Cooked pet food composition comprising seaweed and manufacturing method therefor

Abstract

The present invention relates to a cooked pet food composition comprising salt-removed seaweed, and a manufacturing method therefor. Specifically, the present invention relates to: a cooked pet food composition comprising seaweed removed of salt by repeating, three times, a step of immersing dried seaweed in 30 times the weight of water for 18 to 22 minutes and removing the water or repeating, twice, a step of immersing dried seaweed in 40 times the weight of water for 18 to 22 minutes and removing the water; and a manufacturing method therefor. The cooked pet food composition according to the present invention comprises seaweed removed of salt, and thus can supply useful ingredients of seaweed to a pet without causing salt-induced harm to the health of the pet, and can be consumed by the pet without repulsion.

Description

Compound of seaweed-containing pet food and method for producing the same

[0001] The present invention is a result of a study on the development of pet food with enhanced functionality and physiological activity, which is a research project of Kyungsung University (Project No. 202300880001), and relates to a pet food composition containing seaweed and a method for manufacturing the same. Specifically, the present invention relates to a pet food containing desalinated seaweed or laver as an active ingredient, and a method for manufacturing a pet food, the method comprising the step of removing the salt from at least one type of seaweed selected from the group consisting of seaweed and laver, drying and powdering the same, and adding the same to a pet food.

[0002] Due to factors such as the increasing number of nuclear families, an aging population, and the rise of single-person households, the number of households owning pets, which provide psychological stability, is steadily increasing. In particular, the number of companion dogs increased by 29.7% over nine years, from 4.61 million in 2010 to 5.98 million in 2019.

[0003] Due to the increase in the number of companion animals, the production value of pet food in 2019 increased by 24.6% year-on-year to KRW 365.6 billion, and the shipment value in the same year increased by 24.7% year-on-year to KRW 365 billion, showing that the production performance of pet food is maintaining a steady increase.

[0004] With the ongoing trend of pet humanization, the premium pet food market, which utilizes high-quality ingredients, is growing. In particular, premium dry pet food accounts for a significant portion of the US pet food market, at 31.1% of the total. Furthermore, product differentiation and quality improvement are driving changes in pet food products to meet consumer needs for nutrition, functionality, and safety. Demand is particularly strong for functional products that support skin and coat health, intestinal and stomach health, joint and cartilage strengthening, natural, plant-based, and organic ingredients, and weight control. Furthermore, interest is growing in limited ingredient diet (LID) products aimed at allergy prevention and products using eco-friendly packaging.

[0005] Accordingly, pet food is not just simple feed, but, like food, includes the concept of differentiated consumption based on product quality, function, and purpose, and there is a need to develop various products to meet consumption patterns.

[0006] For example, obesity rates in companion animals have been increasing recently, and obesity can cause various diseases in companion animals, such as cardiovascular disease, diabetes, and degenerative arthritis. Furthermore, according to the Ministry of Agriculture, Food and Rural Affairs of Korea, senior dogs aged 7 years or older accounted for a high proportion of all registered companion dogs between 2008 and December 2017, at 45.56%. Consumer interest in cognitive and immune-boosting feed for senior dogs is high. To address these issues, customized feeds tailored to the health of companion animals or functional feeds for special purposes are being developed and released. Consumer interest in the functionality, nutritional value, and safety of feed is also on the rise.

[0007] Seaweeds such as kelp, wakame, and laver are low-calorie foods rich in dietary fibers such as alginic acid, which can help prevent cardiovascular disease, lower cholesterol, and obesity. In addition, seaweed is rich in minerals (calcium, iron, iodine, magnesium, phosphorus, potassium, zinc, copper, manganese, selenium, and fluorine), vitamins (A, B1, B2, B9, B12, C, D, E, and K), and polyphenols, which help with growth and development, and have antioxidant effects, including preventing osteomalacia and osteoporosis. In addition, fucoidan, a polysaccharide contained in brown algae such as wakame and kelp, has almost no toxicity or side effects when consumed, and has various biological functions such as inducing apoptosis in cancer cells to prevent cancer.

[0008] Therefore, there have been various attempts to provide useful components of seaweed as animal feed. For example, Korean Patent No. 10-0401486 (announced on October 10, 2003) discloses a complete mixed feed for producing Korean beef containing seaweed by-products, which is composed of 68.58 wt% of ground corn, 9.60 wt% of wheat bran, 6.67 wt% of soybean meal, 3.33 wt% of soybean hull, 1.67 wt% of limestone, 3.33 wt% of fermented sawdust, 3.33 wt% of sesame cake, 0.49 wt% of vitamin and mineral complex, and 3.0 wt% of seaweed by-products, and Korean Patent No. 10-1556887 discloses a complete mixed feed for producing Korean beef containing seaweed by-products, which is composed of 8-10 wt% of seaweed ears and kelp stems, 10-12 wt% of corn, 5.5-6.5 wt% of cottonseed, 7-8 wt% of protein hull, and 8.5-9.5 wt% of corn DDGS. Disclosed is a cattle feed composition comprising: 13 to 14 wt% of brewer's grain, 2.5 to 3.5 wt% of molasses, 8.5 to 9.5 wt% of alfalfa, 4.5 to 5.5 wt% of oat hay, 1.5 to 2.5 wt% of timothy hay, 3.5 to 4.5 wt% of perennial ryegrass, 1.0 to 1.25 wt% of fermented feed, 0.5 to 0.9 wt% of limestone, 0.15 to 0.25 wt% of salt, 0.1 to 0.2 wt% of baking soda, 0.1 to 0.25 wt% of protected fat, and 13 to 17 wt% of fermented water, and Korean Patent No. 10-2404021 discloses a cattle feed composition comprising: 30 to 45 wt% of seaweed, 30 to 35 wt% of tannin, 13 to 18 wt% of MSG, 7 to 10 wt% of complex probiotic, and sugar. A feed additive using seaweed containing 3 to 4 wt% of cellulose and 2 to 3 wt% of salt is disclosed.

[0009] However, when feeding the seaweed to animals as feed or feed additives, the salt content cannot be increased, and without devising a special method for removing salt, the salt has been removed only to the extent of washing the seaweed to remove foreign substances. Therefore, when applying the previously known compositions and methods to pets, the high salt content of the seaweed can have a negative effect on the pet's kidneys, etc., and thus, there is a problem in using it as pet food, especially as animal food.

[0010] Accordingly, the inventors of the present invention first developed a method for removing salt from seaweed in order to develop a pet food containing seaweed, and confirmed that seaweed from which salt was removed by the method has a very low salinity and is therefore suitable as pet food, thereby completing the present invention.

[0011] One object of the present invention is to provide a method for removing salt from seaweed, which comprises a step of immersing dried seaweed in water to remove salt.

[0012] Another object of the present invention is to provide a pet food composition comprising seaweed from which the salt has been removed.

[0013] Another object of the present invention is to provide a method for producing a pet food composition comprising seaweed from which the salt has been removed.

[0014] In one aspect, the present invention provides a method for removing salt from seaweed, comprising the step of immersing dried seaweed in water to remove salt.

[0015] More specifically, the present invention provides a method for removing salt from seaweed, characterized in that the steps of soaking dried seaweed in water 30 times its weight and removing the water are repeated three times, or the steps of soaking dried seaweed in water 40 times its weight and removing the water are repeated twice.

