WO2007126191A1

WO2007126191A1 - Method for preparing ruminally protected choline

Info

Publication number: WO2007126191A1
Application number: PCT/KR2006/004773
Authority: WO
Inventors: Wang Shik Lee; Hyeon Seop Kim; Hyun June Lee; Kwang Seok Ki; Soo Bong Park; Hyun Soo Kim; Sang Hoon Bong; Jeon Soo Hong; Min Kyo Chung
Original assignee: Nuvo B & T Corp; RURAL DEVELOPMENT ADMINISTRATION NATIONAL LIVESTOCK RESEARCH INSTITUTE
Current assignee: Nuvo B & T Corp; RURAL DEVELOPMENT ADMINISTRATION NATIONAL LIVESTOCK RESEARCH INSTITUTE
Priority date: 2006-05-02
Filing date: 2006-11-14
Publication date: 2007-11-08
Anticipated expiration: 2008-11-02
Also published as: KR100698452B1; CN101431903A; JP2009535056A; JP5159766B2; CN101431903B

Abstract

The present invention relates to a method for preparing double-coated ruminally protected choline, which is not degraded by microorganisms, pH and water in the rumen, and thus can be stably delivered and absorbed in the lower digestive tract. More particularly, the method comprises the steps of: (a) preparing carrier-immobilized choline by immobilizing concentrated liquid choline on a carrier and then removing water from the immobilized choline; (b) preparing a first encapsulated choline by encapsulating the carrier-immobilized choline with a binder/coating material; (c) screening encapsulated choline having a size of 0.5-2.0mm from the first encapsulated choline; and (d) further encapsulating the screened encapsulated choline with any one or a combination of two or more selected from the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40°C.

Description

METHOD FOR PREPARING RUMINALLY PROTECTED CHOLINE

TECHNICAL FIELD

The present invention relates to a method for preparing ruminally protected choline, which is not degraded by microorganisms, pH and water in the rumen, and thus can be stably delivered and absorbed in the lower digestive tract. More particularly, the present invention relates to a method for preparing double-coated ruminally protected choline, comprising the steps of: (a) preparing carrier- immobilized choline by immobilizing concentrated liquid choline on a carrier and then removing water from the immobilized choline; (b) preparing a first encapsulated choline by encapsulating the carrier-immobilized choline with a binder/coating material; (c) screening encapsulated choline having a size of 0.5-2.0 mm from the first encapsulated choline; and (d) further encapsulating the screened encapsulated choline with any one or a combination of two or more selected from the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 °C .

BACKGROUND ART

Choline is known as an essential component in studies on some livestock, but the choline requirements for lactating dairy cows have not been established. Also, most choline is degraded in the rumen of ruminant animals, and thus there is a need to develop a technology of protecting choline from degradation such that choline can be passed through the rumen and absorbed and used in the small intestine.

Lactation response to choline is similar to the response of methionine, which is an essential amino acid, and typical symptoms of choline deficiency include muscular weakness, the occurrence of fatty liver, renal bleeding, and the like.

It was found through studies on the injection of choline that choline is a limiting nutrient in diary cows. Also, in choline deficiency, fatty liver occurred due to the fat mechanism and abnormal secretory mechanism of the liver, and when the injection of choline was increased, the migration of fatty acid was increased, thus leading to a reduction in occurrence of fatty liver.

It has been reported that choline affects blood glucose, plasma cholesterol, serum insulin and serum NEFA (nonesterified fatty acids) to significantly contribute to improvements in milk production, milk composition, weight gain and carcass characteristics, but choline chloride supplemented in feed is mostly degraded in the rumen.

Ruminal microorganisms degrade phosphatidyl choline in feed into choline and phosphodiglycerides and also degrade choline by converting choline to methane via trimethylamine as an intermediate metabolite. As a result, less than 1% of choline in feed is known to reach the small intestine.

