KR100847354B1 - A method for breeding animal using by the leaf of Pinus Koraiensis and the functional animal meat products using thereby - Google Patents

Abstract

The present invention has an antioxidant activity ( Pinus pine needles) Koraiensis ) relates to a method of animal breeding and functional products produced using the same, the pine needles of the present invention have an antioxidant effect and give benefits of improving meat quality, reducing odor and preventing various adult diseases. Can contribute to increased farm income.

 Pine nuts, breeding methods, feed, animals, antioxidants.

Description

A method for breeding animal using by the leaf of Pinus Koraiensis and the functional animal meat products using Thus}

1 is a diagram showing the antioxidant activity of the pine leaf extract against the peroxyl radical,

2 is a diagram showing the antioxidant activity of the leek extract against the peroxyl radical,

3 is a diagram showing the antioxidant activity of glutathione on the peroxyl radical,

Figure 4 is a diagram showing the antioxidant activity of the pine leaf extract against hydroxy radicals,

5 is a diagram showing the antioxidant activity of leek extract against hydroxyl radicals,

6 is a diagram showing the antioxidant activity of glutathione to hydroxyl radicals,

7 is a diagram showing the antioxidant activity of the pine leaf extract against peroxide nitrate,

8 is a diagram showing the antioxidant activity of leek extract against peroxide nitrate,

9 is a diagram showing the antioxidant activity of glutathione for peroxide nitrate.

 The present invention relates to an animal breeding method using a pine leaf showing an antioxidant activity and a functional product produced using the same.

Oxygen-related oxidants in the human body are called reactive oxygen species (ROS) and reactive nitrogen species (RNS) .These types of ROS and RNS are superoxide and hydroxyl. Free radicals, such as peroxyl, alkoxyl, hydroperoxyl and hydrogen peroxide, hypochlorous acid, ozone There are non free radicals, such as singlet oxygen, peroxinitrite, and the like. Among these, reactive, causing direct toxicity is hydroxyl, peroxyl, phenoxynitrite, etc. (Bayir H. et al., Crit Care Med ., 33 (12 Suppl), ppS498-501, 2005; Niles et al., Nitric Oxide , 14 , pp 109-121, 2006).

Oxidizing substances are produced by the redox reactions of various organisms, which can lead to the deterioration of edible fats or oxidative damage to various biological substances (lipids, proteins, nucleic acids, carbohydrates). May be induced (Yen GC et al., J. agric .. Food Chem . , 43 , pp27-32, 1995). Unsaturated fatty acids of phospholipids, which are constituents of biological membranes, initiate peroxidation reactions by free radicals such as reactive oxygen species and proceed in a chain. Therefore, free radical peroxidation not only promotes cell membrane permeability but also results in overall cytotoxicity, which induces aging or pathology of various diseases and is involved in carcinogenesis. Radical action has a great effect on the progression of various chronic diseases such as atopic diseases such as oxidative stress, cancer, hypertension, myocardial infarction, arteriosclerosis, rheumatism, cataracts and Parkinson's disease (De Souza LC. Et al . , Bioorg. Med. Cehm. Lett ., 14, pp5859-5861, 2004), may act as a factor that affects the immune system function (Pike J. et al., Int . J. Vitam. Nutr. Res., 65 , pp 117-120, 1995).

When a living organism is repeatedly exposed to oxidative stress, the cell protects the living body by increasing the production of enzymes or substances having antioxidant activity in response to this. The protective mechanism of these organisms is called an antioxidant defense system and is largely divided into antioxidant enzymes and low molecular weight antioxidants. Antioxidant enzymes and small molecule antioxidants work together to detoxify the oxidizing substances produced in living organisms. In particular, glutathione is a representative low-molecular-weight antioxidant, which not only directly reduces oxidative substances but also acts as a substrate for glutathione S-transferase and glutathione peroxidase. Detoxify (Estrela et al., Crit . Rev. Clin . Lab. Sci ., 43 (2): pp143-181, 2006).

