Process for preparing food composition for pet dog comprising as main component fermented sprout jinseng powder

Abstract

The present invention relates to a food composition for dogs comprising fermented sprout ginseng as an active ingredient, a method for manufacturing the same, and a method for manufacturing said fermented sprout ginseng extract. Since the food composition for dogs according to the present invention can provide not only palatability but also nutritional value and functional properties such as antioxidant properties, it can be appropriately consumed as a premium feed.

Description

Technology Field

[0001] The present invention relates to a food composition for companion dogs comprising a fermented sprout ginseng extract as an active ingredient, a method for manufacturing the same, and a method for manufacturing the fermented sprout ginseng extract. Background Technology

[0002] With the improvement of people's living standards and the increase in single-person households, the number of people raising pet dogs is rapidly increasing. Recently, these dogs are recognized as companion dogs, unlike in the past when they were fed leftover food, and now it has become common to feed them dog food.

[0003] Dry feed refers to a type of feed made by processing raw materials into pellet form to create a hard shape that is easy to store and eat; as the name suggests, it is feed with almost no moisture. Wet feed is a product that contains an appropriate amount of moisture, sold in cans or airtight containers after undergoing a simple cooking process that preserves the raw materials almost as they are. Dehydrated feed is a feed processed by completely removing the moisture from each raw material to make it almost dry; it has a moisture content of about 10%, has a long shelf life, is convenient to store, and transforms into a wet feed form as the raw materials absorb the water when water is added.

[0004] Many technologies regarding pet food are known, for example, Korean Patent No. 10-1346696 (December 24, 2013) discloses a method for manufacturing pellet-type dog food by processing salmon, pollack heads, sweet potatoes, spinach, carrots, and rosemary through a series of physical processes.

[0005] In addition, Korean Registration No. 10-2233957 (March 24, 2021) discloses a high-quality dog ​​food utilizing agricultural and livestock products and a method for manufacturing the same, which is characterized by comprising, for every 100 parts by weight of chicken breast, 22 parts by weight of egg, 8 parts by weight of corn powder, 8 parts by weight of soybean meal powder, 5 parts by weight of beet pulp powder, 9 parts by weight of wheat flour, 5 parts by weight of rice flour, 5 parts by weight of fruit jam, 1.2 parts by weight of salt, 11 to 50 parts by weight of a vegetable mixture in which 1.5 to 10 parts by weight each of sweet potato, carrot, and chives are mixed, and 3 parts by weight of TCP (tri-calcium phosphate).

[0006] According to recent import and export trade statistics from the Korea Customs Service regarding the status of domestic pet food imports and exports, the volume of exported domestic products is negligible, suggesting that domestic products currently play a role in the low-price pet food market. However, to survive and secure a competitive edge in the pet food market, which is expected to experience continuous growth, it is predicted that high-quality products—specifically those of edible quality—will generate high added value and gain prominence.

[0007] Therefore, there is an urgent need to develop super premium pet dog food that replaces imported feed, which accounts for a significant portion of the rapidly expanding pet market, by utilizing fresh, high-quality, and nutritious domestic agricultural and livestock products as primary ingredients. Prior art literature

[0008] Republic of Korea Registration No. 10-1346696 (December 24, 2013) Republic of Korea Registration No. 10-2233957 (March 24, 2021) The problem to be solved

[0009] As a result of diligent research to resolve the problems of the prior art and develop a high-quality feed for companion dogs, the inventors confirmed that a composition containing fermented sprout ginseng extract, as described below, can satisfy the requirements by providing functional properties such as antioxidants while also satisfying the nutritional needs and palatability of companion dogs, and thus completed the present invention. means of solving the problem

[0010] Therefore, the objective of the present invention is to provide a premium food composition for dogs that possesses not only palatability but also nutritional value and functional properties such as antioxidant properties.

[0011] The objective of the present invention as such is, in one aspect,

[0012] a) a step of purchasing raw materials and performing pretreatment including sorting;

[0013] b) A step of receiving, storing, and weighing the pre-treated raw materials;

[0014] c) A step of pre-treating animal-based food ingredients among the raw materials by steaming them at around 100℃ for 30 minutes;

[0015] d) Separately, a step of preparing a fermented sprout ginseng product;

[0016] e) A step of mixing individually prepared materials in a fixed ratio and then stirring thoroughly to mix them;

[0017] f) A step of compressing the mixture and then molding it into dog food of a specific shape;

[0018] g) A step of drying the molded feed at a temperature of around 75℃ for 1 hour;

[0019] h) a step of removing foreign substances from dried feed and sorting; and

[0020] This can be achieved by a method for manufacturing a food composition for companion dogs containing fermented sprout ginseng as an active ingredient, characterized by including the step of packaging selected feed into a certain volume. Effects of the invention

[0021] The food composition for dogs according to the present invention can provide not only palatability but also nutritional value and functional properties such as antioxidants, so it can be appropriately consumed as a premium feed. Brief explanation of the drawing

