KR100305747B1 - Preparation Method for Conjugated Linoleic Acids using Vegetable oils - Google Patents

Abstract

PURPOSE: A process of preparing pure conjugated linoleic acid(CLC) by reacting vegetable oil rich in linoleic acid or linoleic acid in fatty acid state as a substrate under atmospheric pressure or under pressure is provided. By feeding of the CLC composition to livestock in a fatty acid state, animal products having a high content of functional CLC is produced. CONSTITUTION: Fat having a linoleic acid content of 40% or more is dissolved in ethylene glycol and reacted with alkali such as caustic soda or soda ash at 150 to 190deg.C for 30 min to 2.5 hr at 5 to 250psi in 1,000 to 500ml/min nitrogen gas and 30 to 15ml/min hydrogen gas. The reactant is separated by adjusting the pH of the solution to 4.5 to 5.0 and then esterificated. A solution of sulfuric anhydride and ethanol is added to conjugated linoleic acid and then reacted. Oxides contained in the solution are extracted by use of supercritical carbon dioxide.

Description

Preparation Method for Conjugated Linoleic Acids using Vegetable Oils

The present invention relates to a method for producing a conjugated linoleic acid (hereinafter referred to as CLA) composition by isomerization of a vegetable oil-rich raw material rich in linoleic acid. Linoleic acid (LA) is a fatty acid with 18 carbon atoms with double bonds in cis- 9 and cis- 12 configurations, and CLA is a conjugated double bond in cis or trans configuration. It is a generic name for the positional and geometric isomers of linoleic acid with conjugated double bonds. These CLAs also have cis or trans arrays at positions 9, 11 and 10, 12. In theory, these CLAs are eight 9,11 octadecadies by isomerization of linoleic acid ( cis -9, cis -12 octadecadienoic acid). Octadecadienoic acid and 10, 12 octadecadienoic acid ( cis -9, cis -11; cis -9, trans -11; trans -9, cis -11; trans -9, trans -11; cis- 10, cis- 12; cis- 10, trans- 12; trans- 10, cis- 12 and trans- 10, trans- 12) can be formed. However, CLA, which is naturally present in various foods, especially milk and meat produced by ruminant animals, is cis -9 and trans -11 octadicadienoic acid, accounting for 85-95% of the total CLA. (in “Trans Fatty Acids in Human Nutrition” edited by J. L Sebedio and WW Christie. 1998. Chapter 9. 261-302. The Oily press Ltd). However, livestock products produced by unit animals, such as eggs, chicken, pork, quail eggs, dog meat, and duck meat, often have little CLA content. There is only a very small amount of 10 or less. The reason for this is that these functions by the ruminant for animal CLA is an unsaturated fatty acid LA is the Rumen microorganisms of butyric Lee V. blood debris Solvent (Butyrivibrio fibrisolvens) when the unit animal there is a formed of a metabolite of the process in which the hydrogenation reaction This is because there is no microorganism and digestive organs in charge.

On the other hand, US Patent No. 5,770,247 is a method of producing milk with a high natural CLA content by adding 1-5% fat and oil with high content of LA and Linolenic acid. The CLA has several beneficial physiological functions. ① Antioxidant effect is more effective than α-tocopherol and is similar to the effect of BHT. ② As a microbial growth inhibitory effect, Listeria monocyto It is known to be effective against zen ( Listeria monocytogenes) and ③ excellent anti-cancer effect in stomach and breast cancer.

Among these CLAs , cis- 9, trans- 11 and trans- 10, and cis- 12 isomers have strong physiological activity, cis- 9, trans- 11 CLA and trans- 10, cis- 12. CLA also acts as a potent inhibitor of eicosanoid synthesis by inhibiting Δ-9 desaturase activity and reducing PGE ₂ biosynthesis due to structural similarity, thus cis -9, trans -11 and trans The mutual ratio of -10 and cis -12 is known to be important for various physiological effects.