Recently, various attempts have been made to measure the total antioxidant activity in natural product extracts (Prior RL. Et al., J. agric .. Food Chem . , 53 , pp4290-4302, 2005). Determination of total antioxidant activity of samples is effective in determining resistance to reactive oxygen species and several methods have been developed for this purpose (Prior RL. Et al., J. agric .. Food Chem . , 53 , pp 4290-4302, 2005). In the mid-1980s, the experimental system containing free radicals and antioxidants measured the time required for maximum oxygen consumption, and compared the results with the time measured with Troox, the water-soluble vitamin E derivative. trapping antioxidant parameter) method was developed (DeLange and Glazer, Anal Biochem . , 28 , pp 300-306. 1989). Subsequently, a method of measuring antioxidant activity by reducing fluorescence caused by chemical damage to radicals has been proposed (Tanaka et al., J. Am. Chem . Soc ., 123 , pp 2530-2536, 2001). Fluorescence reduction rate of the fluorescent material by the radical is constant, but in the presence of antioxidants, a temporary delay is observed and then the fluorescence decreases rapidly. Therefore, the ORAC (Oxygen Radical Absorbance Capacity) method, which measures the difference between AUC of the sample and the control, and the method of measuring the protection time, or lag-phase, of phosphors in the presence of antioxidants, have been developed (Ghiselli et al. , Free Radical Biol . Med ., 18 , pp 29-36, 1995). However, all of these methods have limitations in evaluating antioxidant activity in vivo by using artificial radicals rather than in vivo radicals (Prior RL. Et al., J. agric .. Food Chem . , 53 , pp 4290-4302, 2005).

Regoli and Winston have developed a method for measuring the antioxidant activity of biological tissues and natural products using α-keto-γ-methiolbutyric acid (KMBA), which reacts with oxidizing substances in vivo and is oxidized to ethylene. (Winston et al., Free Radical Biol. Med ., Feb., 24 (3), pp 480-493, 1998; Regoli and Winston, Toxicol . Appl . Pharmacol., Apr. 15, 156 (2) , pp 96-105, 1999). Ethylene can be quantified by gas chromatography by taking air samples in a closed reaction vessel. The advantage of this method is its utility. KMBA reacts with various oxidizing substances such as hydroxyl radical, peroxyl radical, peroxynitrite and hypochlorous acid to produce ethylene. The generation of ethylene is reduced in this high tissue or blood sample. In this method, the concentration of ethylene produced by the reaction of KMBA with hydroxyl radical, peroxyl radical, peroxynitrite, etc. is measured according to the change of reaction time, and then AUC (area under the curve) and TOSC is calculated from the ratio of AUC in the control group and the administration group. Therefore, it is a highly reproducible and accurate method that can correct the error of the method measured only at a single reaction time. Since the TOSC value measured from the sample is compared with that in the control group, this value is theoretically not affected by the sensitivity of the instrument, the reagent used, or other reaction conditions. Moreover, it is a highly applicable method of organisms by evaluating the antioxidant activity against oxidizing substances actually produced in living bodies (Ghiselli A. et al., Free Radical Biol . Med ., 18 , pp29-36, 1995).

A cooperative defense system that protects the body from free radicals contains oxidative nutrients and enzymes (Halliwell B. et al., Annu . Rev. Nutr . , 16 , pp 33-50, 1996). One effective way to protect the body from oxidative stress is to increase antioxidant levels (De Souza LC. Et al., Bioorg . Med . Cehm . Lett. , 14 , pp5859-5861, 2004). Therefore, the antioxidant evaluation of alternative substances to prevent from oxidative damage is very active.

Antioxidants are used for the purpose of minimizing the loss of certain vitamins and essential amino acids, or delaying or preventing the dispersal of maintenance products by reacting with free radicals rather than removing or absorbing oxygen. Synthetic antioxidants commonly used in foods and pharmaceuticals include butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl galate (PG) and tertiary butyl hydro Quinone (TBHQ, Teritiarybutyl hydroquinone) and the like, but when administered to high concentrations in experimental animals are known to cause liver hypertrophy or carcinogenicity. In particular, butylated hydroxytoluene has been shown to increase microsomal enzyme activity in the liver of laboratory animals through several studies, and controversy has been raised about the safety of these phenolic synthetic antioxidants. Usage is legally regulated (Brannen et al ., J. Amer . Oil Chem . Soc . , 52 , pp59-63, 1975; Ito N. et al., J. Natl . Cancer Inst . , 70 , p343, 1983; Chan KM. Et al., J. Food Sci . , 58 , pp 1-4, 1993).

Accordingly, many studies have been made in the hope of developing natural antioxidants of safe and economical plant origin with high antioxidant effects (Larson RA. Et al., Phytochemistry , 27 , pp969-978, 1988). Recently, as research on natural products has been actively conducted, secondary metabolites contained in natural products have become a major concern as physiologically active substances, and in particular, studies on antioxidants have been actively conducted (中 谷 延 二, 日本 食品). Industrial Engineering, 37 , p569, 1990). Bioactive substances are high value-added substances that show remarkable activity even in very small amounts, and many kinds are usefully used, and new substances are being actively researched (條 原 和 毅, 食品 と 開發 , 27 , p29, 1992; Kang SS. Et al., Natural product science.Seoul Univ. Publishers , Seoul, p71, 1988).