[0016] As one specific embodiment, the method for removing salt from seaweed of the present invention can be performed by repeating the steps of immersing dried seaweed in water 30 times its weight and removing the water three times.

[0017] As another specific embodiment, the method for removing salt from seaweed of the present invention can be performed by repeating the steps of immersing dried seaweed in water 40 times its weight and removing the water twice.

[0018] The above dried seaweed refers to seaweed whose moisture content has been reduced to less than 12% by weight by conventional methods such as hot air drying, cold air drying, natural drying, and dehydration drying.

[0019] The above-mentioned immersion can be performed by soaking the dried seaweed in water for 18 to 22 minutes, preferably 19 to 21 minutes, and more preferably 20 minutes. If the above-mentioned immersion is performed for less than 18 minutes, there is a problem that the salt removal from the dried seaweed is not sufficiently performed, resulting in an insufficient effect of reducing the salinity. In addition, if the above-mentioned immersion is performed for more than 22 minutes, the salinity of the seaweed is no longer reduced regardless of the weight of the water at the time of immersion and the number of immersions, making it ineffective in reducing the salinity of the seaweed.

[0020] The above-mentioned desalination of seaweed refers to reducing the salinity of seaweed to less than 0.05%. Seaweed with a salinity of less than 0.05% is suitable as pet food because it does not cause kidney problems when included in pet food.

[0021] After the above immersion is repeated two or three times, the seaweed is dried using a conventional drying method such as hot air drying, cold air drying, natural drying, or dehydration drying, and the seaweed is then ground into powder, and the powdered seaweed can be included in the pet food described below.

[0022] The above-mentioned method for removing salt from seaweed provides a method for optimally removing salt in a process of immersing in water, and the seaweed from which salt has been removed contains less than 0.05% salt, so it can be suitably used as pet food.

[0023] In another aspect, the present invention provides a pet food composition comprising desalinated seaweed.

[0024] The above-mentioned seaweed from which the salt has been removed is seaweed having a salt content of less than 0.05%, preferably seaweed having a salt content of less than 0.05% and a moisture content of less than 12% by weight, and the seaweed is wakame or laver.

[0025] The above seaweed has a higher total phenol content than seaweed leaves and stems, and other seaweeds such as kelp, has an excellent antioxidant effect, and has a high fucoidan content, so it is useful for improving the health of companion animals when included in pet food. Specifically, the above seaweed has a total phenol content that is 41 to 78% higher than seaweed leaves and stems, and kelp, a DPPH radical scavenging activity that is 46 to 118% higher, an ABTS radical scavenging activity that is 98 to 215% higher, and a fucoidan content that is 310 to 1530% higher (see Tables 7 and 8).

[0026] The above seaweed may be contained in an amount of 2.8 to 3.2 wt%, preferably 2.9 to 3.1 wt%, and most preferably 3.0 wt% of the total composition. In the above content range, the fatty acid oxidation rate, total phenol content, and antioxidant activity are the best (see Table 12).

[0027] The above laver has a higher total phenol content than other seaweeds such as seaweed leaves and stems and kelp, has an excellent antioxidant effect, and has a high fucoidan content, so it is useful for improving the health of companion animals when included in pet food. Specifically, the above laver had a total phenol content that was 35 to 79% higher than that of seaweed leaves and stems and kelp, a DPPH radical scavenging activity that was 42 to 118% higher, an ABTS radical scavenging activity that was 98 to 215% higher, and a fucoidan content that was 400 to 1700% higher (see Tables 7 and 8).

[0028] The above-mentioned blue may be contained in an amount of 0.8 to 1.2 wt%, preferably 0.9 to 1.1 wt%, and more preferably 1.0 wt% of the total composition. In the above-mentioned content range, the fatty acid oxidation rate, total phenol content, and antioxidant activity are the best (see Table 12).

[0029] The pet food of the present invention may further include, in addition to the above-mentioned seaweed, chicken, carrot, broccoli, potato powder, soy protein, calcium phosphate, vitamin and mineral mixture, vegetable powder including broccoli and spinach, parsley, organic premix, sweet potato powder, and pumpkin powder. These ingredients may be contained in the total content of the composition as chicken 79 to 81 wt%, carrot 6.8 to 7.4 wt%, broccoli 4.40 to 4.64 wt%, potato powder 5.2 to 5.5 wt%, soy protein 1.3 to 1.6 wt% or 0.3 to 0.6 wt%, calcium phosphate 0.13 to 0.17 wt%, vitamin and mineral combination 0.04 to 0.06 wt%, vegetable powder 0.10 to 0.14 wt%, and parsley, organic premix, sweet potato powder, and pumpkin powder 0.05 to 0.07 wt%, respectively. At this time, the content of the soy protein is 0.3 to 0.6 wt% when the above-mentioned seaweed content is 2.8 to 3.2 wt%, and is 1.3 to 1.6 wt% when the above-mentioned seaweed content is 0.8 to 1.2 wt%.

[0030] When seaweed and other ingredients are contained in the above-mentioned amount, pets can consume them without any aversion, and accordingly, by supplying the pets with fucoidan and phenol ingredients contained in seaweed, they can obtain antioxidant and anticancer effects from these ingredients.

[0031] The above-described pet food composition can be vacuum-packed in a retort pack, thereby facilitating storage and transportation, and can be provided so that the pet can consume it immediately without separate treatment such as heating.

[0032] In another aspect, the present invention provides a method for producing a pet food composition comprising desalinated seaweed.

[0033] More specifically, the method for manufacturing the pet food composition of the present invention is as follows:

[0034] (S10) Step of removing salt by immersing dried seaweed in water;

[0035] (S20) A step of mixing the pet food composition of the present invention;

[0036] (S30) A step of filling the mixture of step (S20) into a retort pack and vacuum packaging it; and

[0037] (S40) A step of heat-treating the retort pack of the above step (S30);

[0038] Hereinafter, a method for manufacturing a pet food composition of the present invention will be described with reference to FIG. 4.

[0039] (S10) Step is a step of removing salt by immersing dried seaweed in water.

[0040] More specifically, it is a step of removing salt from seaweed by repeating the step of soaking dried seaweed in water 30 times its weight and removing the water three times or the step of soaking dried seaweed in water 40 times its weight and removing the water twice.

[0041] The above dried seaweed is seaweed whose moisture content has been reduced to less than 12% by weight by conventional methods such as hot air drying, cold air drying, natural drying, and dehydration drying, and may be wakame or laver.

[0042] The above-mentioned immersion can be performed by soaking the dried seaweed in water for 18 to 22 minutes, preferably 19 to 21 minutes, and more preferably 20 minutes. If the above-mentioned immersion is performed for less than 18 minutes, there is a problem that the salt removal from the dried seaweed is not sufficiently performed, resulting in an insufficient effect of reducing the salinity. In addition, if the above-mentioned immersion is performed for more than 22 minutes, the salinity of the seaweed is no longer reduced regardless of the weight of the water at the time of immersion and the number of immersions, making it ineffective in reducing the salinity of the seaweed.

[0043] The above-mentioned desalination of seaweed refers to reducing the salinity of seaweed to less than 0.05%. Seaweed with a salinity of less than 0.05% is suitable as pet food because it does not cause kidney problems when included in pet food.

