Feed additive for ruminants

The present invention has an object to provide an economically advantageous rumen bypass formulation capable of containing a biologically-active substance at a high concentration by means of using a protective material composted of safe natural product to ecosystem. The preparation of the rumen bypass formulation for ruminants according to the present invention can be accomplished by dispersing a biologically-active substance in an amount of from 50 to 90% by weight relative to the weight of the formulation into a protective material comprising specific salts of aliphatic carboxylic acid and either of fatty acid or aliphatic alcohol at a ratio of the former to the later being in a range of from 30:70 to 10:90.

This application is a 371 of PCT/JP96/01627 filed Jun. 14,1996. 
1. Field of the Invention 
The present invention relates to a feed additive and, more particularly to 
a matrix-type rumen bypass formulation of feed additives wherein a 
biologically-active substance nutritiously useful for ruminants is 
dispersed in a protective material for protecting said biologically-active 
substance. 
2. Background Art 
A rumen bypass formulation for ruminants is defined as a formulation of 
feed additives which comprises one or more of amino acids, vitamins and 
other biologically-active substance and has a specific characteristic to 
allow elution and absorption of said biologically-active substance in 
digestive organs of ruminants from their fourth stomach onward while 
restricting elution and microbial decomposition of said substance in their 
rumen. 
In breeding of ruminants, it is well accepted from dietetic and clinical 
point of view to feed them together with biologically-active substance 
contained in a rumen bypass formulation. In the practices, the 
incorporation of a biologically-active substance at high concentration to 
a rumen bypass formulation is advantageous and favorable, economically. 
Where, the hardness of the formulation is very important in view of 
durability to a mixing process with feeds and to chewing of cows. However, 
when increasing the concentration of biologically-active substance in the 
formulation, it is required to reduce the content of a protective material 
in the formulation, which has therefore made difficult to maintain rumen 
bypass property and hardness of the formulation. In order to solve this 
problem, a protective material having higher performance than the previous 
be inevitably required. Under the absence of such protective material, the 
concentration of a biologically-active substance in a matrix-type rumen 
bypass formulation has been obliged to be less than 50%. 
The concept and many practical examples of the rumen bypass formulation has 
been already publicly known, however, there is no example for a 
matrix-type formulation which can contain a biologically-active substance 
at a high concentration of more than 50% (high concentrate formulation in 
the market). An example which uses salts of aliphatic monocarboxylic acids 
(fatty acids) as a protective material has been disclosed, however, no 
example which contains a biologically-active substance at a high 
concentration of more than 50% and can give good rumen bypass property, 
has not been disclosed. 
In Japanese Patent Laid-opened No. Hei 2-163043, a concept for a 
matrix-type formulation which uses fatty acid salts and a fatty compound 
compatible to the salt as a protective material is disclosed, however, the 
concentration of a biologically-active substance contained therein is 
lower than 10%, and wherein no concrete description on the formulation 
containing more than 50% of the biologically-active substance, is given. 
On the other hand, a example wherein both fatty acid calcium and stearyl 
alcohol were used as a protective material at a combining rate of 58:2 
(97:3) is disclosed in International Patent Open No. WO/12731, however, 
the concentration of the biologically-active substance is yet lower than 
50%. 
In Japanese Patent Laid-opened No. Sho 56-154956, a matrix-type formulation 
which uses fatty protective material is disclosed, however, the content of 
the biologically-active substance is lower than 50% and the composition of 
the protective material is different from the one given in the present 
invention. 
In U.S. Pat. No. 5,425,963, high purity fatty acid salt as a feed additive 
and the manufacturing method are disclosed, however, the object of that 
invention is different from that of the present invention and the content 
of the biologically-active substance is also far different from the one of 
the present invention. 
Considering such difficulty existing in the prior arts, the present 
invention has an object to provide an economically advantageous rumen 
bypass formulation which can contain a biologically-active substance at a 
high concentration and is using safe natural materials to ecosystem, such 
as fats and oil, and wax, as the protective material. 