Choline is an essential nutrient in livestock animals, but the effect thereof cannot be seen in ruminant animals because it is degraded in the rumen. For this reason, attempts to prevent choline components from being degraded in the rumen have long been made.

For example, an encapsulating choline composition for enhancing weight gain in ruminants was reported (US 5,807,594). However, the application of the choline composition as actual products will incur hidger costs due to various materials used as encapsulating agents for the choline composition. Meanwhile, US 5,496,571 discloses an encapsulating choline composition to be fed to lactating dairy cows, and US 5,190,775 discloses that, when a bioactive substance such as choline is fed through a hydrophobic coating, it can be prevented from being degraded in the rumen. However, said patents do not suggest a specific method for preparing the compositions, and simple coating or encapsulation with the hydrophobic substance according to said patent does not provide sufficient protection of active substances from degradation by ruminal microorganisms in the rumen.

Also, WO 94/15480 discloses feeding an encapsulated choline composition to ruminants to increase milk production. However, in this patent, a hydrophobic coating is also used and a method for preparing the choline composition is not specifically mentioned. Furthermore, WO 96/08168 discloses a ruminant feed additive which is prepared by encapsulating a liquid choline compound in a carrier and coating the compound with fatty acid or fatty acid salt. However, there is a shortcoming in that, when the liquid choline is coated with the fatty acid salt, it will react with carbon dioxide in the air.

In addition, a lipid coating (WO 03/059088A1) and a double coating (US 2005/0019413 Al), which make choline stable in the rumen, are known. However, there is a shortcoming in that methods of forming the coatings are complicated.

As described above, many patents relating to choline compositions for enhancing weight gain and milk production are disclosed, and mostly mention that choline is coated with a specific protective substance such that it is not degraded in the rumen. However, these patents do not specifically disclose the method of preparing the choline compositions.

Also, among ruminally protected choline products, which are commercially available currently, there are products prepared by mixing choline with a protective substance and subjecting the mixture to a cold spray process. In this case, however, high costs of factory sites and high installation costs are required, and very cold air having a temperature of -60 °C is used, thus leading to an increase in production cost.

Furthermore, because the products are prepared using a rapid cooling process, macropores can be formed in the products, and operating conditions for producing the products are complicated. In addition, choline located on the surface of the ruminally protected choline products cannot be protected, thus leading to a reduction in the effect of the products.

Thus, there is an urgent need to develop a technology of preparing a coated choline compound, which is stable against ruminal degradation and can be fed to ruminants.

Accordingly, the present inventors have made extensive efforts to solve the problem that choline is degraded in the rumen of ruminants. As a result, the present inventors have found that ruminally protected choline, prepared by immobilizing choline on a carrier, removing water from the immobilized choline, encapsulating the water-free choline with a hydrophobic substance, and then further encapsulating the encapsulated choline with a substance selected from among extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 °C , is stably delivered and absorbed into the lower digestive tract without being degraded by microorganisms, pH and water in the rumen, thereby completing the present invention.

SUMMARY OF INVENTION It is an object of the present invention to provide a method for preparing double- coated ruminally protected choline, which is not degraded by microorganisms, pH and water in the rumen of ruminants.

To achieve the above object, the present invention provides a method for preparing double-coated ruminally protected choline, the method comprises the steps of: (a) preparing carrier-immobilized choline by immobilizing concentrated liquid choline on a carrier and then removing water from the immobilized choline; (b) preparing a first encapsulated choline by encapsulating the carrier-immobilized choline with a binder/coating material; (c) screening encapsulated choline having a size of 0.5-2.0 mm from the first encapsulated choline; and (d) further encapsulating the screened encapsulated choline with any one or a combination of two or more selected from the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 °C .

The above and other objects, features and embodiments of the present invention will be more clearly understood from the following detailed description and accompanying claims.