[0022] FIG. 1 is a manufacturing process diagram of a food composition for companion dogs containing fermented sprout ginseng as an active ingredient according to the present invention. FIG. 2 is (A): Lactobacillus Brevis Confirmation of OPK-3 GABA conversion activity TLC, and (B): Weissella Koreensis This is a TLC image confirming the ornithine conversion activity of OK1-6. Fig. 3 is (A): Lactobacillus Brevis OPK-3 (10 5 ) (B): Lactobacillus Brevis OPK-3 (10 6 ) (C): Weissella Koreensis OK1-6 (10 5 ) (D): Weissella Koreensis OK1-6 (10 6This is a culture photograph of ). Fig. 4 is a photograph showing the prepared sprout ginseng culture solution containing functional lactic acid bacteria. Fig. 5 is a TLC photograph of GABA activity of the fermented sprout ginseng material. Fig. 6 is a TLC photograph of Ornithine activity of the fermented sprout ginseng material. Fig. 7 is a TLC photograph of Ginsenoside change of the fermented sprout ginseng material. Fig. 8 shows the GABA HPLC analysis chromatogram of the fermented sprout ginseng material. Fig. 9 shows the Ornithine analysis chromatogram of the fermented sprout ginseng material. Figure 10 is a graph showing the GABA and Ornithine content of the fermented sprout ginseng material through HPLC analysis. Figure 11 shows the extraction and fractionation of the fermented sprout ginseng material. Figure 12 shows the results of measuring the DPPH free radical scavenging activity of the ethanol extract and fractions of the fermented sprout ginseng material. Figure 13 shows the results of measuring the ABTS radical scavenging activity of the ethanol extract and fractions of the fermented sprout ginseng material. Figure 14 is a graph showing the effect of the methylene chloride fraction of the fermented sprout ginseng material on antioxidant enzyme activity in Caenorhabditis elegans. Figure 15 is a graph showing the effect of the methylene chloride fraction of the fermented sprout ginseng material on ROS accumulation in Caenorhabditis elegans. Figure 16 is a photograph and graph showing the effect of the methylene chloride fraction of the fermented sprout ginseng material on SOD-3::GFP expression in transformed CF1553. Figure 17 shows the extracts and fractions of the fermented sprout ginseng material This shows the effect on extending the lifespan of Caenorhabditis elegans. Figure 18 is a graph showing the effect of the methylene chloride fraction of the fermented sprout ginseng material on the heat stress resistance of Caenorhabditis elegans. Figure 19 shows the effect of the methylene chloride fraction of the fermented sprout ginseng material on the accumulation of lipofuscin in Caenorhabditis elegans. Figure 20 shows the results of the nutritional component analysis of the premium fermented sprout ginseng material.Figure 21 shows the results of the analysis of moisture, ash, calcium, and phosphorus in fermented sprout ginseng. Figure 22 is a diagram showing the results of the sensory evaluation. Specific details for implementing the invention

[0023] The present invention, in one aspect,

[0024] a) a step of purchasing raw materials and performing pretreatment including sorting;

[0025] b) A step of receiving, storing, and weighing the pre-treated raw materials;

[0026] c) A step of pre-treating animal-based food ingredients among the raw materials by steaming them at around 100℃ for 30 minutes;

[0027] d) Separately, a step of preparing a fermented sprout ginseng product;

[0028] e) A step of mixing individually prepared materials in a fixed ratio and then stirring thoroughly to mix them;

[0029] f) A step of compressing the mixture and then molding it into dog food of a specific shape;

[0030] g) A step of drying the molded feed at a temperature of around 75℃ for 1 hour;

[0031] h) a step of removing foreign substances from dried feed and sorting; and

[0032] i) a step of packaging selected feed into a predetermined volume; the present invention provides a method for manufacturing a food composition for companion dogs containing fermented sprout ginseng as an active ingredient.

[0033] Hereinafter, a food composition for companion dogs containing fermented sprout ginseng as an active ingredient according to the present invention and a method for manufacturing the same will be described in more detail with reference to the attached drawings.

[0034] The terms and words used in this specification and claims are not limited to their ordinary or dictionary meanings, and must be interpreted in a meaning and concept consistent with the technical spirit of the invention, based on the principle that the inventor may appropriately define the concept of the terms to best describe his invention. Accordingly, since the embodiments described in this specification and the configurations illustrated in the drawings are merely the most preferred embodiments of the invention, it should be understood that various equivalents and modifications capable of replacing them may exist at the time of filing this application.

[0035] In the present invention, a food composition for dogs containing fermented sprout ginseng extract as an active ingredient is expressed in parts or weight percent, and may include an unfeasible portion in which the sum of individual ingredients exceeds 100 weight percent by a calculation formula. However, it is obvious that such excess portion or unfeasible portion is not included within the scope of the present invention. In particular, it is obvious that any amendment to clarify this in the patent claims, such as adding a proviso that the portion exceeding 100 weight percent is excluded from the scope of the present invention, merely clarifies unclear descriptions and does not affect the nature or scope of the present invention.

[0036] Figure 1 is a manufacturing process diagram of a food composition for dogs containing fermented sprout ginseng as an active ingredient according to the present invention.

[0037] A method for preparing a food composition for companion dogs comprising another fermented sprout ginseng extract as an active ingredient according to the present invention

[0038] a) a step of purchasing raw materials and performing pretreatment including sorting;

[0039] b) A step of receiving, storing, and weighing the pre-treated raw materials;

[0040] c) A step of pre-treating animal-based food ingredients among the raw materials by steaming them at around 100℃ for 30 minutes;

[0041] d) Separately, a step of manufacturing fermented sprout ginseng;

[0042] e) A step of mixing individually prepared materials in a fixed ratio and then stirring thoroughly to mix them;

[0043] f) A step of compressing the mixture and then molding it into dog food of a specific shape;

[0044] g) A step of drying the molded feed at a temperature of around 75℃ for 1 hour;

[0045] h) a step of removing foreign substances from dried feed and sorting; and

[0046] i) a step of packaging the selected feed into a certain volume; characterized by including

[0047] It may be desirable to utilize methods and equipment commonly used for feed production in the individual processes mentioned above, as needed.

[0048] To this end, the above-mentioned a) pretreatment step may include processes of inspecting and sorting the condition of materials containing grains and animal and plant components, removing foreign substances, washing and drying, and chopping as necessary.

[0049] It may be desirable to pre-treat animal-based food ingredients among the raw materials by steaming them at around 100°C for 30 minutes.

[0050] The step of d) manufacturing fermented sprout ginseng above

[0051] Step of washing, freeze-drying, and grinding young ginseng sprouts;

[0052] A step of preparing a culture solution by adding 30% ethanol to powdered ginseng sprouts, adding 1% sodium L(+)-glutamate monohydrate and L-arginine monohydrochloride, sterilizing by autoclaving, and then naturally cooling while sealed;

[0053] Step of inoculating microorganisms into a naturally cooled culture medium; and

[0054] It may be preferable to produce it by the step of fermenting a culture inoculated with microorganisms at 30°C for 72 hours.

[0055] The above microorganisms are Lactobacillus brevis OPK-3 and Weissella koreensis It may be preferable to have OK1-6. The above Lactobacillus brevis OPK-3 is a strain deposited as KFCC 11330, as disclosed in Korean Registration No. 10-0675955, etc., and Weissella koreensis OK1-6 is deposited as KFCC11469 and is disclosed in Korean registered patent 10-1130695, etc.

[0056] It may be preferable to use the above strain as a strain required for fermentation by inoculating it into MRS broth, culturing it at 30°C for 48 hours, subculturing it three times, and then subculturing 10 μL of the culture solution into skimmlk medium.