DISCLOSURE OF THE INVENTION 
The inventors of the present invention has previously disclosed a rumen 
bypass formulation comprising a biologically-active substance and using a 
protective material as the matrix composed of a fatty acid salt and a 
fatty compound compatible to the salt in WO/12731. Following thereto, the 
inventors of the present invention has further found that the composition 
of the protective material which can remain the concentration of a 
biologically-active substance in the formulation at high during its 
passage though rumen is quite different from the one capable of protecting 
the substance contained at a low concentration and is limited to a certain 
range, and that only a protective matrix comprising aliphatic 
monocarboxylic acid salt and either of aliphatic carboxylic acid or 
aliphatic alcohol, which are combined at a specific composition rate, can 
allow to prepare a rumen bypass formulation that can contain a 
biologically-active substance at a high concentration of more than 50%. 
Therefore, the prevent invention is directed to a rumen bypass formulation 
for ruminants wherein biologically-active substances are dispersed at a 
range of from 50 to 90% by weight relative to the weight of the 
formulation in a protective material I! as described below, of which 
content ranging from 10 to 50% by weight relative to the weight of the 
formulation. 
The protective material I! is composed of both compounds 1) and 2), and 
their combination ratio by weight of the compounds 1) to 2) is in a range 
of from 30:70 to 10:90. 
Compound 1): the compounds are at a least one selected from a group 
consisting of a), b) and c) specified below. 
a) Saturated or unsaturated aliphatic monocarboxylic acid containing 8-24 
carbon atoms and being in either straight- or branched-chain. 
b) Saturated or unsaturated aliphatic alcohol containing 8-24 carbon atoms 
and 1 hydroxy group, and being in either straight- or branched-chain. 
c) Saturated or unsaturated di- or tri-carboxylic acid containing 2-8 
carbon atoms in either straight- or branched-chain. 
Compound 2): the salts of saturated or unsaturated aliphatic monocarboxylic 
acid containing 12-24 carbon atoms and being in either straight- or 
branched-chain. 
Provided, a term, "% by weight", specified in the present invention means 
"% by dry weight", which is calculated based on a weight of a compound 
from which an amount of absorbed-water, that is water be eliminated from 
the compound by drying under ordinary temperature condition at a range of 
from 80.degree. to 120.degree. C., is subtracted. 
The present invention is further described in detail in the following. 
The biologically-active substance used in the present invention is defined 
as a substance which gives any biological activity when it is applied to 
ruminants, and is directed to a substance that is hard to be digested and 
absorbed efficiently by ruminants due to its easy decomposition in their 
rumen when it is administrated orally, such as amino acids including 
methionine and lysine hydrochloride, amino acid derivatives including 
2-hydroxy-4-methylmercaptobutyric acid and the salts thereof, vitamins 
including nicotinic acid, nicotinic acid amide, vitamin A and vitamin E, 
saccharides including grape sugar and fruit sugar, various veterinary 
drugs including antibiotics and anthelmintics. 
The biologically-active substance may be used either alone or in a 
combination of 2 or more of them for the rumen bypass formulation. 
The amount in total of the biologically-active substance to be contained is 
in a range of from 50 to 90% by weight and, preferably in a range of from 
60 to 85% by weight. It is less economical if the amount of the substance 
be less than this range, while the rumen bypass property deteriorates and 
the manufacturing of the formulation becomes difficult when the amount of 
the substance be more than this range. 
The protective material used in the present invention substantially 
comprises a fatty acid salt and either of aliphatic (mono-, di- or 
tri-)carboxylic acid or aliphatic alcohol. The amount of the protective 
material to use is subjected to both of the amount of the 
biologically-active substance and that of a reforming agent that may be 
added when appropriate, but an adequate amount could be determined within 
a range of from 10 to 50% by weight. The fatty acid salt is called as 
"bypass fats and oil" as well, which has a property not to be decomposed 
in the rumen but to be digested in a digestive organs from the fourth 
stomach onward and is the main component of the protective material in the 
formulation. The amount of the fatty acid salt to use is in a range of 
from 70 to 90% by weight relative to the weight of the protective 
material. Out of this range, it is not possible to get a rumen bypass 
formulation which can contain a biologically-active substance at a high 
concentration and can assure excellent rumen bypass property. 