Meanwhile, the related art related to CLA is US Patent 5,070,104; 5,208,356; 5,428,072; 5,430,066; 5,504,114; 5,554,646; 5,585,400; 5,674,901; 5,760,082; 5,760,083; 5,770,247; 5,814,663; 5,827,885; 5,837,733; 5,851,572; 5,856,149; WO 94/16690 (PCT / US93 / 11093) and European Patent 0,440,325 A2. Also, Nichols, PL Jr, J.AOCS, 1951, 73: 247-252, Schneider, WJ, J.AOCS, 1964, 41: 605-606, Hollow, RD, J. AOCS 1963 40: 505 to 506) and gin (Chin, SF, J. Food composition & Analysis, 1992, 5: 185 to 197), etc., are rich in LA, sunflower oil, safflower oil, and LA. When used, alkali isomerization reaction at high temperature can produce several kinds of CLA. CLA production method of US Pat. No. 5,814,663, 5,554,646, etc. is made with 1000g of ethylene glycol, KOH500g, LA or 1,000g of vegetable oil as a substrate, and then acidified to pH 3.0 after 2.5 hours of reaction at 180 ° C., followed by several times of nucleic acid extraction and water or 5% NaCl. After washing with Na ₂ SO ₄ to remove the water and the nucleic acid is evaporated to obtain the CLA. Therefore, when producing CLA in the same manner as described above ① Using an excessive amount of alkali (KOH, NaOH, etc.) of 20% of the total content of ethylene glycol, raw material vegetable oil or LA and alkali as reaction composition ② Many times Extraction and washing process are required. ③ Finally, the produced CLA retains the odor of the used organic solvent, so an additional odor removal process is required. ④ Also, the composition of CLA produced by known methods is cis -9, trans -11 and trans. -10, cis- 12 octadicadienoic acid components are mainly produced, but cis- 9, cis- 11, which are unable to produce CLA with controlled reciprocal ratios of these CLAs, also have no physiological efficacy in CLA; Cis -10, cis -12 and trnas -9, trans -11 octadicanoate were also produced and found to produce 5.9 to 10% of the CLA composition produced. It has been found that the amount of 15-20% of the weight of the final produced CLA is produced together. In particular, these fatty oxides are known to act as a factor for inhibiting animal growth and cancer. U.S. Patent 5,070,104 European Patent 0,440,325 A2 reported that LA and whey protein were used to produce CLA at 85 ° C., but Wolf (Wolff, RL, J. AOCS 1993, 70: 425-430; J. AOCS 1996, 73: 327-332); According to O'Keefe, SF, J. AOCS 1993, 70: 915, the isomerization reaction for the cis-translation of fatty acids occurs only at a temperature of at least 200 ° C, making it difficult to produce the desired CLA quickly. There are disadvantages. In addition, US Pat. No. 5,856,149 can specifically produce cis- 9, trans- 11 CLA using anaerobic butyribibrio fibrisolbene and lactobacilli strains. In order to produce CLA using isomerase produced by microorganisms, LA in free fatty acid form should be used, and culture time is anaerobic, so it is difficult for anyone to apply, and free fatty acid type LA is converted to CLA. Considering the basic medium, such as 36 hours long, there is a disadvantage in the mass production of active CLA, but there is an advantage when used in the fermented milk product using Lactobacillus bacteria. In addition, Korean Patent Publication No. 98-26382 discloses the use of conjugated linoleic acid as feed for chickens and flounder, but the CLA has a high fat oxide content and found that there is a problem that the growth rate is lowered when fed to livestock. To complete.

In order to solve the problems described above, the present invention comprises 98% or more of CLA as a reaction raw material, consisting of an alkali concentration relative to the content of the basic reaction composition, separation of CLA by pH adjustment, an optimum pressure reaction condition, and an oxide removal method. ( cis -9, trans -11 and trans -10, cis -12), the pure CLA product with 100% oxide removal and the ratio of cis -9, trans -11 and trans -10, cis -12 CLA To control and produce CLA quickly and efficiently.

The present invention provides a method for producing a livestock food raw material having high anti-cancer functional CLA content by feeding the CLA composition of the present invention to livestock in the form of fatty acids and then synthesizing it in the body to accumulate CLA in a natural triglyceride form.

1 is a manufacturing process diagram of the present invention.