[0057] In a specific embodiment, washed young ginseng sprouts may be freeze-dried according to a conventional method and ground to a mesh size of 30 to 120 for use. Subsequently, based on 10 g of powdered fermented young ginseng sprouts, 100 mL of 30% ethanol is added, along with 1% sodium L(+)-glutamate monohydrate and L-arginine monohydrochloride; after autoclaving (121°C, 15 min), the mixture is sealed and naturally cooled to obtain a fermented liquid. Subsequently, Lactobacillus brevis OPK-3 and Weissella koreensis 0.5 g of OK1-6 lactic acid bacteria (OPK-3 is 9.4 log CFU / mL, OK1-6 is 9.3 log CFU / mL) is inoculated, and the fermentation liquid is prepared by fermenting at 30°C for 72 hours.

[0058] Next, in the e) mixing and stirring step, the individually prepared materials are mixed in a constant ratio and then stirred evenly.

[0059] The mixing ratio of the ingredients, based on parts by weight, preferably includes 17.65 parts salmon, 11.4 parts pesticide-free eco-friendly rice, 11.4 parts organic mixed grains, 11.4 parts sweet potato paste, 4.55 parts black bean powder, 4.55 parts pollack powder, 4.55 parts anchovy powder, 2.04 parts turmeric, 0.7 parts Korean angelica powder, 0.7 parts strawberry powder, 0.7 parts blueberry powder, 1.7 parts aronia powder, 1.7 parts barley sprout powder, 5.1 parts carrot powder, 0.7 parts spinach powder, 1.7 parts prickly pear powder, 5.1 parts floury rice, 5.1 parts puffed rice, 0.27 parts multivitamin, 4.55 parts fructooligosaccharide, 0.27 parts vegetable oil, 0.27 parts glasswort salt, 1.6 parts sprout ginseng powder, and 2.3 parts fermented sprout ginseng extract. there is.

[0060] The above-mentioned mixing ratio is obtained by considering the nutritional value and palatability of the ingredients. If the content falls outside the above range, there is a risk that the additive effect will be weak, the nutritional balance will be off, or harmony with each component will not be achieved, or the texture and taste will be inferior, or it may be uneconomical.

[0061] Accordingly, the technical feature of the present invention is that the materials and content defined above are materials that can harmonize useful components and unique aroma and nutritional components, and by finding the appropriate mixing ratio of these, the palatability and nutritional balance of the premium feed are found.

[0063] Examples

[0064] The present invention will be explained in more detail below with reference to examples. However, these examples are intended to illustrate the present invention and are not intended to limit the invention.

[0066] Example 1: Test to Confirm the Conversion Ability of Functional Microorganisms Producing GABA and Ornithine

[0067] Lactobacillus Brevis OPK-3 and Weissella Koreensis For the development of a functional fermented sprout ginseng material using OK1-6 Lactobacillus Brevis OPK-3 and Weissella KoreensisAfter culturing OK1-6 lactic acid bacteria in MRS broth containing 1% MSG and Arginine, the conversion ability to GABA and Ornithine was confirmed by Thin Layer Chromatography (TLC), and dilution plating was performed to confirm the growth pattern.

[0068] Figure 2 is (A): Lactobacillus Brevis Confirmation of OPK-3 GABA conversion activity TLC, and (B): Weissella Koreensis This is a TLC image confirming the ornithine conversion activity of OK1-6. Fig. 3 is (A): Lactobacillus Brevis OPK-3 (10 5 ) (B): Lactobacillus Brevis OPK-3 (10 6 ) (C): Weissella Koreensis OK1-6 (10 5 ) (D): Weissella Koreensis OK1-6 (10 6 This is a culture photo of ).

[0070] Example 2: Preparation of functional fermented material

[0071] The strain used for fermentation was inoculated into MRS broth and cultured at 30°C for 48 hours, then subcultured three times. 10 μL of the culture solution was subcultured into skimmlk medium and used as the strain required for fermentation.

[0072] To develop a functional fermented sprout ginseng material, washed sprout ginseng was freeze-dried and ground for use. 100 mL of 30% ethanol was added to 10 g of powdered sprout ginseng, along with 1% sodium L(+)-glutamate monohydrate and L-arginine monohydrochloride. After autoclaving (121℃, 15 min), the mixture was sealed and allowed to cool naturally (80 ml).

[0073] Next, Lactobacillus brevis OPK-3 and Weissella koreensis0.5 g of OK1-6 lactic acid bacteria (OPK-3 was 9.4 log CFU / mL, OK1-6 was 9.3 log CFU / mL) was inoculated, and the inoculum was fermented at 30°C for 72 hours to produce the product, which was then used for subsequent tests and production. Figure 4 is a photograph showing the prepared sprout ginseng culture solution containing functional lactic acid bacteria.

[0075] Example 3: Determination of Optimal Feed Formula Using Fermented Sprout Ginseng and Domestic Functional Materials

[0076] Market research on dog food and an investigation into the characteristics and efficacy of functional food ingredients suitable for dog food were conducted, and product development experiments were carried out using food ingredients that complement sprout ginseng and fermented sprout ginseng extract.

[0077] Three main concepts were established, and experiments were conducted by applying materials suitable for each concept. The three concepts were a healthy feed using salmon and fruits and vegetables, a high-protein feed using chicken and insect food, and a nourishing feed using duck and herbal ingredients. The recipe development, manufacturing, research, and experiments shown in Tables 1 to 3 and Figure 1 were conducted with factory system production (mass production) in mind.

[0078]

[0079]

[0081] Example 4: Preparation of Premium Dog Food

[0082] Dog food was prepared in the same manner as Example 3 above using the recipe shown in Table 3 below.