Natural antioxidants known to date include tocopherols, flavonoids, gossypol, sesamol, oryzanol and vitamin C (Huson B. et al ., Food Chem . , 19 , pp 537-541, 1987; Frankel, EN et al., Food Chem . , 57 , p51, 1996; Giese J. et al., Food Technol. , 5 , pp73-81, 1996; Pszcczola DE. Et al., Food Tech. , 55 , pp 51-59, 2001). Among them, tocopherol and L-ascorbic acid are preferred as natural antioxidants. Among them, tocopherol has high safety but low ability to inhibit oxidation reaction (Halliwell B. et al., FASEB) . J. , 2 , pp2867-2870, 1988) is expensive.

Pine tree is a kind of conifer belonging to the pine family. Its constituents include estolide lead, which is composed mainly of essential oils, flavonoids, ascorbic acid and uniper acid, and essential oils on branches, finitol on necks, Pinosylbin, pinosylbin-methyl ester, flavonoids such as chrysine, pinosembrine, pinobanksin, kryptostrobin, seeds are composed of oil, protein, ash, carotene, vitamin B ₂ and protein is arginine, histidine Amino acids such as lysine, tyrosine, leucine and glutamic acid are contained, and tannins are contained in the shell.

The pharmacological effect of the fruit haesongja menopause, stomach and large intestine, and acts to protect the core and heart, lungs, eliminate wind and make urine well. It has nutrition and prevents atherosclerosis, and is used for elderly constipation, systemic weakness, vertigo, dry cough, and hemorrhage. It is also effective for atopic dermatitis, boils, corn, neuralgia, and obesity. , North Africa, 2003).

The leaves are used as antiseptic, urinary medicine, cough medicine and sweat medicine, and eggplants are used for rheumatism, gout, and back pain (interested by medicinal herb, NK Encyclopedia, 1984).

However, none of the literature teaches or mentions the antioxidant activity of pine needles.

The present inventors have completed the present invention by confirming the antioxidant activity of the pine needles through experiments using the TOSC assay.

An object of the present invention is a pine needle having an antioxidant activity ( Pinus Koraiensis ) to provide a method of animal breeding and functional products produced using the same.

In order to achieve the above object, the present invention, the pine needle having an antioxidant activity (Pinus KoraiensisAnimal breeding method using) and provides a functional product produced using the same.

Specifically, the present invention comprises the first step of collecting and grinding the pine needles; A second step of blending the pine needles into the feed using 5 to 10% by weight of the feed weight; It provides a feed method for the animal to the vegetarian diet to provide a method of animal breeding using a pine leaf having an antioxidant activity comprising a third step of producing a functional product.

In the second step, the constituent blending ratio of the feed formulated with pine needles is preferably a weight ratio (w / w) of feed: pine needles (15-25: 0.1-10), more preferably (15-20: 1 It is preferably formulated in a weight ratio (w / w) of -5), and also with respect to the total weight of the feed of the present invention, the pine leaves are 0.5 to 20%, more preferably 1 to 15%, most preferably 5 to 10 It is characterized by containing% as an active ingredient.

Hereinafter, the present invention will be described in detail.

Animal breeding method using the pine needles having an antioxidant activity of the present invention can be provided as follows.

Specifically, the first step of collecting the leaves of the pine tree growing in the wild domestic, and then crushing with a grinder; The pine needles prepared in the first step are used in combination with commercial animal feeds, and the amount of pine tree leaves is used in the composition ratio of the feed to the feed usage: weight ratio of feed: pine needles (15-25: 0.1-10) (w / w), more preferably in a weight ratio (w / w) of (15 to 20: 1 to 5), and also to the total weight of the feed of the present invention, the pine leaf is 0.5 to 20%, more preferably Preferably a second step of formulating the feed using 1 to 15%, most preferably 5 to 10%; It provides an animal breeding method using the pine tree leaf having the antioxidant activity of the present invention through the process of the third step to free the vegetarian compound of the pine leaf compounded compound formulated in the second step.