[0044] After the above immersion is repeated two or three times, the seaweed is dried using a conventional drying method such as hot air drying, cold air drying, natural drying, or dehydration drying, and the seaweed is then ground into powder, and the powdered seaweed can be included in the pet food described below.

[0045] Step (S20) is a step of mixing the pet food composition of the present invention, and specifically, it is a step of mixing the seaweed powder, chicken, carrot, broccoli, calcium phosphate, vegetable powder, parsley, organic premix, sweet potato powder, and pumpkin powder from step (S10).

[0046] The above mixing can be done as described in the composition described above.

[0047] For example, the mixture may be composed of 2.8 to 3.2 wt% of seaweed, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 0.3 to 0.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral combination, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively.

[0048] For example, the mixture may be composed of 0.8 to 1.2 wt% of seaweed, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 1.3 to 1.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral combination, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively.

[0049] Step (S30) is a step of filling the mixture of step (S20) into a retort pack and vacuum packaging it.

[0050] The above filling can be performed by arbitrarily determining the mixture of the above step (S20), and the vacuum packaging can be performed using a conventional vacuum packaging machine.

[0051] Step (S40) is a step of heat-treating the retort pack of step (S30).

[0052] The above heat treatment is for sterilizing the mixture filled in the retort pack, and can be performed at 100°C or higher, preferably 110°C or higher, and most preferably 120°C for 30 to 40 minutes, preferably 34 to 36 minutes.

[0053] After the above heat treatment, the heated retort pack can be cooled to a temperature of 25°C or lower.

[0054] The pet food composition manufactured by the above-described method can be stored in a sterilized state in a retort pack and can be stored for a long period of time. When the retort pack is opened, the pet food according to the present invention filled in the retort pack can be fed to the pet directly, and the pet can consume it without any resistance.

[0055] The method for removing salt from seaweed according to the present invention is a method for optimally removing salt in a water immersion process, and since it contains less than 0.05% salt, it can be suitably used as a food feed for pets. In addition, since the food feed composition for pets according to the present invention contains seaweed from which salt has been removed, it can be fed to pets by providing the useful components of seaweed without harming the health of the pet due to salt, thereby providing the pets with antioxidant and anticancer effects due to the useful components of seaweed. In addition, since it is food feed despite containing seaweed, it has the effect that the pets can consume it without rejection. In addition, the food feed composition for pets according to the present invention is not only packaged in a retort pack for easy storage and transportation, but also has the advantage of being already cooked, so it can be fed directly to pets without separate heating or cooking.

[0056] Figure 1 is a drawing showing a process for removing salt from dried seaweed according to one embodiment of the present invention.

[0057] Figure 2 is a drawing showing a salt removal process according to the immersion ratio and number of immersions of dried seaweed according to one embodiment of the present invention.

[0058] FIG. 3 is a diagram showing the results of functional group analysis of fucoidan extracted from seaweed powder according to one embodiment of the present invention (A: purified fucoidan (standard material), B: fucoidan extracted from kelp, C: fucoidan extracted from wakame, D: fucoidan extracted from laver).

[0059] Figure 4 is a drawing showing a manufacturing process of a functional edible feed with added seaweed according to one embodiment of the present invention.

[0060] Hereinafter, the present invention will be described in detail using examples to aid understanding. However, the examples according to the present invention may be modified in various ways, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more fully explain the present invention to those of average skill in the art.

[0061] Example 1: Investigation of physicochemical properties of seaweed

[0062] Seaweed is a low-calorie food rich in dietary fiber, such as alginic acid, which can help prevent cardiovascular disease, lower cholesterol, and obesity. Additionally, seaweed is rich in minerals (calcium, iron, iodine, magnesium, phosphorus, potassium, zinc, copper, manganese, selenium, and fluorine), vitamins (A, B1, B2, B9, B12, C, D, E, and K), and polyphenols, which aid in growth and development, prevent osteomalacia and osteoporosis, and have antioxidant properties. With these various beneficial properties, seaweed was investigated to determine its physicochemical properties in order to use it as a raw material in the production of functional pet foods.

[0063] Specifically, the seaweeds used for the production of functional pet foods were purchased and used: kelp (Gijang), wakame (leaves and stems) (Gijang), wakame (Wando), and laver (domestic). The purchased seaweeds were ground to a size of 0.5 mm or less using a hammer mill (LM3100, PerkinElmer, Waltham, MA, USA) and then subjected to physicochemical property analysis.

[0064] All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). Significance (p<0.05) between treatment groups was tested using Duncan's multiple range test.

[0065] 1-1. pH measurement

[0066] 2.5 g of seaweed powder was mixed with 50 mL of distilled water, centrifuged at 3,400 × g for 10 minutes using a centrifuge (Combi R515, Hanil Science Industrial, Incheon, Korea), and the supernatant was separated. The separated supernatant was measured using a pH meter (Accumt ® The pH was measured using a pH meter (AB150, Thermo Fisher Scientific, Singapore).

[0067] 1-2. Salinity of seaweed

[0068] After adding 50 times the amount of distilled water to the seaweed powder, the mixture was mixed at approximately 3,000 rpm for 30 seconds using a vortex mixer (VM-10, Daihan Scientific, Wonju, Korea). The seaweed mixture was centrifuged at 3,400 × g for 10 minutes, and the salinity of the supernatant was measured using a salinity meter (DMT-20, Daeyoon Scale Industry, Seoul, Korea). The salinity (%) of the seaweed powder was calculated by multiplying the measured value by the dilution factor.

[0069] 1-3. Ash content of seaweed

[0070] 3g of seaweed powder was placed in a crucible with a confirmed constant weight and pre-carbonized at 200℃ for 2 hours. Afterwards, it was heated at 600℃ for more than 8 hours, and the sample and crucible were repeatedly heated and cooled until the constant weight was reached. The ash content (%) was calculated using Equation 1 below.

[0071] [Calculation Formula 1]

[0072] Ash content (%) = [(weight of crucible and sample after ash (g) - weight of crucible (g)) / amount of sample collected before ash (g)] × 100

[0073] 1-4. Antioxidant activity of seaweed

[0074] 20 mL of distilled water was added to 1 g of seaweed powder and mixed using a vortex mixer at approximately 3,000 rpm for 30 seconds. The mixed sample was centrifuged (3,400 × g, 10 minutes), and the supernatant was filtered through filter paper (Whatman No. 1, Cytiva, Little Chalfort, UK). The filtrate was used to measure DPPH radical scavenging activity and ABTS radical scavenging activity.

[0075] DPPH radical scavenging activity was measured using a modified version of the method of Blois (1958). 3.5 mL of 60 μM DPPH (2,2-diphenyl-1-picrylhydrazyl) reagent was mixed with 0.5 mL of filtrate, left in a darkroom for 30 minutes, and the absorbance was measured at 517 nm. DPPH radical scavenging activity (%) was calculated using Equation 2 below.

[0076] [Calculation Formula 2]

[0077] DPPH radical scavenging activity (%) = [1 - absorbance of sample / absorbance of blank] × 100

[0078] ABTS radical scavenging activity was measured using a modified method of Re et al. (1999). 7.0 mM ABTS [2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] and 2.45 mM potassium persulfate were mixed in a 1:1 ratio and reacted at room temperature for more than 16 hours to form radicals. On the day of the experiment, the ABTS radical was diluted with ethanol so that the absorbance at 734 nm was 0.700±0.001 and maintained at 30℃ before use. 2.8 mL of ABTS radical was mixed with 0.4 mL of the filtrate and reacted at 30℃ for 30 minutes, and the absorbance was measured at 734 nm. The ABTS radical scavenging activity (%) was expressed using the following calculation formula 3.