[0083] Material name Premium pet food manufacturing ratio (%) Control group pet food manufacturing ratio (%) note salmon 17.65 17.65 Pesticide-free eco-friendly rice 11.4 11.4 Organic mixed grains 11.4 11.4 Oats, barley, sorghum (equal weight ratio) Sweet potato paste 11.4 11.4 black bean powder 4.55 4.55 Pollack powder 4.55 4.55 anchovy powder 4.55 4.55 curcuma 2.04 2.04 Chrysanthemum powder 0.7 0.7 strawberry powder 0.7 0.7 blueberry powder 0.7 0.7 Aronia powder 1.7 1.7 barley sprout powder 1.7 1.7 Carrot powder 5.1 5.1 spinach powder 0.7 0.7 Prickly pear powder 1.7 1.7 Fluffy 5.1 5.1 Rice flour Puffed rice 5.1 5.1 Multivitamin 0.27 0.27 Vitamin C Fructooligosaccharides 4.55 4.55 vegetable oil 0.27 0.27 Canola oil glasswort salt 0.27 0.27 Sprout ginseng powder 1.6 3.9 Fermented sprout ginseng extract (Example 2) 2.3 - total 100 100

[0085] Test Example 1: Evaluation of GABA and Ornithine Production Ability of Functional Fermented Materials

[0086] 800 μL of extraction solvent (methanol : chlorform : water = 12 : 5 : 3) was mixed with 200 mg of fermented young ginseng sprout sample solution (Example 2), and after centrifugation (12,000 rpm, 15 min, 4℃), the supernatant (supernatant) was recovered.

[0087] 400 μL of chlorform and 200 μL of water were added to the lower layer (bottom liquid), mixed, and then centrifuged to recover the supernatant.

[0088] The first and second supernatants were combined and centrifuged (12,000 rpm, 15 min, 4℃). The supernatant was then freeze-dried, dissolved in ultrapure water, filtered, and used as a sample for TLC and HPLC analysis.

[0089] 1 μL of the sample was loaded onto a TLC silica gel and developed in a GABA developing solvent (butanol : acetic acid : water = 4 : 1 : 1) and an Ornithine developing solvent (butanol : acetic acid : dichloromethan : water = 5 : 3 : 3 : 3). The samples were stained with 2% ninhydrin, and the spot sizes of the standard and the sample spots were compared and analyzed. Figure 5 is a TLC image of the GABA activity of the fermented sprout ginseng material. Figure 6 is a TLC image of the Ornithine activity of the fermented sprout ginseng material.

[0091] Test Example 2: Evaluation of Saponin (Ginsenoside) Production Ability of Functional Fermented Materials

[0092] Methanol was added to the fermented sprout ginseng powder sample (Example 2) for sonication (10 min, repeated 3 times), followed by hot water extraction. After filtering the extract, ether was added to remove impurities (fat-soluble substances) (repeated 3 times). The extract layer was recovered, and an equal amount of water-saturated butanol was added to extract saponin (Ginsenoside) (repeated 3 times). The water-saturated butanol layers were combined, concentrated under reduced pressure, dissolved in methanol, filtered, and used as a sample for TLC analysis. 1 μL of the sample was loaded onto a TLC silica gel and developed in the developing solvent (chloroform : methanol : water = 6.5 : 3.5 : 1). The gel was stained with 10% sulfuric acid (EtOH), and the locations of the standard (Rb1, Rd, Fe, Rh2, CK) spots and the sample spots were compared and analyzed. Figure 7 is a TLC image showing the change in Ginsenoside in the fermented sprout ginseng material. CK stands for compound K among ginsenosides, and the arrow indicates the location of compound K.

[0094] Test Example 3: Measurement of GABA and Ornithine content in functional fermented sprout ginseng.

[0095] The GABA and ornithine content in fermented young ginseng sprouts was quantified by comparing amino acid chromatograms via HPLC analysis. For HPLC analysis, 70 μL of HPLC Reagent Kit (waters) was added to the extracted sample and reacted at 55°C for 10 minutes, after which HPLC analysis was performed. The HPLC instrument and setting conditions are as shown in Tables 4 and 5 below. Table 4 shows the HPLC instrument and analysis condition settings, and Table 5 shows the PLC analysis conditions.

[0096] Model Waters 2690(SIL-20AC) Column 3.9×150 mm AccQ.Tag TM(Nova-PakTM C18,Waters) column Mobile phase A = Waters eluent A : Water = 100:1000(v / v), pH=5.48B = 60% Acetonitrle Flow rate 1.0 ml / min Temperature 37℃ Detector Waters 747 scanning fluorescence detector Injection volume 10㎕

[0097] Time(min) Flow rate(ml / min) Mobile phase A % Mobile phase B % Init. 1.0 100 0 2.00 1.0 95 5 8.00 1.0 92.5 7.5 23.00 1.0 92 8 24.50 1.0 89 11 25.50 1.0 80.5 19.5 44.00 1.0 79.74 20.26 49.00 1.0 72 28 50.00 1.0 40 60 53.00 1.0 40 60 55.00 1.0 96 4 60.00 1.0 96 4

[0098] Figure 8 shows the GABA HPLC analysis chromatogram of the fermented sprout ginseng material. A: Overall chromatogram of the HPLC analysis of the fermented sprout ginseng material. B: Enlarged view of the GABA portion of the overall HPLC analysis chromatogram of the fermented sprout ginseng material.

[0099] Figure 9 shows the ornithine analysis chromatogram of the fermented sprout ginseng material. A: Overall HPLC analysis chromatogram of the fermented sprout ginseng material, B: Enlarged view of the ornithine portion of the HPLC analysis chromatogram of the fermented sprout ginseng material.

[0100] Figure 10 is a graph showing the GABA and Ornithine content of fermented sprout ginseng material through HPLC analysis.

[0102] Test Example 4: Measurement of saponin content in fermented young ginseng sprouts

[0103] 1 g of fermented sprout ginseng sample was extracted with 30 mL of methanol as a solvent for 1 hour, filtered, and extracted three times. The extracts were combined and concentrated, and the concentrate was dissolved in 50 mL of methanol as a solvent and used as a sample. 100 μL of the prepared sample was placed in a test tube, and 300 μL of 8% vaniline (EtOH) solution and 4 mL of 72% sulfuric acid solution were added in an ice bath. The test tube containing the solution was heated in a 60°C water bath for 10 minutes to induce color development, and the absorbance was measured at 545 nm. A calibration curve was constructed using Ginsenoide Rb1 as a standard solution, and calculations were performed according to the following formula.

[0104] Y (%) = Y' × {(T / V) / W} × 100 [wherein, Y': amount obtained by substituting the spectrophotometer of the test solution into the calibration curve (mg), T: volume of saponin extract dissolved to the correct volume (mL), V: volume taken during spectrophotometric measurement (μ L), W: sample amount (mg)]

[0105] The results are shown in Table 6 below. Table 6 shows the results of measuring the total saponin (Ginsenoside) content of the fermented sprout ginseng material.