The fatty acid salt used in the present invention is a salt of saturated or 
unsaturated aliphatic monocarboxylic acid containing 12-24 carbon atoms 
and being in either straight- or branched-chain. If the number of the 
carbon atoms is less than that range, the rumen bypass property of the 
formulation will be deteriorated, while digestion capability in digestive 
organs from the fourth stomach onward will decline when the number is more 
than that range. For the examples of said aliphatic monocarboxylic acid, 
lauric acid, palmitic acid, myristic acid, stearic acid, oleic acid, 
linoleic acid and linolenic acid are exemplified, and one or more of these 
aliphatic monocarboxylic acids can be used for the formulation. 
Particularly, a mixture of acids which are originated in animals or 
plants, such as fatty acids obtained from palm oil or beef tallow, is 
preferable in view of commercial availability. 
For the examples of the fatty acid salts, calcium salts, magnesium salts, 
aluminium salts and zinc salts of aliphatic monocarboxylic acids 
containing carbon atoms in a range described above are exemplified, 
however, it is preferable to use any one of the calcium salts. 
In the present invention, it is preferable to use a highly purified fatty 
acid salt, of which purity in the solid component (hereinafter abbreviated 
as "the purity") be preferably more than 90%. The purity used here is 
defined as the proportion of insoluble residue obtained after an 
extraction of the fatty acid salt according to a customary analytical 
method for fats and oil with a solvent, such as ethers and ketones, which 
is calculated by subtracting the amount of absorbed water therefrom. In 
this extraction, however, it is necessary to use a solvent which does not 
dissolve the fatty acid salt but can dissolve fats and oil contained in 
the salt. 
It is preferable that the amount of a base, such as calcium, contained in 
the fatty acid salt can remain at a level approximately equivalent or 
excess in term of physical property, such as hardness. In case that the 
base is calcium salt, it is preferable to contain the base at a rate of 
7-12% by weight as a content of calcium, and preferably 8-10% by weight. 
For the quantitative analysis of calcium, a known analytical method can be 
employed, however, the quantity of calcium is normally determined by 
incinerating the fatty acid salt and consequently analysing the amount of 
calcium contained in the ash obtained. 
Fatty acids obtained from beef tallow and palm oil, which are usable as a 
raw material for the fatty acid salt, normally contain triglycerides at a 
rate ranging from 5 to 40%, and other compounds, such as reaction 
controlling agents and stabilizers, are further added thereto sometime. 
These compounds may remain in the reacted-product of fatty acid salt as 
unreacted components and those become the impurities for the fatty acid 
salt. Some of the fatty acid salts commercially available contain more or 
less 20% of impurities. In the highly concentrated formulation, such 
impurities work to deteriorate the rumen bypass property and the hardness 
of the formulation, and therefore, it is preferable to use highly-purified 
fatty acid salts. 
Aliphatic carboxylic acids and aliphatic alcohols used as the protective 
material of the present invention together with said fatty acid salt are 
considered as a component to improve the compatibility between the 
biologically-active substance and the protective matrix as well as a 
component to reduce the degree of crystallization of the fatty acid salt, 
and it is preferable to use these carboxylic acids and alcohols at a rate 
compatible to the fatty acid salt. Concerning the melting point of these 
carboxylic acids and alcohols, there is a tendency that the melting point 
is preferably close to ruminant's body temperature. The amount range to 
incorporate these carboxylic acid or alcohol in the formulation is between 
10 to 30% by weight relative to the weight of the protective material. Out 
of this range, it is difficult to prepare the formulation while assuring 
good rumen bypass property. 