[0079] [Calculation Formula 3]

[0080] ABTS radical scavenging activity (%) = [1 - absorbance of sample / absorbance of blank] × 100

[0081] 1-5. Experimental Results

[0082] The experimental results of 1-1 to 1-4 above are shown in [Table 1]. The pH of the seaweeds was the lowest in kelp and wakame seaweed (p<0.05), and the pH of wakame seaweed was the highest at 6.35 (p<0.05). Wakame seaweed also showed the highest salinity and ash content at 10.93% and 31.11%, respectively (p<0.05), but laver showed lower salinity and ash content compared to the other seaweeds (p<0.05). Among the four seaweeds, laver showed the highest DPPH radical scavenging activity and ABTS radical scavenging activity, showing excellent results (p<0.05).

[0083] Type pH Salinity (%) Query (%) DPPH radical scavenging activity (%) ABTS radical scavenging activity (%) Kelp 5.67±0.01 C 7.05±0.17 B 23.17±0.20 B 44.89±0.35 B 53.30±0.18 B Wakame 6.35±0.01 A 10.93±0.29 A 31.11±0.27 A 44.62±0.16 B 52.28±0.14 C Wakame ear 5.70±0.04 C 3.20±0.19 C 21.62±0.08 C 45.32±0.18 B 52.53±0.24 C Blue 6.03±0.00 B 1.00±0.00 D 7.17±0.09 D 48.22±0.20 A 53.86±0.10 A

[0084] All figures represent the mean ± standard error. A-DIndicates significant differences between different letters in the same column (p<0.05).

[0085] Example 2: Investigation of optimal conditions for desalination of dried seaweed.

[0086] 2-1. Investigation of salinity of seaweed immersion solution according to immersion ratio and immersion time

[0087] Seaweed can contain high levels of salt due to its raw material properties and the drying process. High levels of salt can be a burden on pets' kidneys, so using seaweed as a functional ingredient in pet food requires a process to remove salt from the raw material. Therefore, the inventors of the present invention sought to establish optimal conditions for removing salt from dried seaweed.

[0088] In order to confirm the salt removal effect of dried seaweed according to the immersion ratio and immersion time, dried seaweed, which had the highest salt content among the seaweeds purchased as raw materials for functional pet food, was used as the raw material and salt was removed as shown in [Fig. 1]. Water (400, 600, 800, and 1,000 mL, respectively) was added to dried seaweed (20 g) at ratios of 1:20, 1:30, 1:40, and 1:50, respectively, and the seaweed was immersed for 1 hour, and the salt change of the immersion solution over time was measured. After immersing the seaweed for 1 hour at each immersion ratio, the seaweed was taken out and immersed a second time with the same ratio of water mixed. The salt change over time in the second immersion solution was confirmed for 30 minutes, and after the third immersion using the same method, the salt change over time in each immersion solution was investigated for 30 minutes.

[0089] Salinity was measured using a digital salinity meter (DMT-20, Daeyoon Scale Industry, Seoul, Korea) after collecting approximately 10 mL of the immersion liquid used during seaweed immersion. All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). The significance (p<0.05) over time at each immersion ratio was tested using Duncan's multiple range test.

[0090] The changes in salinity according to the immersion ratio and time of the seaweed immersion solution during the first immersion are as shown in [Table 2]. The salinity of the seaweed immersion solution treated with the sample to immersion solution ratio of 1:30 and 1:40 did not increase after 10 minutes (p>0.05) and reached a equilibrium state, but the seaweed immersion solution treated with the ratios of 1:20 and 1:50 did not show a significant change in salinity after 20 minutes of immersion (p>0.05).

[0091] Immersion ratioImmersion time (min)51020304050601:201.08±0.01 C 1.12±0.01 B 1.13±0.00 AB 1.14±0.00 A 1.14±0.00 A 1.14±0.00 A 1.14±0.00 A 1:300.69±0.01 B 0.72±0.01 A 0.72±0.01 A 0.73±0.01 A 0.73±0.01 A 0.73±0.01 A 0.73±0.01 A 1:400.45±0.02 B 0.48±0.03 A 0.49±0.03 A 0.49±0.03 A 0.49±0.03 A0.49±0.03 A 0.49±0.03 A 1:500.37±0.01 C 0.42±0.01 B 0.45±0.00 A 0.45±0.00 A 0.45±0.01 A 0.45±0.01 A 0.45±0.01 A

[0092] All figures represent the mean ± standard error. A-C Indicates significant differences between different letters in the same column (p<0.05).

[0093] The changes in salinity according to the immersion ratio and time of the seaweed immersion liquid during the second immersion are as shown in [Table 3]. The immersion liquids in which the sample and immersion liquid were mixed at ratios of 1:30, 1:40, and 1:50 did not show an increase in salinity after 10 minutes of immersion (p>0.05). On the other hand, in the immersion liquid with a sample and immersion ratio of 1:20, the salinity continuously increased with the passage of time up to 20 minutes of immersion (p<0.05), but after 20 minutes, no change in salinity according to immersion time was observed (p>0.05).

[0094] Immersion ratioImmersion time (min)51020301:200.29±0.00 C 0.33±0.00 B 0.35±0.00 A 0.35±0.00 A 1:300.21±0.00 B 0.22±0.00 AB 0.23±0.00 A 0.23±0.00 A 1:400.12±0.00 B 0.14±0.00 A 0.15±0.00 A 0.15±0.00 A 1:500.07±0.00 B 0.11±0.00 A 0.11±0.00 A 0.11±0.00 A

[0095] All figures represent the mean ± standard error. A-C Indicates significant differences between different letters in the same column (p<0.05).

[0096] The changes in salinity according to the immersion ratio and time of the seaweed immersion solution during the third immersion are as shown in [Table 4]. The seaweed immersion solutions immersed at ratios of 1:20 and 1:30 during the third immersion significantly increased in salinity over time (p<0.05), and maintained a constant salinity after 10 minutes (p>0.05). On the other hand, the seaweed immersion solutions immersed at ratios of 1:40 and 1:50 increased in salinity until 20 minutes (p<0.05), and maintained a constant salinity thereafter (p>0.05).

[0097] Immersion ratioImmersion time (min)51020301:200.13±0.00 B 0.15±0.00 A 0.17±0.00 A 0.17±0.00 A 1:300.05±0.00 B 0.06±0.00 A 0.06±0.00 A 0.07±0.00 A 1:400.04±0.00 B 0.04±0.00 B 0.05±0.01 A 0.05±0.00 A 1:500.01±0.00 C 0.03±0.00 B 0.03±0.00 A 0.03±0.00 A

[0098] All figures represent the mean ± standard error. A-C Indicates significant differences between different letters in the same column (p<0.05).

[0099] The above results indicate that, regardless of the immersion ratio and number of immersions, the salinity of the seaweed immersion solution reaches equilibrium after 20 minutes. Therefore, it was determined that, to most effectively remove salt from dried seaweed, the seaweed must be immersed for at least 20 minutes.