The present invention is rich in linoleic acid vegetable oil (safflower oil LA content 60-80%, sunflower oil LA content 50-75%, soybean oil LA content 50-57%, corn oil LA content 40-60%, cotton oil LA content 40-52 %) Or fatty acid-like linoleic acid (LA content 60-98%) as a substrate to prepare a CLA 100% of the fatty oxides removed by atmospheric pressure or pressure reaction, cis -9, trans -11 and trans in these CLA -10, cis -12 content can be increased and mutual ratio can be adjusted. In other words, when producing CLA under atmospheric pressure, ethylene glycol, alkali ion concentration, N ₂ input, reaction temperature and time are controlled to produce the reactants with the least amount of fatty oxide and high CLA. It is a method of rapidly producing CLA from which water is removed. In addition, during the pressure reaction, the reaction time was shortened to further reduce the produced fat oxide and optimize the CLA production. In addition, it is possible to control the ratio of cis -9, trans -11, trans -10 and cis -12 of CLA produced by changing the hydrogen input compared to nitrogen input and raw material input, and cis- 9 without physiological function. , cis -11; The amount of cis- 10, cis- 12 and trans- 9, trans- 11 octadicadienoic acid was minimized. The CLA thus produced has a turbid yellow or brown color due to the oxides inevitably produced during the high temperature reaction of fat. Therefore, through the removal of these impurities, a colorless and transparent CLA product with 100% oxide removal can be obtained.

The overall manufacturing process of the present invention is as shown in Figure 1, the following examples and test examples are intended to explain the present invention in more detail, they do not limit the technical scope of the present invention.

Example 1 Preparation of CLA in Linoleic Acid of Safflower Oil Under Atmospheric Pressure

150 g to 75 g of ethylene glycol and 3 g to 25 g of NaOH were placed in a round three-necked flask and bubbling N ₂ to dissolve it at 140 ° C. Then, safflower oil was added in the range of 15 g to 75 g and 140 ° C. to 200 ° C. The reaction is carried out in the temperature range for 30 minutes to 3 hours. After the reaction was completed, the pH was adjusted to 4.5 to pH 5.0 using 6N-HCl solution. Thereafter, the reaction was allowed to stand for 15 to 30 minutes to separate the upper layer, thereby obtaining CLA having a free fatty acid state. The optimum reaction conditions are selected through analysis of the fatty acid composition of each CLA and the oxide content (wt%) per weight of the produced CLA.

Table 1. Production rate of CLA and oxide according to NaOH addition amount (unit:%)

division NaOH (g) 3.1 6.3 9.6 13.0 16.7 20.5 24.4 Linoleic acid 19.4 1.8 1.1 0.9 1.1 1.1 1.1 Total-CLA 57 72 71.5 70.7 67 62 45 oxide 1.2 1.5 1.4 1.8 2.4 4.5 6.8

* Ethylene glycol 150g, safflower oil 15g, temperature 180 ℃ 2 hours

Table 2. Production rate of CLA and oxide according to the amount of safflower oil added (unit:%)

division Safflower Oil (g) 15 25 35 45 55 65 75 Linoleic acid 0.5 0.6 1.3 2.5 25.2 40.7 55.8 Total-CLA 73.7 75.7 74.7 70.1 48.9 31.6 21.0 oxide 2.5 2.8 4.5 6.7 7.0 9.4 10.7

* Ethylene glycol 150g, NaOH 11.3g, temperature 180 ℃ 2 hours conditions

Table 3. Production rate of CLA and oxide according to reaction temperature (unit:%)

division Temperature (℃) 140 150 160 170 180 190 200 Linoleic acid 72.0 49.5 26.8 8.4 1.2 0.8 1.0 Total-CLA 6.9 22.7 40.6 68.4 72.8 70.7 58.4 oxide 0.5 0.7 1.8 3.4 5.5 11.5 15.5

* Condition of 150g ethylene glycol, 40g safflower oil, 11.3g NaOH

Table 4. Production rate of CLA and oxide according to reaction time (unit:%)

division Reaction time (Hr) 0.5 1.0 1.5 2.0 2.5 3.0 Linoleic acid 20.0 5.6 1.5 0.9 0.7 0.7 Total-CLA 54.7 66.6 72.2 74.6 62.4 61.5 oxide - 1.0 3.2 5.3 8.5 15.2

