: A wall material composite solution was prepared according to the ratio of this application (gum arabic:gelatin:glycerol:β-cyclodextrin = 3.5:2.5:1.2:0.8). The dissolution temperature, stirring time, and other parameters were the same as in this application. Complex coagulation reaction: The complex was added dropwise to the outer layer wall material solution at a volume ratio of 1:3. The mixture was stirred at 38℃ and 90 r / min for 1.5 h. After adjusting the pH to 4.3, the mixture was stirred for another 45 min to obtain the microcapsule suspension.

[0137] 2.3 Spray drying and mixing with excipients Spray drying: An LPG-5 spray dryer was used, with the inlet air temperature set at 205℃, the outlet air temperature at 88℃, the feed rate at 20mL / min, and the atomization pressure at 0.3MPa, the same parameters as in this application; Excipient mixing: The spray-dried microcapsule powder and functional excipients were mixed according to the process of this application (stirring at 130r / min for 20min) to obtain the final product of Comparative Example 2.

[0138] Performance testing

[0139] 4. Comparative Analysis of Comparative Example 2 and the Product of this Application 4.1 Weakened intestinal probiotic function: The lack of chicory extract (prebiotic) leads to a decrease in the proliferation capacity of Bifidobacteria; the ternary plant composite core material in this comparative example does not contain chicory extract (core prebiotic source, inulin content ≥60wt%) found in the product of this application, while inulin, as a specific carbon source for Bifidobacteria, can significantly promote its proliferation.

[0140] Comparative tests showed that the number of Bifidobacteria in Comparative Example 2 was 7.5 × 10⁻⁶ after 48 hours of in vitro culture. 8 CFU / g, compared to the product in this application (9.8×10⁻⁶). 8 The CFU / g content decreased by 23.5%, and the production of short-chain fatty acids (propionic acid and butyric acid) decreased by 31.2% and 28.6%, respectively. In in vivo experiments, the improvement rate of soft stool in cats with sensitive intestines was only 57.1%, lower than the 85.7% of the product in this application. This difference indicates that the absence of chicory extract directly weakens the intestinal probiotic function of the core material, confirming its role as the "core support for intestinal health" in the five-element core material.

[0141] 4.2 Decline in Antioxidant and Metabolic Improvement Functions: The absence of mulberry leaf extract (total flavonoids) leads to the loss of synergistic effects. This comparative example of a ternary plant composite core material does not contain mulberry leaf extract (total flavonoid content ≥30wt%) found in the product of this application. Mulberry leaf total flavonoids are a key synergistic antioxidant target of green tea polyphenols—the two can synergistically scavenge free radicals and inhibit lipid peroxidation, thereby enhancing antioxidant and metabolic improvement effects.

[0142] Comparative tests showed that the DPPH free radical scavenging rate of Comparative Example 2 was 92.3±1.5%, a decrease of 28.1% compared to the product of this application (128.3±2.1%); the serum LDL-C reduction in obese cats after 8 weeks of feeding was 15.2±1.0%, a decrease of 33.3% compared to the product of this application (22.8±1.2%). These results indicate that the absence of mulberry leaf extract leads to the loss of synergistic targets for green tea polyphenols, resulting in a significant decline in their antioxidant and metabolic improvement functions, highlighting its "synergistic effect" in the five-element core material.

[0143] 4.3 Decreased palatability: Insufficient flavor profile of the ternary core material leads to reduced willingness to eat. The comparative example of the ternary plant composite core material contains only three extracts: peppermint, green tea, and hawthorn. It lacks the mildly sweet flavor of chicory extract and the crisp flavor of mulberry leaf extract found in the product of this application, resulting in a single flavor profile.

[0144] Palatability testing showed that the cats in Comparative Example 2 had a food preference rate of 78±3%, a decrease of 14.3% compared to the product in this application (91±2%); the average daily food intake per cat was 232±9g, a decrease of 16.5% compared to the product in this application (278±10g); and cats with sensitive intestines exhibited intermittent food refusal due to low flavor acceptance. This difference confirms that the five-element core material achieves "palatability optimization" through the combination of multiple flavor components, while the three-element core material, due to insufficient flavor dimensions, cannot meet the cats' food preference needs.