[0100] 2-2. Salinity study of seaweed immersion solution according to immersion ratio and number of immersions

[0101] In the above Example 2-1, the optimal time for removing salt was confirmed, and the following experiment was conducted to confirm the immersion ratio and number of immersions that can most effectively remove salt in seaweed in order to use dried seaweed as a functional pet food raw material.

[0102] Specifically, to remove salt from dried seaweed, water (400, 600, 800, and 1,000 mL, respectively) was added to 20 g of dried seaweed at ratios of 1:20, 1:30, 1:40, and 1:50, and the seaweed was soaked for 20 minutes to remove salt for the first time. After the seaweed was soaked for the first time, the second and third soakings were performed using the same method [Fig. 2].

[0103] Salinity was measured using a digital salinity meter (DMT-20, Daeyoon Instrument Industry Co., Ltd., Seoul, Republic of Korea) after collecting 15 mL of the immersion liquid used during seaweed soaking. The immersion liquid was collected 20 minutes after mixing the seaweed and water. All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). The significance (p<0.05) between treatment groups was tested using Duncan's multiple range test.

[0104] The changes in salinity according to the immersion ratio and number of immersions of the dried seaweed immersion solution are shown in [Table 5]. The salinity of the dried seaweed immersion solution decreased as the immersion ratio increased regardless of the number of immersions (p<0.05). However, the salinity of the dried seaweed immersion solution in the second immersion did not show a significant difference at immersion ratios of 1:40 and 1:50 (p>0.05), and the dried seaweed immersion solution in the third immersion did not show a difference in salinity at immersion ratios of 1:30 or higher (p>0.05). The salinity of the immersion solution decreased significantly as the number of immersions increased at the same immersion ratio (p<0.05), but there was no difference in salinity between the second and third immersion times at immersion ratios of 1:20, 1:40, and 1:50 (p>0.05). Therefore, it was found that salt in the seaweed could be effectively removed when the dried seaweed was soaked twice at an immersion ratio of 1:40 or three times at an immersion ratio of 1:30.

[0105] Number of immersions, immersion ratio 1:20, 1:30, 1:40, 1:50, 1st 7.00±0.46 Ax 4.55±0.30 Bx 3.30±0.22 Cx 1.90±0.20 Dx 2nd 2.00±0.11 Ay 1.30±0.07 By 0.90±0.13 Cy 0.70±0.20 Cy 3rd 1.80±0.08 Ay 0.70±0.07 Bz 0.60±0.08 By 0.60±0.08 By

[0106] All figures represent the mean ± standard error. A-C Indicates a significant difference between different letters in the same row (p<0.05). x-z Indicates significant differences between different letters in the same column (p<0.05).

[0107] Example 3: Investigation of physicochemical properties of desalinated seaweed

[0108] To use seaweed as a functional material in the manufacture of pet food, the physicochemical properties of desalinated kelp, wakame, wakame seaweed, and laver were analyzed.

[0109] Specifically, 500 g of four types of domestically produced dried seaweed (kelp, wakame, wakame ear, and laver) were each soaked in 40 times the volume of water (20 L) for 30 minutes. The soaked seaweed was removed using a sieve and then soaked again in 20 L of water for 30 minutes to remove salt a second time. The seaweeds whose salts were removed were washed once more with running water and then dehydrated using a food dehydrator (OXO Good Grips Salad Spinner, OXO International, Chambersburg, PA, USA). Dehydrated seaweed was spread on a Teflon mesh (hole size: 5 × 5 mm, AM05, Alphaflon, Seoul, Korea), dried in a dryer (EN-FO-392S, Enex Science, Goyang, Korea) set at 60°C for 12 h, and ground into powder using a hammer mill (LM3100, PerkinElmer, Waltham, MA, USA). The seaweed powders were vacuum-packed in polyethylene bags, and then placed in aluminum bags to protect against light. The desalinated seaweed powders were stored at room temperature and analyzed for their physicochemical properties.

[0110] All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). Significance (p<0.05) between treatment groups was tested using Duncan's multiple range test.

[0111] 3-1. pH measurement

[0112] 2 g of seaweed powder was weighed, 100 mL of distilled water was added, mixed, and centrifuged at 3,400 × g for 10 minutes to separate the supernatant. pH meter (Accumt ® The pH of the supernatant was measured using a pH meter (AB150, Thermo Fisher Scientific, Singapore).

[0113] 3-2. Salt measurement

[0114] Salt content was measured using a modified version of the salting method in the Food Code. 2.5 g of seaweed powder was weighed into a salting crucible, incinerated at 600°C for 8 hours, mixed with 25 mL of distilled water, and filtered. 1 mL of 5% K2CrO4 was added to the filtrate, which was titrated with 0.02 N silver nitric acid (AgNO3). Salt content (%) was calculated using Equation 4 below.

[0115] [Calculation Formula 4]

[0116] Salt (%) = [(0.02N AgNO3 consumption (mL) × dilution factor × 0.0011688) / sample amount (g)] × 100

[0117] 3-3. Moisture content

[0118] Moisture content was measured by the 105°C atmospheric pressure drying method according to the AOAC (2016) method.

[0119] 3-4. Ash content

[0120] The ash content was analyzed by direct incineration at 600°C according to the AOAC (2016) method.

[0121] 3-5. Measurement of antioxidant power

[0122] 1 g of seaweed powder was mixed with 50 mL of distilled water using a vortex mixer (VM-10, Daihan Scientific, Wonju, Korea) at approximately 3,000 rpm for 30 s. The mixed sample was centrifuged (3,400 × g, 10 min), and the supernatant was filtered through filter paper (Whatman No. 1, Cytiva, Little Chalfort, UK). The filtrate was used to measure total phenol content, DPPH radical scavenging activity, and ABTS radical scavenging activity.

[0123] Total phenolic contents (TPC) were measured using the method of Singleton et al. (1999). 2 mL of 10% Folin-Ciocalteu's solution was added to 0.4 mL of the filtrate and mixed for 30 s using a vortex mixer. After 5 min, 1.6 mL of 7.5% Na2CO3 was added to the mixture, and the mixture was reacted at room temperature for 120 min. After the reaction was completed, the absorbance of the sample was measured at a wavelength of 765 nm using a spectrophotometer (UV-1800, Shimadzu, Kyoto, Japan). Gallic acid (G7384, Sigma-Aldrich, St. Louis, MO, USA) was used as a standard, and the total polyphenol content was expressed as mg gallic acid equivalent (GAE) per 100 g of the sample.

[0124] DPPH radical scavenging activity and ABTS radical scavenging activity were measured using a method similar to that described in 1-4 above.

[0125] 3-6. Experimental Results

[0126] The pH, moisture, ash, and salt content of the seaweed powders from which the salt was removed are shown in [Table 6]. The pH of the seaweed powder was not different from that of the kelp and seaweed powders (p>0.05), but it was higher than that of the laver powder (p<0.05). The moisture content was the highest in the seaweed powder at 10.50% (p<0.05), and there was no significant difference among the seaweed powders (kelp powder, seaweed powder, and laver powder) except for the seaweed powder (p>0.05). The ash content was significantly lower in the order of kelp (14.70%), seaweed (13.30%), and seaweed (13.01%) (p<0.05), and in particular, the ash content of the laver powder was the lowest at 4.16% (p<0.05). Additionally, the blue powder showed the lowest salt content (0.022%) (p<0.05), and the salt content of the seaweed powders was less than 0.05%.