* Ethylene glycol 150g, safflower oil 40g, NaOH 11.3g, temperature 180 ℃

Table 5. Production rate of CLA and oxide according to the amount of ethylene glycol added (unit:%)

division Ethylene glycol (g) 150 125 100 75 Linoleic acid 1.1 1.21 1.3 1.4 Total-CLA 73.5 72.7 73.9 73.5 oxide 5.6 5.7 7.5 10.4

* Condition of safflower oil 40 g, NaOH 11.3 g, temperature 180 ° C. for 2 hours

* Total-CLA in Tables 1,2,3,4,5 refers to cis- 9, trans- 11 and trans- 10, cis- 12 octadecadienoic acid.

Example 2 Production of CLA in Sunflower Oil During Pressure Reaction

315 g of ethylene glycol, 25 g of KOH, and 85 g of sunflower oil were put into a reactor, and N ₂ gas was passed through 1,000 ml to 500 ml / min. At this time, the temperature was raised to 180 ° C and the pressure was reacted in the range of 10 psi to 250 psi, and then adjusted to pH 4.5 to 5.0 with HCl solution, and the reaction was centrifuged to recover CLA to analyze fatty acids and to analyze the amount of oxides produced per CLA weight. In Table 6, the reaction conditions for the optimum oxide content are selected.

Table 6. Formation of CLA and Oxide with Pressure and Reaction Time

division Total-CLA,% Oxide,% Pressure (psi) Response time (hr) Response time (hr) 0.5 1.0 1.5 2.0 0.5 1.0 1.5 2.0 10 52.2 57.0 55.7 55.6 - 0.9 2.2 7.5 20 54.5 56.3 56.3 56.3 - 1.2 1.8 6.8 30 53.8 57.7 56.5 57.1 - 1.3 1.9 8.4 40 52.7 57.7 57.9 58.4 - 0.8 2.0 7.2 50 53.8 57.6 58.0 57.9 - 0.9 2.0 7.9 100 54.4 56.2 56.1 59.4 - 1.0 1.4 8.0 150 56.8 57.8 58.1 58.4 - 1.1 1.5 10.1 200 56.0 57.5 59.0 58.5 - 1.0 1.8 7.4 250 58.3 56.3 58.7 57.6 - 0.9 2.0 8.8

Total-CLA refers to cis- 9, trans- 11 and trans- 10, cis- 12 octadecadienoic acid

Example 3 Production of CLA in which the mutual ratio of cis -9, trans -11 and trans- 10, cis -12 CLA in CLA is controlled in LA during pressure reaction

100 g of LA (content 60%), 350 g of ethylene glycol, and 10 g to 30 g of NaOH were placed in a reactor, while N ₂ gas flowed at 1,000 ml to 500 ml / min and hydrogen (H ₂ ) gas at 30 ml to 15 ml / min. After reacting at 180 ° C. for 1 hour, the mixture was adjusted to pH 4.5˜5.0 with 6N-HCl solution and then centrifuged to recover CLA.

Table 7. Production of cis- 9, trans- 11 and trans- 10, cis- 12 CLA according to NaOH content

% NaOH (g) 0 5 10 15 20 25 30 Linoleic acid 63.1 62.0 60.1 32.5 1.7 1.0 0.8 cis -9, trans -11 CLA - 0.9 2.6 14.4 28.6 42.4 52.0 trans -10, cis -12 CLA - - 0.3 16.2 31.0 18.2 9.2 Reciprocal Ratio of cis -9, trans -11CLA and trans -10, cis -12 CLA - - 1: 8.7 1: 1.13 1: 1.08 1: 0.43 1: 018

Example 4 Removal of Generated Fatty Oxides.