[0106] Saponin level (g / 100g) FSG n -Hex MC EA n -BuOH 29.1 5.6 9.8 8.9 7.4

[0108] Test Example 5: Measurement of polyphenol content in fermented young ginseng sprouts

[0109] The polyphenol content in fermented young ginseng sprout samples was measured using an application of the Folin-Ciocalteau colorimetric method. 20 mg of the sample was diluted with 10 mL of ethanol and used as the sample. 100 μL of Folin-Ciocalteau solution was added to 100 μL of the sample and homogenized for 2 minutes. After 2 minutes, 800 μL of 0.1 M sodium carbocate was added, and the mixture was reacted in a 40°C water bath for 20 minutes to measure the absorbance at 700 nm. The polyphenol content was calculated as total phenol content (mg tannic acid equivalent (TAE) / g) using tannic acid as a standard and a linear equation of concentration (y = 0.0022x + 0.0093 R²=0.9981).

[0110] Extract and fraction Total polyphenol(μg TAE / mL) 1) Etanol extract 43.04 n -hexane fraction 114.55 methylene chloride fraction 123.18 ethyl acetate fraction 44.32 n -butanol fraction 22.64

[0112] Test Example 6: Analysis of Antioxidant / Anti-aging Efficacy of Premium Fermented Sprout Ginseng Material

[0113] 6-1) Development of Manufacturing Process for Fermented Sprout Ginseng Powder and Preparation of Extracts (Fractions)

[0114] In order to investigate the antioxidant and anti-aging efficacy of fermented sprout ginseng using functional lactic acid bacteria, fermented sprout ginseng powder was solvent-extracted and concentrated under reduced pressure to obtain a concentrate, which was then suspended in water, and an equal amount n -hexane (0.21 g), methylene chlordie (0.25 g), ethyl acetate (0.24 g), n Each fraction was obtained by sequentially solvent fractionating with -butanol (0.4 g). Figure 11 shows the extraction and fractionation of the fermented sprout ginseng material.

[0116] 6-2) DPPH free radical scavenging activity of fermented sprout ginseng material

[0117] To measure the DPPH free radical scavenging activity of fermented sprout ginseng, 50 μL of samples prepared at different concentrations (250, 500 μg / mL) using ethanol as a solvent were mixed with 200 μL of 0.2 mM 1,1-diphenyl1-2-picrylhydrazyl (DPPH) solution and reacted for 30 minutes in the dark at room temperature. The absorbance was then measured at 517 nm using a microplate reader. L-ascorbic acid was used as a control drug, and the experiment was repeated three times.

[0118] The results are shown in Fig. 12 and Table 8. Fig. 12 shows the results of measuring the DPPH free radical scavenging activity of the ethanol extracts and fractions of the fermented sprout ginseng material, and Table 5 shows the DPPH free radical IC50 of the ethanol extracts and fractions of the fermented sprout ginseng material. 50 It is a value.

[0119] EtOH n -Hex MC EtOAc n -BuOH IC 50 (mg / mL) 14.11 3.29 0.40 3.19 19.81

[0120] The results of measuring the DPPH free radical scavenging activity of fermented sprout ginseng materials showed that the radical scavenging activity of fermented sprout ginseng was relatively superior compared to unfermented sprout ginseng, and it was confirmed that the scavenging activity increased in a concentration-dependent manner. The radical scavenging ability for each sample is as follows, and the ethanol extract (IC2) 50 value, 14.11 mg / mL), n -hexane fraction (IC 50 value, 3.29 mg / mL), methylene chloride fraction (IC 50 value, 0.40 mg / mL), ethyl acetate fraction (IC 50 value, 3.19 mg / mL), n -butanol fraction (IC 50The radical scavenging activity of the methylene chloride fraction was measured to be the best with a value of 19.81 mg / mL.

[0121] 6-3) ABTS radical scavenging activity of fermented sprout ginseng material

[0122] To measure the ABTS radical scavenging activity of fermented sprout ginseng materials, equal amounts of ABTS (7.4 mM) solution and K2S2O8 (2.6 mM) solution were mixed and reacted in a dark place at room temperature for 24 hours to induce radicals. The solution was then diluted with phosphate buffer saline (pH 7.4) to achieve an absorbance value of 0.7 ± 0.03. 10 μL of each sample at different concentrations was mixed with 190 μL of ABTS solution and reacted in a dark place at room temperature for 10 minutes, after which the absorbance was measured at 732 nm. The results of the ABTS radical scavenging activity measurement of the fermented sprout ginseng materials are shown in Figure 13 and Table 6.

[0123] Figure 13 shows the results of measuring the ABTS radical scavenging activity of ethanol extracts and fractions of fermented sprout ginseng material, and Table 9 shows the ABTS radical IC50 of ethanol extracts and fractions of fermented sprout ginseng material. 50 It is a value.

[0124] EtOH n -Hex MC EtOAc n -BuOH IC 50 (mg / mL) 1.41 37.17 0.15 0.16 1.69

[0125] The radical scavenging activity of fermented ginseng sprouts increased slightly compared to unfermented ginseng sprouts, and regarding the radical scavenging ability of each sample, the scavenging activity of the methylene chloride fraction was measured to be the best as follows. ethanol extract (IC 50 value, 1.41 mg / mL), n -hexane fraction (IC 50 value, 37.17 mg / mL), methylene chloride fraction (IC 50 value, 0.15 mg / mL), ethyl acetate fraction (IC50 value, 0.15 mg / mL), n -butanol fraction (IC 50 value, 1.69 mg / mL)

[0127] 6-4) Antioxidant enzyme (SOD, catalase) activity of the methylene chloride fraction of fermented sprout ginseng material in Caenorhabditis elegans