[0127] Sample pH, moisture (%), ash (%), salt (%), kelp powder 6.60±0.04 B 7.78±0.15 B 14.70±0.09 A 0.042±0.002 AB Wakame powder 6.73±0.04 A 10.50±0.11 A 13.30±0.05 B 0.038±0.003 B Wakame powder 6.67±0.05 AB 7.81±0.07 B 13.01±0.04 C 0.049±0.001 A Blue powder 6.40±0.01 C 7.53±0.06 B 4.16±0.05 D 0.022±0.005 C

[0128] All figures represent the mean ± standard error. A-D Indicates significant differences between different letters in the same column (p<0.05).

[0129] The total phenol content, DPPH and ABTS radical scavenging activity of the seaweed powders from which salt was removed are shown in [Table 7]. The laver powder showed the highest antioxidant activity results (total phenol content, DPPH and ABTS radical scavenging activity) (p<0.05), but the DPPH radical scavenging activity did not show a significant difference from that of the wakame powder (p>0.05). The wakame powder showed higher total phenol content and ABTS radical scavenging activity than the kelp and wakame powders (p<0.05). Meanwhile, the wakame powder showed the lowest antioxidant activity among the four types of seaweed powders (p<0.05).

[0130] Total phenol content of sample (mgGAE / 100g)DPPH radical scavenging activity (%)ABTS radical scavenging activity (%)Kelp powder225.15±4.34 C 29.77±0.49 B 27.49±0.42 C Wakame powder 183.63±4.86 D 20.61±0.59 C 18.10±0.39 D Wakame powder 315.64±3.85 B 43.44±0.40 A 55.70±0.25 B Blue powder 335.85±3.54 A 42.83±1.44 A 57.10±0.17 A

[0131] All figures represent the mean ± standard error. A-D Indicates significant differences between different letters in the same column (p<0.05).

[0132] From the above results, it was determined that all seaweed powders with desalinated salt contained extremely small amounts of salt (less than 0.05%), making them suitable for use as functional ingredients in pet food manufacturing. Among them, laver powder exhibited high antioxidant activity, and kelp powder also showed excellent antioxidant activity, second only to laver powder.

[0133] Example 4: Investigation of a functional substance derived from seaweed (fucoidan)

[0134] Fucoidan, a complex polysaccharide found primarily in brown algae, is a non-toxic ingredient with diverse physiological activities, including anti-inflammatory, immune-boosting, and anticancer effects. Therefore, the inventors of the present invention sought to extract fucoidan from seaweed powder, a functional ingredient used in pet food, and to determine its properties.

[0135] Fucoidan extraction was performed using a slightly modified method of Palanisamy et al. (2017). 20 g of seaweed powder (kelp, wakame, wakame seaweed, and laver) whose salt had been removed in the same manner as in Example 3 was mixed with 85% ethanol at a ratio of 1:20 and stirred at room temperature for 12 hours. After stirring, ethanol was removed using a vacuum filter, and the residue was thoroughly dried at room temperature for more than 12 hours. 200 mL of distilled water was added to 5 g of the dried residue and shaken at 120 rpm for 60 minutes in a 65°C constant-temperature water bath (MaXturdy 45, Daihan Scientific, Wonju, Korea). The shaken solution was centrifuged at 3,400 × g for 10 minutes, and 1% CaCl2 was added to the obtained supernatant and reacted at 3°C ​​for 12 hours to precipitate alginate. The precipitate was removed by centrifugation (3,400×g, 10 min) and ethanol was added to the supernatant to make the concentration 30% and stored at 3℃ for 4 hours. The stored sample solution was centrifuged again to separate the supernatant, and ethanol was added to make the final ethanol concentration of the supernatant 70% and stored at 3℃ for more than 12 hours. After 12 hours, the precipitate (fucoidan) produced was separated from the 70% ethanol solution and lyophilized. The lyophilized fucoidan was pulverized and stored in an airtight container at -24℃ until analysis, and the following experiments were conducted.

[0136] All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). Significance (p<0.05) between treatment groups was tested using Duncan's multiple range test.

[0137] 4-1. Fucoidan extraction yield

[0138] The fucoidan extraction yield (%) was expressed as a percentage by dividing the weight of fucoidan obtained after freeze-drying by the dry weight of the powder treated with 85% ethanol used for fucoidan extraction. The results are shown in [Table 8]. The fucoidan extraction yield was the highest for laver powder at 4.62% (p<0.05), and wakame powder showed a higher fucoidan extraction yield of 3.74% than kelp powder (0.91%) and wakame powder (0.27%) (p<0.05). Wakame powder had the lowest fucoidan extraction yield among all seaweed powders (p<0.05), and wakame powder was excluded from subsequent analysis experiments because it was very inefficient for extracting fucoidan.

[0139] Experimental items Kelp extract Fucoidan Wakame extract Fucoidan Wakame ear extract Fucoidan Seaweed extract Fucoidan Green laver extract Fucoidan Fucoidan Extraction yield (%) 0.91±0.02 C 0.27±0.02 D 3.74±0.09 B 4.62±0.12 A

[0140] All figures represent the mean ± standard error. A-D Indicates significant differences between different letters in the same column (p<0.05).

[0141] 4-2. Total sugar content

[0142] Total sugar content was measured using the phenol-sulfuric acid method of Dubois et al. (1956). The extracted fucoidan powder was diluted to 100 ppm with distilled water. 1 mL of 5% phenol was added to 1 mL of the diluted solution and mixed. 5 mL of sulfuric acid (H2SO4) was added and allowed to stand for 10 minutes. The mixture was then mixed using a vortex mixer, reacted at 25°C for 20 minutes, and the absorbance was measured at 490 nm. Glucose (G7021) from Sigma-Aldrich was used as the standard.

[0143] The results are shown in [Table 9] below. The total sugar content of fucoidan extracted from seaweed was the lowest in kelp-extracted fucoidan at 19.59% (p<0.05), while that of fucoidan extracted from laver was the highest at 31.91% (p<0.05).

[0144] Total sugar content (%), DPPH radical scavenging activity (%), ABTS radical scavenging activity (%), kelp extract fucoidan 19.59±0.28 C 31.67±0.31 B 75.88±0.21 C Fucoidan extracted from seaweed 26.31±0.27 B 31.40±0.64 B 82.02±0.25 A Fucoidan extracted from blue seaweed 31.91±0.08 A 39.21±0.33 A 76.93±0.20 B

[0145] All figures represent the mean ± standard error. A-D Indicates significant differences between different letters in the same column (p<0.05).

[0146] 4-3. Antioxidant activity

[0147] To measure antioxidant activity, the fucoidan extract powder was diluted 1000 times with distilled water, and then DPPH radical scavenging activity and ABTS radical scavenging activity were measured using a method similar to that described in Example 1-4.

[0148] The results are as shown in [Table 9] above. Fucoidan extracted from laver had the highest DPPH radical scavenging activity (p<0.05), and there was no difference in DPPH radical scavenging activity between fucoidans extracted from kelp and seaweed (p>0.05). However, fucoidan extracted from laver was superior to fucoidans extracted from kelp and seaweed (p<0.05).