[0145] 4.4 Advantages of the Five-Element Plant Composite Core Material of this Application: "Functional Synergy + Palatability Optimization" The performance defects of Comparative Example 2 demonstrate that the five-element plant-based composite core material (mint + green tea + chicory + mulberry leaf + hawthorn) of this application is not a simple additive combination of components. Functionally, chicory extract (prebiotic) and Lactobacillus plantarum form a "prebiotic-probiotic synergy," while mulberry leaf extract (total flavonoids) and green tea extract (tea polyphenols) form an "antioxidant synergy," jointly enhancing intestinal health and improving metabolism. In terms of palatability, the complementary flavors of the multiple components (refreshing mint, fragrant green tea, subtly sweet chicory, crisp mulberry leaf, and slightly sour hawthorn) cater to cats' taste preferences. The ternary core material, lacking key synergistic components and flavor dimensions, cannot achieve the above effects, further confirming the rationality and advantages of the five-element core material design of this application.

[0146] Comparative Example 3: Traditional single enzymatic hydrolysis process 1. Raw material composition and proportion: The overall composition framework of the raw materials in this comparative example is completely consistent with that of the "Cat Microcapsule Seasoning Powder Containing Active Plant Ingredients" in this application. The core difference is that the freshwater fish enzymatic hydrolysate in the multifunctional palatability-enhancing base is prepared using a traditional single enzymatic hydrolysis process (this application uses a two-step composite enzymatic hydrolysis process). The specific proportions (based on 100% of the total mass) and key components are described below: 1.1 Mass percentage and details of each component Composite core material (12%): The composition, ratio and preparation process are the same as the product of this application, ensuring that the core material function is the same and avoiding interference with the comparison of enzymatic hydrolysis process; Multifunctional attractant base (35%): A compound of "single-enzyme hydrolysate of freshwater fish," brewer's yeast extract, compound amino acids, and natural attractants, in a mass ratio of 5:2:1.5:0.1. Only the preparation process of the "freshwater fish hydrolysate" differs from this application; the preparation processes of the other three components are the same as in this application. Double-layer encapsulation system (40%): The composition, ratio, and preparation process are the same as the product in this application, eliminating the interference of encapsulation conditions on the final performance. Functional excipients (13%): The composition and ratio are the same as Comparative Example 1 (i.e., maltodextrin: cryoprotectant: thermoresistant probiotic: sodium dihydrogen phosphate: tert-butylhydroquinone = 10:4:1:1.98:0.02, with trehalose:mannitol in the cryoprotectant = 2:1), ensuring no additional variables in the excipients.

[0147] 1.2 Key Differences Explanation The enzyme selection and enzyme activity of this comparative example, "single-enzymatic hydrolysis of freshwater fish hydrolysate," strictly correspond to a portion of the enzyme activity in the two-step complex enzymatic hydrolysis of this application—only 3750 U / g of Bacillus subtilis protease is used. This activity is completely consistent with the activity of Bacillus subtilis protease in the animal protease of this application (the animal protease activity of this application is 25000 U / g, of which 15wt% is Bacillus subtilis protease, corresponding to an activity of 25000 U / g × 15% = 3750 U / g), ensuring that the total enzyme activity is consistent, with only the enzymatic hydrolysis process (single enzyme vs. complex enzyme) being the variable.

[0148] 2. Preparation method 2.1 Preparation of enzymatic hydrolysates from freshwater fish using a single enzyme 2.1.1 Raw material pretreatment: Take fresh grass carp and crucian carp, mix them at a mass ratio of 2.5:1 (consistent with the fish raw material ratio of the two-step enzymatic hydrolysis in this application); remove the scales and internal organs, wash three times with deionized water, drain the surface water, and cut into 1-2cm pieces. 3 Uniform small pieces (consistent with the raw material cutting specifications of this application).

[0149] 2.1.2 Single enzymatic hydrolysis process: Fish pieces are added to deionized water at a material-to-liquid ratio of 1:5, heated in an 88°C water bath for 28 minutes for sterilization (consistent with the sterilization conditions of this application), and then naturally cooled to 53°C (consistent with the first-stage enzymatic hydrolysis temperature of this application); the pH of the system is adjusted to 7.5 (consistent with the first-stage enzymatic hydrolysis pH of this application), and only 3750 U / g of Bacillus subtilis protease is added. Enzymatic hydrolysis is carried out at 120 r / min for 4.2 h (the total enzymatic hydrolysis time is consistent with the total time of the two-step enzymatic hydrolysis of this application, which is "first stage 2 h + second stage 2.2 h").

[0150] 2.1.3 Post-processing: After enzymatic hydrolysis, the material is heated to 95℃ and kept at that temperature for 20 min to inactivate the enzyme (consistent with the enzyme inactivation conditions of this application); then it is transferred to a high-speed centrifuge and centrifuged at 4000 r / min for 14 min (consistent with the centrifugation conditions of this application), and the supernatant is collected; the supernatant is placed in a rotary evaporator and concentrated to a solid content of 30 wt% to obtain single-enzymatic hydrolysate of freshwater fish.