The CLA produced in the above example is brown to yellow because it contains fatty oxides produced as by-products of the reaction. In order to remove 100% of the oxides, the oxides were removed using a supercritical carbon dioxide extractor manufactured by the inventors, and the samples contained 1.5%, 5.5%, 10.7%, and 15.5% of oxides by weight of CLA produced. Used. That is, the structure of the supercritical carbon dioxide extracting device manufactured was made by using a device connected to a CO ₂ cylinder in the order of a cooler, a pressure pump, a heating circulator, a liquid extractor, and an adsorption column for separation. In this case, silica gel, charcoal, and florisil were used as the adsorbent, and the fatty acid ethyl ester type was used as the CLA into the extractor. That is, 1 part of 2% anhydrous sulfuric acid-ethanol solution was added to the CLA to reflux the reaction, and extracted CLA ethyl ester with hexane (Hexane) to remove the solvent in a rotary evaporator was used as a sample. Extraction conditions were carried out at different temperature range (40 ~ 80 ℃) and pressure conditions (500psi ~ 8,000psi). As a result, considering the problems of pyrolysis and polymerization reaction of the synthesized CLA according to the increase of the extraction temperature, the extraction was suitable as low as possible, and the low pressure was efficient when the extraction pressure was considered economical. Table 8-10 summarizes the range of temperature and pressure suitable for CLA extraction and purification among the extraction conditions under the above conditions. The temperature is as low as 40 ~ 60 ℃ range and the pressure is 1,000, 2,500, 4,000 psi. The extraction rate and the oxide content of each extraction fraction were investigated. As a result, in the case of 4,000 psi, the oxides of the extracts were mixed, and the oxide content of each extract was 3.9 to 4.8%, whereas in the case of 1,000 psi, the oxide content of each extract was 1.6-4.5%, indicating a slight oxide removal effect. However, the solubility of CLA in CO ₂ tended to decrease compared to 2,500 psi or 4,000 psi. However, overall, the solubility is appropriate and the oxide content of each extract is 1.8-3.3%, and the oxide removal efficiency is 70-90% when the sample of 15.5% oxide is applied in the pressure range of 1,000psi to 4,000psi. Selection was made to the applicable pressure range. Therefore, we compared the adsorption effect of Florisil, activated charcoal and silica gel as adsorbents using the extraction device in the range of 1,000 ~ 4,000psi which is suitable for supercritical fluid extraction and purification of CLA ethyl ester using supercritical fluid. The same result was obtained. As a result, activated charcoal showed the lowest adsorption effect while extracting three kinds of adsorbents with 5, 25 and 40g per 40g of sample, while silica gel showed the best effect.

Table 8. Extraction of CLA Ethyl Esters and Oxide Contents of Extracts Using Supercritical Fluids at 1,000 psi, 40 ° C

Fraction CO ₂ consumption (ℓ) Extraction amount (g) oxide(%) One 200 0.958 1.6 2 300 5.505 1.3 3 400 4.769 1.2 4 600 4.098 2.0 5 850 5.508 1.7 6 1000 6.441 1.7 7 1500 5.453 2.8 8 2000 3.811 4.5 Unextracted fraction 2.074 34.2

Table 9. Extraction of CLA Ethyl Ester and Superoxide Oxide Contents of Supercritical Fluids at 2,500 psi, 50 ° C

Fraction CO ₂ consumption (ℓ) Extraction amount (g) oxide(%) One 50 4.685 1.8 2 50 9.104 2.5 3 50 6.720 2.1 4 50 4.643 2.3 5 100 6.066 2.4 6 200 5.144 3.3 Unextracted fraction 1.711 21.4

Table 10. Extraction of CLA ethyl ester with 4,000 psi, 60 ° C. supercritical fluid and oxide content of extracts

Fraction CO ₂ consumption (ℓ) Extraction amount (g) oxide(%) One 5 10.815 4.8 2 10 8.675 4.3 3 10 6.829 4.4 4 20 5.789 4.7 5 50 4.832 3.9 Unextracted fraction 1.711 3.0

Table 11. Oxidative Adsorption Effect of Florisil during Extraction of CLA Ethyl Ester Using Supercritical Fluids

Fraction CO ₂ Usage (ℓ) Extraction amount according to the amount of adsorbent (g) Oxide amount according to adsorbent amount (%) 5 25 40 5 25 40 One 50 4.121 4.081 2.925 ND ND ND 2 50 10.785 10.282 9.808 ND ND ND 3 50 7.955 7.848 7.610 0.75 ND ND 4 50 5.121 5.242 4.944 1.36 0.70 ND 5 100 4.989 5.545 6.133 2.28 1.16 ND 6 200 3.044 3.419 4.264 3.33 2.26 0.70 Unextracted fraction 0.512 0.500 0.544 35.33 37.66 34.2 Adsorption powder 1.125 1.315 1.651 82.45 76.43 68.8