[0128] For the measurement of antioxidant enzyme activity of the methylene chloride fraction of fermented sprout ginseng material in Caenorhabditis elegans, nematodes at the same growth stage were cultured on plates prepared at different concentrations (250 and 500 μg / mL). On the second day, the nematodes were washed three times with M9 buffer, ground, and used for enzyme activity measurement. The composition of the homogenization buffer used for ground pulverization was 10 mM Tris-HCl, 150 mM NaCl, 0.1 mM EDTA, and pH 7.5. For superoxide dismutase (SOD) activity, a reaction mixture (1.6 mM xanthine and 0.49 mL of 0.48 mM NBT) was prepared using 10 mM phosphate buffer (pH 8.0) as the solvent, and then 10 μL of each concentration sample was added and per-incubated at 37°C for 5 minutes. Subsequently, xanthine oxidase was added and incubated at 37°C for 20 minutes, after which the reaction was stopped with 69 mM SDS and the absorbance was measured at 570 nm. The catalase activity of the methylene chloride fraction of fermented young ginseng sprouts was measured by reacting 50 μL of samples at different concentrations in 25 mM H2O2 for 3 minutes and measuring the absorbance at 240 nm, and the results are shown in Fig. 14. Fig. 14 is a graph showing the effect of the methylene chloride fraction of fermented young ginseng sprouts on the antioxidant enzyme activity in Caenorhabditis elegans.

[0129] As a result of measuring the antioxidant enzyme activity of the fermented sprout ginseng methylene chloride fraction in Caenorhabditis elegans, the SOD activity in the group administered 250 μg / mL of the fermented sprout ginseng methylene chloride fraction increased SOD activity by 24.44% compared to the control group, and the group administered 500 μg / mL increased SOD activity by 41.70%, as shown in the figure below. As for catalase activity, the group administered 250 μg / mL of the fermented sprout ginseng methylene chloride fraction increased activity by 18.65% compared to the control group, and the group administered 500 μg / mL increased activity by 44.38%, as shown in the figure below.

[0130] 6-5) Efficacy of the methylene chloride fraction of fermented sprout ginseng material in reducing ROS in Caenorhabditis elegans

[0131] Reactive oxygen species (ROS) within nematode cells of the methylene chloride fraction of fermented young ginseng were measured using 2'7'-dichlordihydrofluorescein diacetate (H2DCF-DA). Nematodes at the same growth stage were cultured in plates prepared with different concentrations of the methylene chloride fraction of fermented young ginseng. On the fourth day after reaching adulthood, they were placed in M9 buffer containing 100 μM juglone and incubated for 2 hours; subsequently, 5 nematodes were transferred to 50 μL of M9 buffer in a 96-well plate. Finally, 50 μL of 50 μM H2DCF-DA was added, and fluorescence intensities were measured at excitation 485 mM and emission 535 nm, respectively. The results are shown in Fig. 15. Fig. 15 is a graph showing the effect of the methylene chloride fraction of fermented young ginseng on ROS accumulation in Caenorhabditis elegans. As a result of measurement, as shown in the figure below, the decrease in fluorescence up to 120 minutes due to reactive oxygen species was reduced by approximately 14.68% in the group administered 250 μg / mL of fermented sprout ginseng methylene chloride fraction and by approximately 18.5% in the group administered 500 μg / mL when compared to the control group.

[0133] 6-6) Activity of methylene chloride fraction of fermented sprout ginseng material to increase SOD-expression in Caenorhabditis elegans

[0134] To evaluate the activity of the methylene chloride fraction of fermented sprout ginseng material in increasing SOD-3::GFP expression in Caenorhabditis elegans, CF1553 nematodes containing transformed SOD-3::GFP were cultured in media administered at various concentrations. The nematodes were used on the third day after reaching adulthood, and the nematodes were anesthetized with sodium azide (4%). GFP expression was observed using a fluorescence stereomicroscope (Olympus, Tokyo, Japan). To quantify and analyze the expression intensity, photographs were taken using the microscope and analyzed using Image J software. Figure 16 shows photographs and graphs illustrating the effect of the methylene chloride fraction of fermented sprout ginseng material on SOD-3::GFP expression in transformed CF1553. As a result of experiments using the transgenic nematode CF1533 containing SOD-3, the group treated with 250 μg / mL of the methylene chloride fraction of the fermented sprout ginseng material showed a 7.1% increase in GFP expression compared to the control group, and the group treated with 500 μg / mL showed a 14.29% increase in SOD-3::GFP expression.

[0136] 6-7) Evaluation of Shelf Life Extension Efficacy of Methylene Chloride Fraction from Fermented Young Ginseng Sprouts

[0137] To evaluate the lifespan-extending efficacy of the methylene chloride fraction of fermented sprout ginseng, eggs were isolated from NGM plates and transferred to plates supplemented with the fermented sprout ginseng fraction (250 μg / mL, 500 μg / mL) at the same nematode growth stage, where they were cultured (20℃), and survival was checked daily. The confirmation of survival was C. elegans The results were confirmed by the reaction when stimulated with the tip of a platinum wire, and all experiments were repeated three times. Figure 17 shows the effects of fermented sprout ginseng extracts and fractions on the lifespan extension of Caenorhabditis elegans, and Table 10 shows the effects of fermented sprout ginseng extracts and fractions on the lifespan extension of Caenorhabditis elegans.

[0138] Fraction Mean lifespan(day) Maximun lifespan(day) Change in meanlifespan(%) Log-rank test Control 9.4 ± 0.2 16 - - Ethanol 9.9 ± 0.2 16 4.7 - n -Hexnae 9.4 ± 0.2 17 - - Methylene Chloride 10.8 ± 0.2 20 15.2 *** p<0.001 Ethyl acetate 10 ± 0.2 18 6.6 ** p<0.01 n -Butanol 9.6 ± 0.2 18 2.3 -

[0139] As a result of confirming the effects of fermented sprout ginseng extract and fractions on the lifespan of nematodes, n Excluding the -hexnae fraction, the ethanol extract was 4.67%, the methylene chloride fraction was 15.24%, and the ethyl acetate fraction was 6.61%. n The -butanol fraction showed a 2.30% increased lifespan extension effect, indicating that the methylene chloride fraction significantly increased the lifespan of nematodes. The average lifespan of control nematodes was 9.4±0.2 hours, while the average lifespan of nematodes treated with the fermented young ginseng sprout methylene chloride fraction was 10.8±0.2 hours, demonstrating a 15.24% increase in lifespan extension efficacy. (15.2% *** p<0.001 )

[0141] 6-8) Heat shock resistance efficacy of the methylene chloride fraction of fermented sprout ginseng material

[0142] To evaluate the heat shock resistance of the methylene chloride fraction of fermented ginseng sprout material, nematodes were cultured on each plate supplemented with the fermented ginseng sprout methylene chloride fraction (250 μg / mL, 500 μg / mL). To confirm heat shock resistance, the nematodes were cultured at 36°C, a high-temperature condition, on the 4th day of adulthood, and the survival rate was measured every hour.