The number of carbon atoms contained in aliphatic monocarboxylic acids used 
in the present invention is in a range of from 8 to 24, and preferably 
from 12 to 18. If using the acid containing less carbons, the formulation 
becomes soft and it deteriorates the rumen bypass property, while the 
digestion capability in digestive organs from the fourth stomach onward 
deteriorate when using the acid containing carbon atoms more than that 
range. 
For the examples of said aliphatic monocarboxylic acids, caprylic acid, 
capric acid, lauric acid, myristic acid, palmitic acid, palmitoleic acid, 
stearic acid, oleic acid, linoleic acid, linolenic acid, behenic acid, 
hydrogen-added fatty acid of castor oil, and the mixture thereof, are 
exemplified. Most of the aliphatic monocarboxylic acids commercially 
available are saponificated and purified oil originated from animals or 
vegetables. 
The number of carbon atoms contained in the monovalent aliphatic alcohol 
having one hydroxy group used in the present invention is preferably in a 
range of from 8 to 24, and more preferably from 12 to 18, since the 
softening of the formulation and a decline in rumen bypass property are 
recognized when the alcohols having less carbon atoms than that range, 
while deterioration in degree of digestion in digestive organs from the 
fourth stomach onward is observed when the alcohols having carbon atoms 
more than that range. 
For the examples of the aliphatic alcohols used in the present invention, 
octanol, nonanol, dekanol, undekanol, lauryl alcohol, myristic alcohol, 
cetyl alcohol, stearyl alcohol, eicosanol, docosanol, dodecenol, fiseteryl 
alcohol, zoomaryl alcohol, oleyl alcohol, gadoleyl alcohol and the isomers 
thereof, are exemplified. 
The number of carbon atoms of the aliphatic di- or tri-carboxylic acids 
(having 2 to 3 carboxyl groups) used in the present invention is 
preferably in a range of from 2 to 8, and more preferably from 2 to 6, and 
it is difficult to prepare the formulation having good physical property 
with the acid containing carbon atoms out of that range. 
For the examples of the said aliphatic di- or tri-carboxylic acids, oxalic 
acid, malonic acid, succinic acid, glutaric acid, adipic acid, pimelic 
acid, suberic acid, malic acid, citric acid, and the like are exemplified. 
In the present invention, wax, such as rice wax, carnauba wax and beeswax, 
ethyl cellulose, propyl cellulose, polyethylene, chitosan and their 
derivatives, various polymers such as pH-sensitive polymers, powder of 
organic and inorganic materials, and various additives, such as 
stabilizers and perfumes, can be added to the formulation as an improving 
agent for any of the formulation-forming property, the mechanical 
intensity and other properties when appropriate. In addition, it is also 
possible to improve the property of the formulation by coating it with 
such improving agents. 
For the manufacturing of the rumen bypass formulation specified in the 
present invention, various known methods for granulation can be employed. 
However, it is preferable to employ extruding granulation method, and 
wherein it is further preferable to respectively subject the formulation 
to quenching treatment immediately after both processes of evacuation and 
granulation by using either water or cool wind in order to obtain a 
formulation of less voids and less water content. 
Although there is no limitation in the shape of the formulated product to 
manufacture, it is preferable to make it into granules with less corners, 
namely, any of globular, elliptic, cannonball-shaped, and cylindrical 
shapes. Regarding the size of the formulated product, any size appropriate 
for the use as feeds can be selected, however, it is preferable to select 
a size in a range of 0.5 to 10 mm in both diameter and length, which are 
classified into a standard category of granules or pellets.

BEST MODE FOR CARRYING OUT THE INVENTION 
The present invention is further described in detail with referring to the 
following embodiments and examples for comparison. 
However, the scope of the present invention should not be limited to the 
description given in the following embodiments. 
EXAMPLE 1 
28 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 7 parts by weight of palmitic acid and 65 parts by weight of 
methionine were mixed. The mixture was placed in a hopper of a 
double-shaft extruding granulator and was extruded in fused state through 
a die with a diameter of 2 mm while subjecting it to evacuation. 