Table 12. Oxide adsorption effect of activated charcoal during extraction of CLA ethyl ester using supercritical fluid

Fraction CO ₂ Usage (ℓ) Extraction amount according to the amount of adsorbent (g) Oxide amount according to adsorbent amount (%) 5 25 40 5 25 40 One 50 3.551 3.325 3.161 ND ND ND 2 50 11.666 11.282 10.152 ND ND ND 3 50 7.855 7.948 7.610 0.92 ND ND 4 50 5.821 5.242 5.676 1.75 0.68 ND 5 100 4.889 5.587 5.880 2.88 1.05 0.67 6 200 3.224 3.013 3.631 3.58 1.78 0.68 Unextracted fraction 0.510 0.510 0.500 36.33 36.77 36.5 Adsorption powder 1.325 1.613 1.804 70.30 57.47 48.8

Table 13. Oxide adsorption effect of silica gel on the extraction of CLA ethyl ester using supercritical fluid

Fraction CO ₂ Usage (ℓ) Extraction amount according to the amount of adsorbent (g) Oxide amount according to adsorbent amount (%) 5 25 40 5 25 40 One 50 3.687 3.001 2.851 ND ND ND 2 50 10.012 9.887 9.926 ND ND ND 3 50 7.855 7.654 7.810 ND ND ND 4 50 5.021 5.242 5.876 ND ND ND 5 100 5.897 6.015 5.980 0.88 ND ND 6 200 3.685 3.541 3.331 1.28 ND ND Unextracted fraction 0.541 0.550 0.467 35.33 36.8 42.15 Adsorption powder 1.876 2.024 2.317 65.44 58.09 54.00

Promote the growth of animals using the CLA composition obtained as described above, containing CLA

To produce livestock and produce animal fats containing natural CLA

How it works by feeding broilers, young pigs, laying hens and fattening pigs

Evaluated.

Example 5 Preparation of Feed Composition for Single Cattle

The range of unit livestock feed composition ranges from 45 to 70% for grain feed sources, 5 to 15% for barley feed sources, 0.3% for oil and fat feed sources, 5 to 15% for vegetable protein feed sources, and 2 to 10 for animal protein feed sources. % And mineral feed source, vitamins, amino acids feed, antioxidants, enzymes and probiotics, growth promoters, etc. in the range of 3 to 6% so that the total weight ratio is 100%. In other words, in the CLA-containing feed composition according to the present invention, the above-mentioned CLA composition is mixed with the blended feed in a weight ratio of 0.3% or more, wherein the total fat content of the entire blended feed composition is 3.0% or more. Adjust the blended feed rate. In this case, the content of adding the produced CLA composition to the blended feed is changed according to the specification purpose of the livestock. For example, when 0.3% or more of the CLA composition is blended, the total fatty acid composition contained in the total blended feed is adjusted to Satisfying the indicated CLA content and proportions can achieve the desired effect.

Table 14. CLA Contents and Rates in Blended Feed Compositions

CLA composition dosage,% CLA% in final fatty acid composition Final CLA Rate c-9, t-11 t-10, c-12 ① young broiler 0.3% 1.4 1.5 1: 1.07 1.0% 4.6 4.5 1: 0.98 ② little pig 0.5% 2.3 2.3 1: 1 1.0% 4.3 3.9 1: 0.91 ③ Laying hens 2.5% 8.8 8.5 1: 0.97 5.0% 13.5 13.7 1: 1.01 ④ Laying hens 1.0% 4.6 4.4 1: 0.96 3.0% 11.9 11.2 1: 0.94 5.0% 14.5 13.9 1: 0.96 ⑤ Pigs raised 2.5% 13.5 6.8 1: 0.50 5.0% 20.8 10.2 1: 0.49 ⑥ Native dog 5% 25.8 5.4 1: 0.21 10% 35.8 6.6 1: 0.18 ⑦ Meat duck 2.5% 9.0 6.3 1: 0.70 5.0% 14.5 9.9 1: 0.68 ⑧Quarter 2% 7.5 8.9 1: 1.19 4% 11.1 12.8 1: 1.15