[0143] Figure 18 is a graph showing the effect of the methylene chloride fraction of fermented sprout ginseng material on the heat stress resistance of Caenorhabditis elegans. Table 11 shows the effect of the methylene chloride fraction of fermented sprout ginseng material on the heat stress resistance of Caenorhabditis elegans.

[0144] Stress condition Fraction Mean lifespan(h) Maximunlifespan(h) Chanfee in mean lifespan(%) Log-rank test 36°C thermal tolerance Control 7.8 ± 0.6 13 - - 250μg / mL 9.5 ± 0.6 15 22 * p<0.05 500μg / mL 9.9 ± 0.6 16 26.7 ** p<0.01

[0145] When the survival rates were compared by applying high-temperature stress of 36°C to nematodes that survive stably at 20°C, the control group died after 3 hours and all died by 13 hours, whereas the groups administered 250 and 500 μg / mL of fermented sprout ginseng methylene chloride fraction died starting from the 4th hour and survived until 15 and 16 hours, respectively. The average survival time of the control group nematodes was measured to be 7.8±0.6 hours, the average survival time of the 250 μg / mL administration group was 9.5±0.6 hours, showing a survival rate that increased by 22%, and the average survival time of the 500 μg / mL administration group was 9.9±0.6 hours, showing a survival rate that increased by 26.7%.

[0147] 6-9) Efficacy of the methylene chloride fraction of fermented sprout ginseng on the accumulation of aging pigment (lipofuscin) in nematodes

[0148] To evaluate the efficacy of the methylene chloride fraction of fermented sprout ginseng on the accumulation of senescence pigments in nematodes, N2 nematodes at the same growth stage were cultured in plates of different concentrations. On the 8th day of adulthood, nematodes were anesthetized with sodium axide (4%), and the accumulation of the autofluorescent senescence pigment lipofuscin in the nematodes was observed using a fluorescence microscope (Olympus, Japan). The expression intensity of GFP was measured using Image J software (USA). Figure 19 shows the effect of the methylene chloride fraction of fermented sprout ginseng on the accumulation of the senescence pigment (lipofuscin) in Caenorhabditis elegans.

[0149] The effect of the methylene chloride fraction of fermented sprout ginseng on the accumulation of aging pigments in nematodes was investigated by measuring the fluorescence expression using N2 nematodes on the 8th day of adulthood. The results showed that the group administered 250 μg / mL of the fermented sprout ginseng methylene chloride fraction reduced the accumulation of lipofuscin by 6.72%, while the group administered 500 μg / mL reduced the accumulation of lipofuscin by 7.58%.

[0150] Test Example 7: Nutritional Analysis of Premium Fermented Sprout Ginseng Material

[0151] The nutrient analysis of the fermented sprout ginseng material was commissioned to an external testing agency, and the results are shown in Fig. 20. Fig. 20 shows the results of the nutritional analysis of the premium fermented sprout ginseng material. The test results showed that the nutrients were contained in the order of sodium, carbohydrates, and protein.

[0152] In addition, the results of the analysis of moisture, ash, calcium, and phosphorus in fermented sprout ginseng are shown in Figure 21. Figure 21 shows the results of the analysis of moisture, ash, calcium, and phosphorus in fermented sprout ginseng. As a result, it contained phosphorus, calcium, moisture, and ash in that order.

[0153] Test Example 8: Physicochemical Analysis of Premium Dog Food

[0154] Physicochemical analysis (sugar content, salinity, pH, and color) of feed prepared using fermented sprout ginseng and domestic functional materials was measured. Sugar content was measured by taking 1 mL of the sample, attaching it to a refractometer sensor, and calculating the average value through repeated measurements. Salinity was measured by taking 1 mL of the sample, attaching it to a salinity sensor, and calculating the average value through repeated measurements. Color was measured using a colorimeter, and the average value was calculated through repeated measurements. pH was measured using a pH meter, and the average value was calculated through repeated measurements. The results are shown in Tables 12 and 13.

[0155] Sweetness salinity pH 1st measurement 2.3 1.9 5.94 2nd measurement 2.3 1.9 5.96 3rd measurement 2.3 1.9 5.95 average value 2.30 1.90 5.95

[0156] L A B 1st measurement 1.41 4.6 2.43 2nd measurement 1.42 4.67 2.38 3rd measurement 1.4 4.67 2.41 average value 1.41 4.67 2.41

[0158] Test Example 9: Palatability Test of Feed Using Fermented Sprout Ginseng and Domestic Functional Materials

[0159] 9-1) Dog Food Palate Survey and Questionnaire Analysis

[0160] A preference survey was conducted using an observational method with 10 dog owners for the development of dog food. The survey was conducted over five days to analyze specific reactions to the developed product. A total of 10 questionnaires were distributed, and all respondents answered honestly, with the responses used for analysis.

[0161] To conduct this study, a questionnaire was developed by referring to existing literature and prior research, and the questionnaire was composed of three main parts.

[0162] First, to identify the basic characteristics of the dogs in the survey sample, the questionnaire consisted of general questions regarding the dog's breed, gender, age, duration of living with the dog, and breakfast feeding time.

[0163] Second, to investigate the amount of food consumed by the dogs, a questionnaire was completed regarding the amount of food given before eating and the amount remaining after eating for a total of 5 days from day 1 to day 5.

[0164] Third, for the sensory evaluation after feeding the prototype feed for 5 days, questions were prepared using a 7-point scale ranging from 'very good' to 'very bad' for 5 items including color, flavor, taste, chewiness, and overall acceptability.

[0165] The analysis of the basic data collected in this study was performed using the statistical analysis program (IBM SPSS Statistics 19), and statistical characteristics were analyzed through frequency analysis, cross-tabulation, and radial chart analysis. In addition, in this analysis, the significance level p The significance between items was verified at < .05.