DETAILED DESCRIPTION OF THE INVENTION, AND PREFERRED EMBODIMENTS

The present invention relates to a method for preparing double-coated ruminally protected choline, which is not degraded by microorganisms, pH and water in the rumen, and thus can be stably delivered and absorbed in lower digestive tract.

According to the present invention, in order to prepare the double-coated ruminally protected choline, concentrated liquid choline is immobilized on a carrier, and water is then removed therefrom, to prepare carrier-immobilized choline. In this regard, the concentrated liquid choline is preferably one or a mixture of two or more selected from the group consisting of choline chloride, choline dihydrogen citrate, tricholine citrate and choline bitartrate. The carrier is preferably a water- insoluble carrier selected from the group consisting of silica, cellulose, starch and zeolite. More preferably, the carrier is silica.

Then, the carrier-immobilized choline is encapsulated with a binder/coating material to prepare granular or needle-shaped, first encapsulated choline. The encapsulation of the carrier-immobilized choline is performed by mixing and agitating the immobilized choline and the binder/coating material in a mixer such as a ribbon mixer, general agitator, a homo-mixer or a V-type mixer, and then extruding the mixture through an extruder or a plodder. Herein, the choline and the binder/coating material are preferably mixed with each other at a weight ratio of 1:0.1-2.0. Also, a perforated plate in the extruder has preferably a size of 0.5-2.5 mm, and an extrusion barrel in the extruder is preferably maintained at a temperature of 10-60 ^"C .

Furthermore, the binder/coating material is preferably any one or a mixture of two or more selected from the group consisting of monovalent soap, divalent soap, and fatty acid having a melting point of more than 40 ^°C . The monovalent soap is preferably sodium soap or potassium soap, and the divalent soap is preferably selected from the group consisting of beeswax, PEG, surfactants, calcium stearate, zinc stearate and magnesium stearate. The fatty acid having a melting point of more than 40 °C is preferably selected from the group consisting of hydrogenated oil, extremely hydrogenated oil, palmitic acid and stearic acid. More preferred is hydrogenated oil.

Then, from the first encapsulated choline, encapsulated choline having a particle size of 0.5-2.0 mm is screened. The screened encapsulated choline is further encapsulated with any one or a combination of two or more selected form the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 °C , thus preparing double-coated ruminally protected choline.

The content of the double coating layer in the finally prepared, double-coated ruminally protected choline is preferably 15-55% (w/w).

The double-coated ruminally protective choline prepared according to the present invention has advantages in that it does not show formation of micropores which is a problem in the products prepared according to the prior commercial cold spray process, and has a uniform shape.

Also, the products prepared according to the cold spray process has shortcomings in that choline is located on the particle surface, and thus has reduced stability in the rumen, whereas the double-coated ruminally protected choline prepared according to the present invention has advantages in that, because the choline- containing product subjected to the first coating (encapsulation) process is further subjected to the second coating (encapsulation) process, the possibility of choline distributed on the product surface is eliminated, and thus most of the choline contained in the product can be maintained stable. In addition, the present invention does not entail the use of a refrigerant used in the cold spray process, thus reducing production expenses, and requires low costs of factory site and installation, so that it reduces the production cost of the product.

When ruminally non-protected choline is fed to ruminants, most of the nonprotected choline will be degraded in the rumen, making it difficult to expect the effect of feeding choline, resulting in wasting choline. On the other hand, when the inventive ruminally protected choline having increased stability is used in livestock breeding, it can increase the nutrient availability of ruminants, thus leading to an increase in the production of livestock, and show increased efficacy in livestock. Thus, the inventive ruminally protected choline can contribute to the development of feed additives capable of producing functional livestock.

Examples

Hereinafter, the present invention will be described in further detail with reference to examples. It is to be understood, however, that these examples are for illustrative purposes only and are not to be construed to limit the scope of the present invention.

Example 1; Preparation of double-coated ruminally protected choline

Concentrated liquid choline was immobilized on a carrier such as silica, cellulose, starch, zeolite or the like, and then water was removed therefrom, thus preparing carrier-immobilized choline .