Therefore, grains, etc., contain 2 to 5% of fat in the blended feedstock, so the actual amount of cis -9, trans -11 CLA is at least 1.5%, cis -9, trans -11 in the composition of total fatty acids. And trans -10, cis -12 CLA should be adjusted to the feed composition to a total content of more than 3.0%. This is because the dilution effect occurs in the final blended feed composition when the CLA composition is added by the fat content and different fatty acid composition included in the feed source and fat used, so that the total fat must be obtained in order to obtain the effect using the CLA composition. The total content of cis -9 and trans -11 CLA is more than 1.5% and the total content of cis -9, trans -11 and trans -10 and cis -12 CLA is 3% or more. Feed compositions should be made. The CLA obtained in this way was fed together with the unit feed of the livestock to increase the growth promoting effect, egg production rate and egg shell thickness of young broilers and pigs. Egg production with high CLA, production of natural CLA-containing yolk oil with a controlled ratio of cis -9, trans -11 CLA and trans -10, cis -12 CLA, production of pork with high CLA content, production of pork with CLA, domestic breeding Changes in fatty acid composition in meat, fatty acid composition and cholesterol lowering effect when CLA was added to broiler duck feed, and CLA addition to egg diets were observed.

Table 15. Comparison of CLA Contents of the Invention and Conventional Methods

Conventional method Method herein Linoleic acid substrate (LA content 95%) Safflower oil substrate (LA content 75%) Linoleic acid substrate (LA content 95%) Safflower oil substrate (LA content 75%) C18: 2 ω6 1.4 2.5 0.5 1.1 c-9, t-11 43.3 29.1 65.64 35.15 t-10, c-12 45.3 32.5 26.26 35.85 * c-9, t-11 + t-10, c-12 88.6 61.6 91.9 71.0 c-9, c-11 1.9 2.4 0.85 0.95 c-10, c-12 1.4 2.7 0.75 1.00 t-9, t-11 2.6 4.8 0.65 0.95 Oxide content,% 17.28 15.20 2.25 4.95 the ratio of c-9, t-11 and t-10, c-12 1: 1.05 1: 1.12 1: 0.4 1: 1.02

CLA without physiological functionality, ie cis- 9, cis- 11, as shown in Table 15; The production rate of cis -10, cis -12, trans -9, and trans -11 was 5.9% to 9.9% in the conventional method, but it can be reduced by 62% to 71% in the range of 2.25% to 2.9% in the present method. It is also effective to increase the control and content of the mutual production rate of cis -9, trans -11 and trans -10, cis -12 to be produced.

In addition, the CLA mixture produced as a substrate is subjected to an oxide removal process to remove 100% of the oxide, and also has the effect of removing more than 95% of CLA without biological function, and to produce colorless transparent CLA.

<Test Example>

In the present invention, the CLA content contained in each sample before and after the removal of the oxide was used by gas chromatography (Huwlett Packard 6890 Model GC), the length of the column (Supelcowex 10 Fused Silica Capillary Column: Supelco cat No. 2-4082) A capillary column of 60 m, inner diameter 0.32 mm, and film thickness 0.25 m was used. The column was mounted on GC, and then maintained at an initial temperature of 170 ° C. for 0.5 minutes, and raised at 2.5 ° C. per minute to maintain a final temperature of 225 ° C. for 5 minutes. At this time, the oven temperature was 230 ° C, the detector temperature was 250 ° C, and the carrier gas was helium. 45 mg of the sample for analyzing the CLA content was taken into a cap tube, and 15 ml of a solution containing 2 ml of sulfuric acid and ethyl alcohol and 230 ml respectively was taken into a tube and mixed with nitrogen gas. After that, the tube was reacted for 10 minutes in an 80 ° C. water bath to extract CLA derivatives with 3 ml of nucleic acid, and injected 2 µl into GC. Oxygen-free CLA was added to the feed at 0.5-5.0% to the livestock to achieve the desired effect. In other words, young livestock with low resistance to the external environment showed excellent growth promoting effects, and even when fed the CLA composition before the removal of oxide, it was improved than the results of the general feed fed. In addition, when eggs were produced, the productivity of eggs was improved, and the quality of egg shells produced was also greatly improved, and eggs containing 600 mg to 675 mg / 100 g of natural CLA were produced. Pork can produce pork containing 10% of CLA, cis- 9 and trans- 11 CLA. Therefore, the CLA composition is apparent in the intended effect when fed to livestock, it was able to produce meat and eggs rich in functional CLA and very low in fat and cholesterol content when applied to domestic dogs, duck meat, quail feed.