[0166] The gender of the dogs was 6 males (60%) and 4 females (40%), and the age of the dogs was 4 dogs aged 3 years or younger (40.0%), 3 dogs aged 4-6 years (30.0%), 3 dogs aged 7-9 years (30.0%), and 0 dogs aged 10 years or older (0%).

[0167] Looking at the duration of time spent with the dogs, 5 dogs (50.0%) have lived for 3 years or less, 2 dogs (20.0%) for 4-6 years, 3 dogs (30.0%) for 7-9 years, and 0 dogs (0%) for 10 years or more. Regarding the time of morning feeding, 2 dogs (20%) were fed before 7:00, 0 dogs (0%) between 7:00 and 8:00, 3 dogs (30%) between 8:00 and 9:00, and 5 dogs (50%) after 9:00. Table 14 shows the results of the cross-analysis regarding the gender of the dogs and the remaining items.

[0168] item gender every χ² p man female Dog's age 3 years old and under 3(75.0) 1) 1(57.1) 4(100) χ² =3.635( p = .304) 4 - 6 years old 2(66.7) 1(33.3) 3(100) Ages 7 - 9 1(33.3) 2(66.7) 3(100) Duration of living with a pet dog 3 years or less 3(60.0) 2(40.0) 5(100) χ² =6.200( p = .102) 4-6 years 2(100.0) 0(00.0) 2(100) 7-9 years 1(33.3) 2(66.7) 3(100) Dog's morning feeding time Before 7 o'clock 2(100.0) 0(00.0) 2(100) χ² =3.359( p = .339) 7 PM - 8 PM 1(33.3) 2(66.7) 3(100) 8 o'clock - 9 o'clock 3(66.7) 2(33.3) 5(100) every 6(60.0) 4(40.0) 10(100) 　 1) (%), N=10

[0170] 9-2) Analysis of Variance on Feed Intake

[0171] The average food intake of the dogs under investigation over 5 days was 46.0g on day 1, 44.0g on day 2, 46.5g on day 3, 44.5g on day 4, and 47.0g on day 5, and the F-analysis results showed that there was almost no difference in intake between the days .

[0172] intake cycle average Standard deviation F( p ) Day 1 46.0 1) 9.661 F=0.2240( p = .9236) Day 2 44.0 9.944 Day 3 46.5 7.472 Day 4 44.5 8.960 Day 5 47.0 6.749 1) (g), N=10

[0173] Analysis of feed intake results showed that all surveyed dogs consumed the experimental food well, consuming at least an average of 44g or more out of the daily allocated 50g of food over all five days, regardless of the round. Additionally, the difference in average intake per round was found not to be statistically significant.

[0174] 9-3) Results of Sensory Evaluation for Preference Survey

[0175] To investigate the palatability of the feed, the results of the analysis of color, flavor, taste, chewiness, and overall acceptance after feeding the feed for 5 days are shown in Fig. 22 and Table 16. Fig. 22 is a diagram showing the results of the sensory evaluation.

[0176] item average Standard deviation F( p ) Color 5.2 1.40 F=0.7313( p =0.1597) Flavor 5.3 1.34 Taste 6.4 1.27 Chewiness 6.0 0.94 Overall Acceptability 6.0 1.16

[0177] All five items showed excellent sensory evaluation results with an average of 5 or higher. Although the average for color was the lowest at 5.2, it still scored above 5, while the evaluation for taste showed the highest result at 6.4, indicating that the taste of the developed feed was preferred the most. Consequently, all dogs responded best to the taste, and responses to the remaining items were also generally excellent, so no statistically significant differences were found among the five items subject to sensory evaluation.

[0178] The results of the sensory evaluation, analyzed using a radial chart, show that most dogs, with the exception of the Welsh Corgi, respond well to all categories, with the response to taste being particularly excellent.

[0179] As such, the present invention has been described by way of example through the drawings and specification, but it is obvious to those skilled in the art that various modifications and equivalent implementations are possible.

Claims

Claim 1 a) a step of purchasing raw materials and performing pretreatment including sorting; b) a step of receiving, storing, and weighing the pretreated raw materials; c) a step of pretreating animal food ingredients among the raw materials by steaming them at around 100°C for 30 minutes; d) separately, a step of manufacturing fermented sprout ginseng; e) a step of mixing individually prepared materials in a certain ratio and mixing them by stirring evenly; f) a step of compressing the mixture and molding it into dog food of a certain shape; g) a step of drying the molded food at a temperature of around 75°C for 1 hour; h) a step of removing foreign substances from the dried food and sorting; and i) a step of packaging the sorted food in a certain volume; wherein the step d) manufacturing fermented sprout ginseng comprises a step of washing the sprout ginseng, freeze-drying it, and grinding it; The method is prepared by the steps of: adding 30% ethanol to powdered ginseng sprouts, adding 1% sodium L(+)-glutamate monohydrate and L-arginine monohydrochloride, sterilizing by autoclaving, and then naturally cooling while sealed to prepare a culture medium; inoculating the naturally cooled culture medium with microorganisms; and fermenting the culture medium inoculated with microorganisms at 30°C for 72 hours; wherein the microorganisms Lactobacillus brevis OPK-3 and Weissella koreensis A method for preparing a food composition for dogs containing a fermented sprout ginseng extract characterized by OK1-6 as an active ingredient. Claim 2 delete Claim 3 delete Claim 4 In claim 1, the composition comprises, based on weight parts, 17.65 parts salmon, 11.4 parts pesticide-free eco-friendly rice, 11.4 parts organic mixed grains, 11.4 parts sweet potato paste, 4.55 parts black bean powder, 4.55 parts pollack powder, 4.55 parts anchovy powder, 2.04 parts turmeric, 0.7 parts Korean angelica powder, 0.7 parts strawberry powder, 0.7 parts blueberry powder, 1.7 parts aronia powder, 1.7 parts barley sprout powder, 5.1 parts carrot powder, 0.7 parts spinach powder, 1.7 parts prickly pear powder, 5.1 parts floury rice, 5.1 parts puffed rice, 0.27 parts multivitamin, 4.55 parts fructooligosaccharide, 0.27 parts vegetable oil, 0.27 parts glasswort salt, 1.6 parts sprout ginseng powder, and 2.3 parts fermented sprout ginseng extract. A method for preparing a food composition for dogs containing a fermented sprout ginseng extract as an active ingredient, characterized by including.

Images