The animals include mammals such as horses, pigs, cattle, sheep, goats, camels and limas; Red sea bream, sea bream, bastard halibut, flounder, defense, defense fry, amberjack, tuna, yellow salmon, sweetfish, salmon, trout, tiger puffer fish, eel, loach, catfish, carp and goldfish Fish such as; Shellfish such as abalone, turban cells, scallops and oysters; Crustaceans such as prawns, black tiger prawns, yellow sea prawns and blue crab; And farmed animals of poultry such as pheasants, chickens, ducks, turkeys and ostriches and pets such as dogs and cats. The present invention can be widely applied to terrestrial animals and aquatic animals.

In addition, the present invention provides a functional product using a pine leaf having an antioxidant activity produced by the animal breeding method.

Functional products produced using the antioxidant activity of the present invention is characterized in that the raw meat and by-products thereof or processed meat and by-products thereof.

The raw meat and by-products thereof include pheasant meat, chicken meat, pork meat, horse meat, beef meat, lamb meat, duck meat, artificial meat, turkey meat, rabbit meat, dried eggs, eggs, frozen eggs, quail eggs, powdered eggs, duck eggs, and liver. It is characterized by.

The processed meat and by-products thereof are made of liver (Pate), liver paste, meat extract, meat jelly, pork cutlet, Rennet, bacon, bouillon, bouillon, broth, black pudding (Blood Sausage), Beefsteak, Charcuterie, Sausage, Ensign, Edible Bone Marrow, Edible Gelatin, Beef Jerky, Canned Meat, Canned Meat, Baby Food Meat, Ham, Hamburger Meat, Mashed Sausage, Sundae , Giblets and sweet and sour pork.

Hereinafter, the present invention will be described in detail by Examples and Experimental Examples.

However, the following Examples and Experimental Examples are only illustrative of the present invention, and the content of the present invention is not limited by the following Examples and Examples.

Example  1. Animal Breeding Method Using Pine Tree Leaves

The method of animal breeding using pine needles is to collect the leaves of pine trees native to wild domestic, then grind them with a grinder and mix them with 5% by weight of the animal feed on the market. Seven weeks old Duroc, Landrace and Large white species pigs were fed for 4 weeks. The size of the piglet was 3 × 2.5m, the feed was free vegetarian, the water was free to eat using an automatic water dispenser, and the antioxidant experiment of the present invention in the plasma obtained from the functional pigs bred by the above method Used for.

Experimental Example  1. Measurement of Antioxidant Activity

1-1. Sample Preparation

1-1-1. Preparation of Pine Leaf Extract

Pine tree ( Pinus) Koraiensis ) leaves 80.45 g (Gongju-si, Chungnam), cut into 2-4 cm, and extracted by sonication three times at room temperature for 1 hour in 500 ml of 80% methanol.

The extract was concentrated at 45 ° C. for 2 hours using an evaporator and then lyophilized for 3 days. After lyophilization, the dry weight was finally 5.31 g, which was used as a sample of the antioxidant experiment of the present invention.

1-1-2. Preparation of Leek Extract

Domestic leek ( Allium Tuberosum , Walmart, Yuseong-gu , Daejeon Metropolitan City, 80.45 g were taken and cut into 2 ~ 4 cm and extracted by sonication three times at room temperature for 1 hour in 500 ml of 80% methanol.

The extract was concentrated at 45 ° C. for 2 hours using an evaporator and then lyophilized for 3 days. After lyophilization, the dry weight was finally 5.24 g, which was used as a comparative sample of the antioxidant experiment of the present invention.

1-1-3. Preparation of Laboratory Animals

Blood was taken from the jugular vein of the pig to obtain plasma in the pig of Example 1, and then centrifuged at 5,000 × g for 1 minute at room temperature to obtain plasma and stored at −70 ° C. until analysis. To measure the antioxidant activity, the plasma was used diluted 200 or 250 times with distilled water for injection.

1-2. Experiment method

In order to measure the antioxidant activity of the pine leaf extract was carried out a radical scavenging experiment as follows using a total oxygen scavenging capacity (TOSC) assay.

TOSC assays are proposed by Winston et al., Free Radic . Biol . Med ., Feb, 24 (3), pp480-493, 1998 and modified by the same authors (Regoli and Winston, Toxicol . Appl . Pharmacol ., Apr 15, 156 (2) , pp96-105, 1999). Peroxyl radical (Peroxyl radical) is generated by sikyeotgo (thermal homolysis) a free-radical reaction at 35 ℃ the ABAP (2,2'-azobisamidinopropane) (Winston et al., Free. Radic. Biol Med ., Feb, 24 (3), pp 480-493, 1998), hydroxyl radicals are the Fenton reaction using Fe and ascorbate (Fenton reaction; Winston and Cederbaum, Alcholol). Clin . Exp . Res., 9 (2) , pp95-102, 1985), peroxynitrite was generated through spontaneous decomposition of SIN-1. Each reactive oxygen species generated reacts with α-keto-γ-methiolbutyric acid (KMBA) to generate ethylene gas, which does not show any temperature-dependent difference. (Winston et al., Free Radic . Biol . Med ., 24 (3) , pp480-493, 1998; Regoli and Winston, Toxicol . Appl . Pharmacol . , 156 (2) , pp96-105, 1999 ).