The conjugated linoleic acid from which the fatty oxides of the present invention are removed is a colorless transparent CLA in which more than 95% of the CLA having no physiological activity is removed, and cis- 9, trans- 11 CLA and trans- 10, cis- 11 CLA in the final product CLA. The mutual content ratio of can be adjusted. Adding and feeding this CLA to the feed of unit animals can promote the growth of young livestock, the production of eggs with high levels of natural CLA in eggs, and the egg thickness of eggs. In addition, when fed to pigs, it is possible to produce pork with high CLA content and egg yolk oil and pork paper rich in triglycerides in the form of natural triglycerides in the back fat of eggs and pigs. In addition, feeding livestock such as domestic dogs, broiled ducks and laying quails can provide 15-45% less fat and cholesterol and produce livestock with high functional CLA content.

Claims (7)

Dissolving the fats and oils in ethylene glycol, reacting the dissolved fats and oils by adjusting the pressure, alkali content, temperature, reaction time, nitrogen and hydrogen dosage, and separating the reactants by adjusting the pH in the range of 4.5 to 5.0. Ethyl esterification reaction of the separated reactants, reflux reaction by addition of sulfuric anhydride-ethanol solution to conjugated linoleic acid, and ethyl esterification of conjugated linoleic acid. A method for producing conjugated linoleic acid using edible fats and oils, characterized in that the step of extracting an oxide using a supercritical carbon dioxide as a fatty oxide. The fat or oil according to claim 1, wherein the fat or oil has a linoleic acid content of 40% or more in vegetable oil or a linoleic acid in the form of fatty acids, and in the range of 75 g to 150 g of ethylene glycol as a dissolving agent for 15 to 70 g of fat, alkali is caustic soda or A method for producing conjugated linoleic acid using edible oil and fat, wherein the soda ash is in the range of 3 g to 30 g, the temperature is in the range of 150 ° C. to 190 ° C., and the reaction time is in the range of 30 minutes to 2.5 hours. The edible oil and fat according to claim 1 or 2, wherein the pressure is in the range of 5 psi to 250 psi and nitrogen gas is reacted with 1,000 ml to 500 ml / min and hydrogen gas is passed through 30 ml to 15 ml / min. Method for producing conjugated linoleic acid using. The method of claim 1, wherein the oxide extraction temperature is in the range of 40 ℃ to 60 ℃ and the extraction pressure in the range of 1,000 psi to 4,000 psi sample weight to extract the oxide adsorption column filled with Florisil, activated charcoal, silica gel A method for producing conjugated linoleic acid using edible fats and oils, characterized in that the extract is used in the range of 12.5 g to 100 g per 100 g. The oxidized deconjugated linoleic acid according to claim 4 has a mutual content ratio of cis- 9, trans- 11 octadecadienoic acid and trans- 10, cis- 12 octadecadienoic acid in a ratio of 1: 1.1 to 1. A method for producing conjugated linoleic acid using edible fats and oils, characterized in that it is contained in a: 0.2 range. The conjugated linoleic acid obtained in the same manner as described in claims 1 to 5 is added to the feed for livestock by 0.3% or more, and contained in the feed. A method for producing conjugated linoleic acid using edible oil and fat, characterized in that for preparing a feed composition for livestock, characterized in that the total fat content is 3.0% or more. According to claim 6, cis- 9, trans- 11 octadecadienoic acid content of the total fatty acid composition contained in the unit animal feed composition is at least 1.5%, cis- 9, trans- 11 and trans- 10, cis- 10 octadecadienoic acid Method for producing conjugated linoleic acid using an edible oil and fat, characterized in that the content is contained more than 3.0% and the ratio of 1: 1.1 or less.