[0151] 2.2 Other preparation steps Composite core material preparation: Mix according to the proportions and processes of the five-element plant composite core material of this application to ensure consistent core material performance; Multifunctional attractant base compound: The single enzymatic hydrolysate of freshwater fish is mixed with brewer's yeast extract, compound amino acids and natural attractants in a certain proportion, and the stirring conditions are the same as those in this application. The double-layer encapsulation process: the construction of the inner nanocarrier (sodium alginate-chitosan composite microspheres + L-carnosine liposomes), the preparation of the outer wall material (gum arabic-gelatin-glycerol-β-cyclodextrin) and the complex coagulation reaction were all carried out according to the parameters of this application; Spray drying and mixing of auxiliary materials: The spray drying parameters (inlet air temperature 205℃, outlet air temperature 88℃, etc.), mixing speed and time of auxiliary materials are completely consistent with those of this application, and the final product of Comparative Example 3 is obtained.

[0152] 3. Performance testing (refer to the product testing methods in this application)

[0153] 4. Comparative Analysis 4.1 Insufficient enzymatic hydrolysis efficiency: Bacillus subtilis protease alone is ineffective at breaking down large fish protein molecules. Comparative Example 3 used a single Bacillus subtilis protease, while this application employs a two-step complex enzymatic hydrolysis method involving "animal protease + trypsin" (animal protease initially hydrolyzes myofibril proteins, and trypsin further hydrolyzes large molecules such as collagen, forming a synergistic effect). Tests showed that Comparative Example 3 had a protein hydrolysis degree of 38.5 ± 0.7% (a decrease of 27.1% compared to this application's 52.8 ± 0.8%), and the proportion of <1kDa small molecule peptides was 32% (significantly lower than this application's 52%). This difference indicates that a single enzyme cannot overcome the degradation barrier of large molecule proteins, resulting in low protein utilization and affecting the subsequent absorption of nutrients from the attractant substrate.

[0154] 4.2 Significantly reduced palatability: Insufficient flavor compounds weaken synergistic flavor enhancement. The palatability of fish enzymatic hydrolysates depends on flavor peptides (such as glutamate peptides) and 5'-nucleotides. Single enzymatic hydrolysis has limitations: Bacillus subtilis protease hydrolysis has narrow specificity, generating only a small amount of glutamate peptides and not promoting the release of 5'-nucleotides. In the two-step enzymatic hydrolysis of this application, animal protease releases basic flavor peptides, and trypsin further generates highly active glutamate peptides and promotes the release of 5'-nucleotides (IMP, GMP), forming a "flavor synergy" with brewer's yeast extract (containing natural flavor substances). Palatability test: The feed preference rate of the three cats in the comparative example was 72±3% (a decrease of 20.9% compared to 91±2% in this application), and the average daily feed intake was 215±8g (a decrease of 22.7% compared to 278±10g in this application), confirming that single enzymatic hydrolysis is difficult to satisfy the feed preferences of cats.

[0155] 4.3 Deterioration in product stability: Low degree of hydrolysis of protein accelerates fatty acid oxidation. Comparative Example 3, with its low degree of hydrolysis (high proportion of macromolecules), readily reacts with unsaturated fatty acids, accelerating oxidation. After 30 days of storage at room temperature, the peroxide value (POV) of Comparative Example 3 was 8.6 ± 0.3 meq / kg (a 65.4% increase compared to 5.2 ± 0.2 meq / kg in this application), exhibiting a rancid taste and a palatability degradation rate of 18% (compared to only 5% in this application). These results indicate that single-step enzymatic hydrolysis of low-hydrolysis proteins affects both palatability and stability; the combined enzymatic hydrolysis in this application, by increasing the degree of hydrolysis and reducing the interaction between protein and fatty acids, balances both aspects. 4.4 Core Conclusion: Two-step combined enzymatic hydrolysis achieves a dual improvement in "protein utilization + flavor." Comparative Example 3 confirms that the two-step process of "animal protease + trypsin" in this application is not a simple superposition of enzymes: In terms of efficiency: the specific complementary nature of enzymatic hydrolysis overcomes the bottleneck of macromolecular protein decomposition, increases the degree of hydrolysis and the proportion of small molecule peptides, and enhances nutrient absorption; in terms of function: it promotes the formation of flavor peptides and 5'-nucleotides, synergistically enhancing flavor with brewer's yeast extract and ensuring palatability; in terms of stability: it reduces the promotion of fatty acid oxidation by low-hydrolyzed proteins, extending shelf life. Single Bacillus subtilis protease has a limited function and cannot achieve the above effects, highlighting the rationality and core value of the process in this application.