Then, a binder/coating material such as beeswax, PEG, hydrogenated oil or the like was added to the carrier-immobilized choline, and then mixed and agitated in a mixer such as a ribbon mixer, a general agitator, a homo-mixer, a V-type mixer or the like. The mixture was passed through an extruder or a plodder, thus preparing first encapsulated (coated) choline. Herein, the temperature of an extrusion barrel in the extruder or the plodder was maintained at 10-60°C .

Among the first encapsulated choline, encapsulated choline having a particle size of 0.5-2.0 mm was screened. To the screened choline, one or a combination of two or more selected from the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 ^°C , for example, hydrogenated vegetable oil, hydrogenated corn oil, hydrogenated cottonseed oil, hydrogenated peanut oil, hydrogenated palm kernel oil, hydrogenated palm oil, hydrogenated palm stearic acid, hydrogenated sunflower oil, hydrogenated canola oil, palmitic acid and stearic acid, was added. The mixture containing the first encapsulated choline was subjected to a second encapsulation process in an encapsulating device, such as a high-speed mixer, a ribbon mixer, roedige mixer, or a fluidized bed processor, thus preparing double-coated ruminally protected choline.

Test Example 1

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and 1 kg of the mixture was extruded through a perforated plate having a hole diameter of 1.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high-speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 2

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 3 1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.5 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high- speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 4

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 2.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 5 1 kg of monovalent soap (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high-speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 ^°C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 6

1 kg of PEG (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high-speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 ^°C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 7

1 kg of Tween 80 (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high-speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 ^"C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 8 1 kg of Span 80 (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high-speed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 ^°C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 9

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 500 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 33.3% based on the total weight of the finally produced, second encapsulated material.

Test Example 10 1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 350 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 25.9% based on the total weight of the finally produced, second encapsulated material.

Test Example 11

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 250 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 20.2% based on the total weight of the finally produced, second encapsulated material.

Test Example 12

1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 150 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 "C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 13.0% based on the total weight of the finally produced, second encapsulated material.

Test Example 13 1 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 5 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 1.2 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a high- speed mixer, to which 50 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C, was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 4.8% based on the total weight of the finally produced, second encapsulated material.

Test Example 14

2 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 4 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 2.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 °C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Test Example 15 3 kg of liquid hydrogenated oil (first encapsulating material; binder/coating material) was mixed with 3 kg of choline, and the mixture was extruded through a perforated plate having a hole diameter of 2.0 mm, to prepare a first encapsulated material. 1 kg of the prepared first encapsulated material was placed in a highspeed mixer, to which 670 g of any one or a mixture of two or more selected from among liquid hydrogenated oils, and fatty acids having a melting point of more than 40 "C , was then added as a second encapsulating material, thus preparing a second encapsulated material. Herein, the amount of the second encapsulating material was 40.1% based on the total weight of the finally produced, second encapsulated material.

Example 2: in vitro loss of ruminally protected choline

The in vitro loss of each of the double-coated cholines prepared in Test Examples 1-15 was measured in the following manner, and the measurement results are shown in Table 1 below.

In order to measure the in vitro stability of each of the double-coated cholines prepared in Test Examples 1-15, each of the prepared ruminally protected cholines was placed in a test tube, to which distilled water was added, to prepare a 5% choline aqueous solution. The aqueous solution was stirred using the JEIO TECH MC-I l multi-stirrer at 40⁰C ± 0.1 ^°C for 2 hours.

The stirred aqueous solution was filtered through Whatman paper NO.3 to collect only a solid. The solid was dried at 45 °C ± 0.1 ^°C for at least 12 hours to completely remove the remaining water.

The dried sample was cooled to room temperature in the Sanpla Dry Keeper, and 300 mg of the cooled sample was completely dissolved in a mixed solvent of chloroform and methanol (1:1 v/v). The content of choline in the solution was measured using a 5% K₂CrO₄ solution as indicator, and a 0.1 N AgNO₃ solution as titrant.