The reaction proceeded by putting the final volume of 1 ml of the reaction solution into a sealed 10 ml container with rubber septum. Ethylene gas generation detection takes a certain volume of air in the head space of the reaction vessel, and is equipped with a gas chromatography equipped with a Poropack N column and a FID (flame ionization detector). Was analyzed.

As a result of the experiment, the area under the kinetic curve (AUC) was obtained by integrating a graph obtained from the experimental measurements, and the TOSC value was calculated through Equation 1 below.

TOSC = 100-(∫SA / ∫CA × 100)

∫SA = range integrated from the curve of the sample response

(integrated area from the curve of the sample reaction)

∫CA = integrated range from curve of control response

(integrated area from the curve of the control reaction)

For samples that do not have any oxyradical scavenging capacity, ∫SA / ∫CA = 1 and have a value of TOSC = 0. Conversely, when ∫SA → 0, the TOSC value approaches 100. The value of TOSC is compared with the value in the control, so it is theoretically unaffected by the sensitivity of the instrument, reagents used, or other reaction conditions.

GC equipped with Poropack N column and FID (flame ionizing detector) for the detection of ethylene gas was placed in an oven at 60 ° C, injector 180 ° C and detector 180. It was fixed at 캜 and helium was injected into the column at a rate of 30 ml / min as the mobile phase. For the determination of the ethylene gas in the vial, 150 μl of air in the vial was taken with a gas-tight syringe using the head-space technique, and then gas-tight. The test was performed by injecting a syringe into an injector.

sample Peroxyl Radical Hydroxyl Radical Peroxynitrite (TOSC / mg) Pine Leaf Extract 3227 ± 727 410 ± 18 1076 ± 154 Leek extract 1895 ± 193 429 ± 17 824 ± 182 Glutathione 940 ± 93 257 ± 27 347 ± 3

sample Peroxyl radical Hydroxyl radicals Peroxide TOSC / ml plasma Pine needles (n = 5) 8838 ± 1888 (152) * 11418 ± 504 (106) 6833 ± 1019 (130) *

Antioxidant activity of pine needle extract, leek extract as a negative control and glutathione as a positive control are shown in Table 1 above.

As shown in Table 1, the peroxyl radical scavenging effect of the pine leaf extract showed 1.7-1.9 times the scavenging effect compared to the leek extract. The TOSC value for the peroxyl radical of glutathione, a typical antioxidant in the body, was about 940 TOSC / mg, about 2 times the leek extract, and 3-4 times the pine leaf extract (FIG. 1, FIG. 2 and 3).

When the hydroxyl radical scavenging effect was compared with the TOSC value of glutathione, about 260, the TOSC values of the pine leaf extract and the leek extract showed about 1.6-1.7 times the scavenging effect (see FIGS. 4, 5 and 6).

The scavenging effect of pine needle extract and leek extract did not show significant difference on peroxide. However, it showed about 2.4-3.1 fold activity compared to the TOSC value of glutathione (see FIGS. 7, 8 and 9).

The antioxidant activity analyzed in the plasma obtained from pigs who consumed pine nuts for 4 weeks is shown in Table 2 above.

As shown in Table 2, the results of the experiment, it was confirmed that the TOSC value / ml plasma of the peroxyl radical, hydroxyl radical and peroxide nitrate are 8838, 11418 and 6833 TOSC / ml, respectively.

All results were expressed as mean ± standard deviation, and Student's t-test was performed to verify statistical significance.

As described above, the method of raising animals using the pine needles ( Pinus Koraiensis ) of the present invention and the functional products produced using the same may have an antioxidant effect and contribute to increase of farm income.

Claims (6)

A first step of collecting and grinding the pine leaves; A second step of combining the pine needles and the feed at a composition ratio of 1 to 5: 15 to 20 by weight (w / w); Animal breeding method using the pine nut leaf having an antioxidant activity comprising the third step of feeding the animal feed formulated with the pine leaves to be free vegetarian. delete Pork produced by the animal breeding method using a pine leaf having the antioxidant activity according to claim 1. delete delete delete

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