[0156] The embodiments of the present invention have been described in detail above, but the content described is only a preferred embodiment of the present invention and should not be considered as limiting the scope of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the patent coverage of the present invention.

Claims

1. A microcapsule flavoring powder for cats containing active plant ingredients, characterized in that, It includes a double-layer encapsulation system, and by weight percentage, the raw material composition is: composite core material 10-14wt%, multifunctional palatability-enhancing base material 32-36wt%, double-layer encapsulation system 35-39wt%, and functional auxiliary materials 14-16wt%. The composite core material is composed of peppermint extract, green tea extract, chicory extract, mulberry leaf extract, and hawthorn extract in a mass ratio of (1.8-4.5):(1.2-3):(0.8-1.5):(0.5-1.2):(1-2). The multifunctional attractant base is composed of freshwater fish enzymatic hydrolysate, brewer's yeast extract, compound amino acids and natural attractants in a mass ratio of (4-7):(1.5-3):(1-2):(0.05-0.15); The bilayer encapsulation system comprises an inner nanocarrier and an outer microcapsule wall material. The inner nanocarrier is a mixture of sodium alginate-chitosan complex and liposome-encapsulated L-carnosine, with a mass ratio of 3-5:1 and a particle size of 50-100 nm. The outer microcapsule wall material is a mixture of gum arabic, gelatin, glycerol, and β-cyclodextrin, with a mass ratio of 2.5-4.5:1.5-3.5:0.8-1.8:0.5-1.2 and a particle size of 10-25 μm. The functional excipients include maltodextrin, a cryoprotectant, thermoresistant probiotics, sodium dihydrogen phosphate, and tert-butylhydroquinone, which are mixed in a mass ratio of (8.5-18):(3-8):(0.5-2):(0.8-2.2):(0.015-0.05). The cryoprotectant is composed of trehalose and mannitol in a mass ratio of 1:1-3:

1.

2. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, Each extract in the composite core material meets the following quality indicators: The menthol content in the peppermint extract is ≥18 wt%; The tea polyphenol content in green tea extract is ≥25wt%; The inulin content in chicory extract is ≥60wt%; The total flavonoid content in mulberry leaf extract is ≥30wt%; The triterpenic acid content in hawthorn extract is ≥15wt%.

3. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, The multifunctional palatability-enhancing base material includes: The degree of protein hydrolysis of freshwater fish enzymatic hydrolysate is ≥48%, and the solid content is 28-32 wt%. The total content of disodium 5'-inosinate and disodium 5'-guanylate in brewer's yeast extract is ≥4 wt%; The compound amino acid is composed of taurine, food-grade L-glycine, and glutamic acid in a mass ratio of (2-3):(3-4):(1-1.5), with a total purity ≥95wt%, of which taurine purity ≥98wt%. The natural attractant is selected from salmon hydrolyzed peptides or taurine with a purity of ≥99wt%, wherein the molecular weight of the salmon hydrolyzed peptides is 500-1000 Da.

4. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, The double-layer embedding system includes: The mass ratio of sodium alginate to chitosan in the inner nanocarrier is 3:1-5:1, and the amount of cross-linking agent CaCl2 added is 0.5-1.5wt%; the mass ratio of L-carnosine encapsulated by liposomes is L-carnosine: dipalmitoylphosphatidylcholine = 1:1, the liposome particle size is 50-100nm, and the encapsulation efficiency is ≥95%; In the outer microcapsule wall material, the viscosity of gum arabic in a 10% aqueous solution at 25℃ is 220-320 mPa·s, the gel strength of gelatin is 230-260 Bloom and the ash content is ≤1.2wt%, the purity of glycerol is ≥99.5%, and the purity of β-cyclodextrin is ≥99% and the moisture content is ≤10wt%. The total encapsulation efficiency of the double-layer encapsulation system for the composite core material and L-carnosine is ≥92%.

5. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, The functional excipients include: Maltodextrin has a DE value of 12-18 and a moisture content of ≤5 wt%. The thermoresistant probiotic is Lactobacillus plantarum, with a survival rate of ≥90% and a viable count of ≥1×10⁻⁶ after being treated at 60℃ for 72 hours. 9 CFU / g; Sodium dihydrogen phosphate purity ≥99%, the pH of the system can be adjusted to 6.0-6.5; The amount of tert-butylhydroquinone added is 0.018-0.05 wt% of the total mass of the flavoring powder.

6. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, The double-layer encapsulation system achieves two-stage targeted release: the total release rate of the composite core material and L-carnosine within 2 hours under simulated cat gastric environment is ≤10%, and the total release rate of the composite core material and L-carnosine within 4 hours under simulated cat intestinal environment is ≥92%, and the difference between the intestinal release rate and the gastric release rate is ≥82%.

7. The cat microcapsule flavoring powder containing active plant ingredients according to claim 1, characterized in that, The microcapsule particle size of the seasoning powder is 10-25 μm; the moisture content of the seasoning powder is ≤5%, and the solubility at 25℃ is ≥92%; after storage at 25℃ and 65% relative humidity for 6 months, the retention rate of menthol is ≥88%, the retention rate of tea polyphenols is ≥85%, the retention rate of L-carnosine is ≥82%, and the survival rate of probiotics is ≥85%; after an accelerated test at 60℃ for 30 days, the retention rate of each active ingredient is ≥85%.

8. A method for preparing a cat microcapsule flavoring powder containing active plant ingredients as described in any one of claims 1-7, characterized in that, Includes the following steps: S1. Preparation of composite core material: Peppermint extract, green tea extract, chicory extract, mulberry leaf extract, and hawthorn extract are mixed in a mass ratio of (1.8-4.5):(1.2-3):(0.8-1.5):(0.5-1.2):(1-2) to obtain a composite core material; wherein the peppermint extract contains ≥18wt% menthol, the green tea extract contains ≥25wt% tea polyphenols, the chicory extract contains ≥60wt% inulin, the mulberry leaf extract contains ≥30wt% total flavonoids, and the hawthorn extract contains ≥15wt% triterpenic acid. S2. Construction of the inner nanocarrier: ① Prepare L-carnosine liposomes by mixing L-carnosine and dipalmitoylphosphatidylcholine at a mass ratio of 1:1 and dispersing them in PBS buffer at pH 7.4; ② Mix 2% sodium alginate solution and 1% chitosan solution at a mass ratio of 3-5:1, add 0.5-1.5wt% CaCl2 solution, and solidify by electrostatic adsorption reaction at 30℃ for 30±2min to obtain composite microspheres; ③ Add the composite core material and L-carnosine liposomes to the composite microspheres at a mass ratio of 1:(0.3-0.5), and stir at 35-40℃ and 120-150r / min for 30-40min to obtain the nanocarrier-active ingredient complex; S3. Preparation of outer microcapsule wall material: Weigh gum arabic, gelatin, and glycerin according to the proportion, add deionized water, stir at 65-72℃ until dissolved, cool to 38-42℃, add β-cyclodextrin, stir for 25-30 min to obtain wall material composite solution, the solid content of the wall material composite solution is 18-22 wt%; S4. Double-layer embedding and spray drying: The nanocarrier-active ingredient complex is added dropwise to the wall material composite solution at a volume ratio of 1:(2.5-3.5), stirred at 38-42℃ and 80-100 r / min for 1.2-1.8 h, the pH is adjusted to 4.2-4.6, and stirring is continued for 40-50 min to obtain a suspension; the suspension is spray dried with the following parameters: inlet air temperature 190-210℃, outlet air temperature 85-92℃, feed rate 18-22 mL / min, atomization pressure 0.25-0.35 MPa, and the dried powder is collected; S5. Mixing of excipients: Add functional excipients to the dry powder and stir at 120-150 r / min for 18-25 min to obtain cat microcapsule flavoring powder containing active plant ingredients.

9. The preparation method according to claim 8, characterized in that, In step S4, the inlet air temperature of the spray dryer is controlled at 195-205℃ and the outlet air temperature is controlled at 87-90℃; in step S5, the stirring speed is adjusted to 130-140 r / min and the stirring time is extended to 22-25 min.

10. The application of a cat microcapsule flavoring powder containing active plant ingredients as described in any one of claims 1-7, characterized in that, The seasoning powder is added at an amount of 1.5-3% of the total mass of the cat functional food to prepare the cat functional food. The cat functional food is selected from adult cat extruded dry food, senior cat low-temperature baked dry food, obese cat low-fat canned food, or postoperative cat nutritional freeze-dried meat strips. The cat functional food has a shelf life of 12 months under the conditions of 25°C and 65% relative humidity.

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