The measurement results are shown in Table 1 below. As shown in Table 1, there was little or no change in the in vitro loss of choline according to the change in hole size of the perforated plate (Test Examples 1-4). With respect to the kind of the first encapsulating material, the case of using monovalent soap or divalent soap PEG as the first encapsulating material showed a reduction in in vitro loss compared to the case of using Tween 80 or Span 80. As a result, it could be observed that the use of liquid hydrogenated oil as the first encapsulating material could lead to a reduction in the in vitro loss of choline compared to the use of monovalent soap, divalent soap PEG, Tween 80 or Span 80 (Test Examples 5-8).

With respect to the amount of second encapsulating material added, it could be seen that an increase in the amount of second encapsulating material added led to a reduction in the in vitro loss of choline (Test Examples 9-13).

Also, with respect to the ratio of the first encapsulating material to choline, it was observed that an increase in the ratio of the first encapsulating material to choline led to a reduction in the in vitro loss of choline (Test Examples 14 and 15).

Table 1

As a result, it could be found that the double-coated ruminally protected choline, prepared by selecting liquid hydrogenated oil as the first encapsulating material, mixing choline with the first encapsulating material at a ratio of 1 :0.5 or 1 :1, subjecting the mixture to the first encapsulating process and subjecting the first encapsulated material to the second encapsulating process using an increased amount of the second encapsulating material, showed minimized in vitro loss.

Example 3; Feeding test of ruminally protected choline

A feeding test in vivo was conducted on the ruminally protected choline prepared in Example 15, which showed low in vitro loss in the test of Example 2 and had suitable size and a high choline content.

To conduct the breeding test of diary cows using the ruminally protected choline, 10 diary cows in middle lactation were selected as test animals and fed a sufficient amount of a total mixed ration of compound feed and bulky feed two times daily at 12-hr intervals. Also, the ruminally protected choline (25% of choline content) was additionally fed to the cows in an amount of 40 g (10 g based on choline) per cow when feeding the total mixed ration. The cows were milked two times daily and the amount of milk produced was measured. A milk sample was taken from the produced milk, and the milk components and somatic cell number of the milk sample were measured using an automatic analyzer (LactoScopeR, MK2, Delta Instruments, The Netherlands).

The measurement results are shown in Table 2 below. As shown in Table 2, in the case where the ruminally protected choline was fed to the cows, the daily milk production of the cows increased by 9.9% from 29.2 kg before feeding to 32.1 kg after feeding, and the amounts of milk fat, milk protein, lactose and total solids also increased by 7.9%, 10.4%, 12.7% and 10.1%, respectively. Also, the number of somatic cells in the sample decreased by 61.9% from 167,000 cells/ml to 103,000 cells/ml due to the feeding of the ruminally protected choline, suggesting that the ruminally protected choline was effective in improving milk quality. In addition, the content of choline in the sample increased by about 48.8% from 44.85 mg/ml to 66.75 mg/ml, suggesting that the ruminally protected choline made it possible to produce choline-rich milk.

Table 2

INDUSTRIAL APPLICABILITY

As described in detail above, the present invention provides the method for preparing double-coated ruminally protected choline. The ruminally protected choline prepared according to the present invention is not degraded by microorganisms, pH and water in the rumen, and thus can be stably delivered and absorbed in the lower digestive tract. Also, when it is used in the breeding of ruminant livestock, it can increase the nutrient availability of the ruminant livestock, thus leading to an increase in the livestock production, and makes it possible to produce functional livestock producing choline-rich milk. Furthermore, the inventive method for preparing the ruminally protected choline can be applied to protect various components from ruminal degradation and can be used for the production of a variety of novel feed additives. Although the present invention has been described in detail with reference to the specific features, it will be apparent to those skilled in the art that this description is only for a preferred embodiment and does not limit the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims

THE CLAIMS What is Claimed is:

1. A method for preparing double-coated ruminally protected choline, the method comprises the steps of:

(a) preparing carrier-immobilized choline by immobilizing concentrated liquid choline on a carrier and then removing water from the immobilized choline;

(b) preparing a first encapsulated choline by encapsulating the carrier- immobilized choline with a binder/coating material; (c) screening encapsulated choline having a size of 0.5-2.0 mm from the first encapsulated choline; and

(d) further encapsulating the screened encapsulated choline with any one or a combination of two or more selected from the group consisting of extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40°C .

2. The method for preparing double-coated ruminally protected choline according to claim 1, wherein the concentrated liquid choline is one or a mixture of two or more selected from the group consisting of choline chloride, choline dihydrogen citrate, tricholine citrate and choline bitartrate.

3. The method for preparing double-coated ruminally protected choline according to claim 1, wherein the carrier is a water-insoluble carrier selected from the group consisting of silica, cellulose, starch and zeolite.

4. The method for preparing double-coated ruminally protected choline according to claim 1 , wherein the binder/coating material is any one or a mixture of two or more selected from the group consisting of monovalent soap, divalent soap, and fatty acid having a melting point of more than 40 °C .

5. The method for preparing double-coated ruminally protected choline according to claim 4, wherein the monovalent soap is sodium soap or potassium soap.

6. The method for preparing double-coated ruminally protected choline according to claim 4, wherein the divalent soap is at least one selected from the group consisting of beeswax, PEG, surfactants, calcium stearate, zinc stearate and magnesium stearate.

7. The method for preparing double-coated ruminally protected choline according to claim 4, wherein the fatty acid having a melting point of more than 40 ^°C is selected from the group consisting of hydrogenated oil, extremely hydrogenated oil, palmitic acid and stearic acid.

8. The method for preparing double-coated ruminally protected choline according to claim 1, wherein said extremely hydrogenated oils and fats, and fatty acids having a melting point of more than 40 °C is selected from the group consisting of hydrogenated vegetable oil, hydrogenated corn oil, hydrogenated cottonseed oil, hydrogenated peanut oil, hydrogenated palm kernel oil, hydrogenated palm oil, hydrogenated palm stearin oil, hydrogenated sunflower oil, hydrogenated canola oil, palmitic acid and stearic acid.

9. The method for preparing double-coated ruminally protected choline according to claim 1, wherein said choline and said binder/coating material are mixed with each other at a weight ratio of 1 :0.1-2.0.

10. The method for preparing double-coated ruminally protected choline according to claim 1, wherein the step (b) comprises: mixing and agitating the immobilized choline in a mixer; and then encapsulating in an extruding machine.

11. The method for preparing double-coated ruminally protected choline according to claim 10, wherein the mixer is selected from the group consisting of a ribbon mixer, general agitator, a homo-mixer and a V-type mixer.

12. The method for preparing double-coated ruminally protected choline according to claim 10, wherein the extruding machine is an extruder or a plodder.

13. The method for preparing double-coated ruminally protected choline according to claim 1 , wherein the first encapsulated choline is granular- or needle-shaped.

14. The method for preparing double-coated ruminally protected choline according to claims 10 or 12, wherein a size of perforated plate in said extruding machine is 0.5-2.5 mm.

15. The method for preparing double-coated ruminally protected choline according to claim 12, wherein the temperature of an extrusion barrel in said extruding machine is maintained at 10-60 ^°C .

16. The method for preparing double-coated ruminally protected choline according to claim 1, wherein the step (d) is performed by a second encapsulating device selected from the group consisting of a high-speed mixer, a ribbon mixer, roedige mixer, and a fluidized bed processor.

17. The method for preparing double-coated ruminally protected choline according to claim 1, wherein the content of the double coating layer is 15-55% (w/w).