[0149] 4-4. Functional group analysis

[0150] Functional group analysis was performed using a modified version of the method of Husni et al. (2022). Fucoidan extracted from seaweed was mixed with potassium bromide (KBr) at a ratio of 1:100 and homogenized. The homogenized sample was compressed at 7,000 Pa to produce KBr pellets. The KBr pellets were analyzed using a Fourier Transform Infrared (FT-IR) spectrophotometer (FT-IR6100, Jasco, Tokyo, Japan) at a wavelength of 4,000–400 cm -1 The transmittance spectrum was measured over a wavelength range of . Purified fucoidan was used as a standard material.

[0151] The result is as shown in [Figure 3]. Approximately 3,425 cm -1 and 2,929cm -1 In the spectral range, the absorption areas are OH and CH stretching of carbohydrates, respectively, and are around 1650 cm -1 The absorption area in the spectrum represents C=O stretching.

[0152] In particular, key information about the location of the sulfate group is in the spectrum 1500-700 cm -1 Appears in the range of approximately 1,420, 1,255, and 1,055 cm -1 The absorption area in is S=O stretching, which is a characteristic of fucoidan compounds, and is 842 cm -1 The absorption area observed in the vicinity is known to represent sulfated polysaccharides (COS). All fucoidans isolated from seaweeds exhibited the functional group characteristics of fucoidan compounds, and among them, fucoidan extracted from Mekabu showed a very similar FT-IR spectral pattern to the reference material. These results indicate that fucoidan extracted from Mekabu is structurally most similar to purified fucoidan.

[0153] The results of the above experiments are summarized as follows.

[0154] As a result of the quality characteristics of seaweed, all seaweed powders with salt removed had a salt content of less than 0.05%, making them suitable as raw materials for pet food. Among the seaweed powders, laver powder had very high antioxidant activity (total phenol content, DPPH and ABTS radial scavenging activity). Although the wakame seaweed powder had lower antioxidant activity and fucoidan extraction yield than laver powder, it was superior to other seaweed powders. Although the extraction yield of fucoidan extracted from laver seaweed powder was lower than that of laver powder, it was structurally most similar to purified fucoidan. Therefore, laver seaweed or laver powder were judged to be the most suitable materials for manufacturing seaweed-based functional pet food.

[0155] Example 5: Manufacturing of pet food with added seaweed and investigation of its quality characteristics.

[0156] The effects of the type and amount of seaweed added on the physicochemical properties of pet food were investigated using two types of seaweed powder (seaweed and laver) selected through the above examples, and the mixing ratio of seaweed-based functional pet food was determined.

[0157] Specifically, to analyze the quality characteristics of seaweed-based functional pet food, retort-processed pet food was manufactured by adding 0%, 1%, or 3% of seaweed and laver powder, respectively [Fig. 4]. The seaweed-based functional pet food was prepared by mixing the raw materials according to [Table 10] for 30 minutes using a silent cutter. The mixed raw materials were 49×102×18 mm in size. 3 The raw materials were molded to size and vacuum-packed in transparent retort packs (polyester / nylon / polypropylene). The packaged raw materials were heat-treated at 120°C for 35 minutes and cooled for 20 minutes to ensure that the final product temperature was below 25°C. The manufactured edible feed was stored at room temperature and its physicochemical properties were analyzed.

[0158] All experiments were repeated three times, and the measurement results were statistically processed using the GLM (Generalized Linear Model) process of the SAS program (2012, version 9.4). Significance (p<0.05) between treatment groups was tested using Duncan's multiple range test.

[0159] Ingredients (%) Seaweed spray added Seaweed powder added Seaweed powder added 1.0% 3.0% 1.0% 3.0% Chicken 80.0280.0280.0280.0280.0280.02 Carrot 7.107.107.107.107.10 Broccoli 4.5 24.5 24.5 24.5 24.5 24.52 Potato powder 5.8 5 5.35 4.35 5.35 4.35 Soy protein 1.9 5 1.45 0.45 1.45 0.45 Seaweed powder 0.001 003 000 000 000 Seaweed powder 0.000 000 001 003 00 Calcium phosphate 0.15 0.15 0.15 0.15 Vitamin Mineral Mixture 0.050.050.050.050.050.05 Broccoli Powder 0.060.060.060.060.06 Spinach Powder 0.060.060.060.060.06 Parsley 0.060.060.060.060.06 Organic Premix 0.060.060.060.060.06 Sweet Potato Powder 0.060.060.060.060.06 Pumpkin Powder 0.060.060.060.060.06 Total 100.00100.00100.00100.00

[0160] 5-1. Color

[0161] The color of the sample was measured using a chroma meter (CR-400, Konica Minolta Sensing, Osaka, Japan). The light source used for chromaticity measurement was D 65 , and was used after correction using a standard white board (L* value +94.87, a* value -0.36, b* value +3.85).

[0162] The results are shown in [Table 11]. In all color indices (CIE L*, a*, and b*), the edible feed with added seaweed showed lower results than the feed without added seaweed (p<0.05), and as the amount of added seaweed increased, the CIE L*, a*, and b* of the feed significantly decreased (p<0.05). When the amount of added seaweed was the same, the feed containing seaweed powder showed higher CIE L* and b* than the feed containing laver powder (p<0.05), and in particular, the CIE a* was lower in the feed with added laver powder than in the feed with added laver powder regardless of the amount of added seaweed (p<0.05).

[0163] Test Items: No seaweed powder, 1% seaweed, 3% seaweed, 1% seaweed, 3% seaweed, CIE L*64.40±0.20 A 56.93±0.24 B 54.08±0.18 D 56.16±0.19 C 49.50±0.22 E CIE a*0.28±0.08 A -0.11±0.15 B -1.57±0.11 C -2.30±0.11 D -4.57±0.12 E CIE b*28.37±0.24 A 27.63±0.26 B 26.70±0.16 C 26.46±0.14 C 24.01±0.21 D

[0164] All figures represent the mean ± standard error. A-E Indicates significant differences between different letters in the same column (p<0.05).

[0165] 5-2. pH

[0166] The pH of the sample was measured by adding 45 mL of distilled water to 5 g of the sample and homogenizing it, then measuring it with a pH meter (Accumt ⓡThe pH was measured using AB150, Thermo Fisher Scientific, Singapore. The results are shown in [Table 12] below. The addition of seaweed increased the pH of the sorghum feed (p<0.05), and the pH of the feed containing 3% seaweed powder was 6.76, the highest among all sorghum feeds (p<0.05).

[0167] 5-3. Shear force

[0168] The shear force was measured using a Texture analyzer (TA.XTplusC, Stable micro system, Vienna Court, UK) equipped with a Warner-Bratzler blade after cutting each sample into a size of 2.0 cm in height and 1.5 cm in width. The test speed was 5 mm / s. The results are shown in [Table 12] below. The feed with added seaweed had a lower shear force than the feed without added seaweed (p<0.05). In particular, laver powder reduced the shear force of the sashimi feed more significantly than the addition of kelp powder, regardless of the amount added (p<0.05).

[0169] 5-4. Fatty acid oxidation

[0170] Thiobarbicturic acid reactive substance (TBARS) was analyzed according to the method of Tarladgis et al. (1960). Malonaldehyde (MDA) generated by lipid oxidation was reacted with a 0.02 M 2-thiobarbituric acid (TBA) solution, and the absorbance (A) was measured at a wavelength of 538 nm. 538nm ) was measured and expressed as mg of MDA per kg of sample using the calculation formula 4 below.

[0171] [Calculation Formula 4]

[0172] Fatty acid content (mg MDA / kg) = A 538nm × 7.8

[0173] The results are shown in [Table 12] below. The fish feed manufactured by adding seaweed had lower fatty acid oxidation rate (TBASRS) than the feed without seaweed (p<0.05), and laver powder was more effective than kelp powder in reducing fatty acid oxidation rate of the fish feed (p<0.05).

[0174] 5-5. Antioxidant activity

[0175] To measure the antioxidant activity of seaweed-supplemented fish feed, antioxidant components were extracted using a modified method from Mancini et al. (2015). 10 mL of ethanol was added to 5 g of the sample, which was homogenized at 10,000 rpm for 30 s. The homogenized sample was centrifuged (15,000 × g, 10 min), and the supernatant was filtered through filter paper (Whatman No. 1, Cytiva, Little Chalfort, UK). The filtrate was used to measure total phenol content, DPPH radical scavenging activity, and ABTS radical scavenging activity.

[0176] Total phenol content was measured using the same method as described in Examples 3-5 above, and DPPH radical scavenging activity and ABTS radical scavenging activity were measured using a method similar to the method described in Examples 1-4 above.

[0177] The results are shown in [Table 12] below. Total phenol content was higher in feeds containing seaweed than in those without seaweed (p<0.05), and was highest in feeds containing laver (p<0.05). However, in the same seaweed-containing feeds, there was no difference in total phenol content according to the amount of seaweed added (p>0.05).

[0178] As with the results for total phenol content, all seaweed-added croaker feeds showed higher DPPH radical and ABTS radical scavenging activities than feeds without seaweed (p<0.05). The DPPH radical scavenging activity of croaker feeds with added seaweed powder increased as the amount increased (p<0.05), but there was no difference in DPPH radical scavenging activity according to the amount of added seaweed in the croaker feeds with added laver powder (p>0.05). In ABTS radical scavenging activity, the croaker feed with added 1% laver powder did not show a significant difference from the other seaweed-added treatments (p>0.05), but when 3% laver powder was added, the ABTS radical scavenging activity was lower than that of the feed with added 3% laver powder (p<0.05).

[0179] Experimental items: No seaweed powder, 1% seaweed, 3% laver, 1% laver, 3% pH 6.38±0.03 C 6.69±0.01 B 6.76±0.01 A 6.72±0.01 B 6.68±0.01 B Shear force (N) 7.95±0.39 A 4.78±0.23 B 4.56±0.16 B 3.28±0.10 C 3.32±0.09 C TBARS (mg MDA / kg) 2.17±0.03 A 1.28±0.06 B 1.34±0.05 B 1.06±0.03 C 1.07±0.03 C Total phenol content (mg GAE / 100g) 43.02±0.67 C 61.07±0.48 B 61.60±0.65 B 63.51±0.60 A 63.92±0.75 A DPPH radical scavenging activity (%) 6.29±0.61 C 47.49±0.66 B 51.88±1.52A 49.67±0.79 AB 49.61±0.90 AB ABTS radical scavenging activity (%) 87.60±0.75 C 89.11±0.40 AB 90.29±0.18 A 88.99±0.34 AB 88.51±0.22 B

[0180] All figures represent the mean ± standard error. A-C Indicates significant differences between different letters in the same column (p<0.05).

[0181] The results of the above experiment are as follows.

[0182] To develop a functional pet food based on seaweed, the physicochemical properties of the crosby feed manufactured by adding 0%, 1%, or 3% of Wakame and Laver were analyzed. The addition of seaweed lowered the shear force of the crosby feed, providing a soft texture, and reduced the total phenol content and radical scavenging activity along with the fatty acid oxidation level (TBARS). The feed with added laver powder had a lower fatty acid oxidation level and a higher total phenol content than the feed with added Wakame powder alone. However, the crosby feed with 3% Wakame powder and 1% Laver powder showed the best antioxidant activity in terms of radical scavenging activity.

[0183] Therefore, when manufacturing a seaweed-based functional pet food, it is most desirable to add 3% of seaweed powder or 1% of laver powder, and it was judged that the disadvantages of each powder can be supplemented and the advantages can be maximized through the addition of a mixture of seaweed powders.

Claims

1. A method for removing salt from seaweed, characterized in that the step of soaking dried seaweed in water 30 times its weight for 18 to 22 minutes and removing the water is repeated three times, or the step of soaking dried seaweed in water 40 times its weight for 18 to 22 minutes and removing the water is repeated twice.

2. A method for removing salt from seaweed in the first paragraph, characterized in that the salt content of the seaweed is contained to be less than 0.05%.

3. A pet food composition comprising seaweed from which salt has been removed, wherein the seaweed from which salt has been removed is laver or wakame having a salt content of less than 0.05% and a moisture content of less than 12% by weight. A pet food composition characterized in that the seaweed from which the salt has been removed is prepared by repeating the steps of soaking the dried seaweed in water 30 times its weight for 18 to 22 minutes and removing the water three times, or by repeating the steps of soaking the dried seaweed in water 40 times its weight for 18 to 22 minutes and removing the water twice.

4. A pet food composition, characterized in that in the third paragraph, the seaweed or laver is contained in an amount of 2.8 to 3.2 wt% or 0.8 to 1.2 wt%, respectively, of the total composition.

5. In the third paragraph, the pet food composition is characterized in that it comprises 2.8 to 3.2 wt% of seaweed, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 0.3 to 0.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral combination, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively.

6. In the third paragraph, the pet food composition is characterized in that it comprises 0.8 to 1.2 wt% of laver, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 1.3 to 1.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral combination, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively. 7.(S10) Step of removing salt by immersing dried seaweed in water; (S20) A step of mixing the seaweed powder, chicken, carrot, broccoli, calcium phosphate, vegetable powder, parsley, organic premix, sweet potato powder, and pumpkin powder from which salt has been removed in the step (S10); (S30) A step of filling the mixture of step (S20) into a retort pack and vacuum packaging it; and (S40) A method for producing a pet food composition, comprising a step of heat-treating the retort pack of the step (S30).

8. In the 7th paragraph, the step (S20) is a method for producing a pet food composition characterized in that the seaweed from which salt has been removed is mixed with 2.8 to 3.2 wt% of seaweed, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 0.3 to 0.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral mixture, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively.

9. In the 7th paragraph, the step (S20) is a method for producing a pet food composition characterized in that the seaweed from which salt has been removed is mixed with 0.8 to 1.2 wt% of laver, 79 to 81 wt% of chicken, 6.8 to 7.4 wt% of carrot, 4.40 to 4.64 wt% of broccoli, 5.2 to 5.5 wt% of potato powder, 1.3 to 1.6 wt% of soy protein, 0.13 to 0.17 wt% of calcium phosphate, 0.04 to 0.06 wt% of vitamin and mineral mixture, 0.10 to 0.14 wt% of vegetable powder, and 0.05 to 0.07 wt% of parsley, organic premix, sweet potato powder, and pumpkin powder, respectively.

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