Then, the mixture extruded was cut with a water-cooled cutter, dehydrated 
by using a centrifuge, and dried by blowing at room temperature to thereby 
obtain a rumen bypass formulation in approximately cylindrical shape 
having an average diameter of 2 mm and an average length of 2 mm. 
EXAMPLE 2 
28 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 5 parts by weight of lauric acid and 70 parts by weight of 
methionine were mixed. The mixture was placed in a hopper of a 
double-shaft extruding granulator and was extruded in fused state through 
a die with a diameter of 1.2 mm while subjecting it to evacuation. Then, 
the mixture extruded was cut with a water-cooled cutter to obtain a 
formulated product in cannonball shape having an average maximum diameter 
of 1.2 mm and an average length of 1.2 mm. After subjecting it to a 
centrifuge, 100 parts by weight of the formulated product and 1 part by 
weight of powder talc were mixed, and the mixture was then placed in an 
oven maintained at 60.degree. C. to dry it for 16 hours, thereby a desired 
rumen bypass formulation was obtained. 
EXAMPLE 3 
23 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 4 parts by weight of lauric acid, 71 parts by weight of methionine 
and 1 part by weight of ethyl cellulose were mixed. The mixture was placed 
in a hopper of a double-shaft extruding granulator and was extruded in 
fused state through a die with a diameter of 1.6 mm while subjecting it to 
evacuation. Then, the mixture extruded was cut with a water-cooled cutter 
to obtain a formulated product in approximately cylindrical shape having 
an average diameter of 1.6 mm and an average length of 1.6 mm. After 
subjecting it to a centrifuge, the formulated product was dried at 
40.degree. C. for 16 hours to thereby obtain a desired rumen bypass 
formulation. 
EXAMPLE 4 
32 parts by weight of calcium salt of palm fatty acid (purity: 97.1%), 5 
parts by weight of myristic acid, 62.5 parts by weight of methionine and 
0.5 part by weight of vitamin E acetate were mixed. The mixture was placed 
in a hopper of a double-shaft extruding granulator and was extruded in 
fused state through a die with a diameter of 1.2 mm while subjecting it to 
evacuation. Then, the mixture extruded was cut with a water-cooled cutter 
to obtain a formulated product in approximately cylindrical shape having 
an average diameter of 1.2 mm and an average length of 1.2 mm. To the 
formulated product, 1 part by weight of bone meal was then added, and the 
mixture was slowly passed for 40 min. through a rotating kirn maintained 
at 50.degree. C. The mixture was then cooled by blowing to room 
temperature, thereby a desired rumen bypass formulation was obtained. 
EXAMPLE 5 
20 parts by weight of calcium salt of palm fatty acid (parity: 94.0%), 4 
parts by weight of lauric acid, 1 part by weight of glyceryl monostearate, 
65 parts by weight of methionine and 10 parts by weight of lysine 
hydrochloride were mixed. The mixture was placed in a hopper of a 
double-shaft extruding granulator and was extruded in fused state through 
a die with a diameter of 1.2 mm while subjecting it to evacuation. Then, 
the mixture extruded was cut with a water-cooled cutter to obtain a 
formulated product in approximately cylindrical shape having an average 
diameter of 1.2 mm and an average length of 1.2 mm. The formulated product 
was then slowly passed for 50 min. through a rotating kirn maintained at 
50.degree. C., and was cooled by blowing to room temperature, to thereby 
obtain a desired rumen bypass formulation. 
EXAMPLE 6 
22 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 6 parts by weight of stearyl alcohol, 1 part by weight of carnauba 
wax and 73 parts by weight of methionine were mixed. The mixture was 
placed in a hopper of a double-shaft extruding granulator and was extruded 
in fused state through a die with a diameter of 1.2 mm while subjecting it 
to evacuation. Then, the mixture extruded was cut with a water-cooled 
cutter to obtain a formulated product in approximately cylindrical shape 
having an average diameter of 1.2 mm and an average length of 1.2 mm. 
After subjecting it to a centrifuge, the formulated product was dried in 
an oven maintained at 40.degree. C. for 16 hours to thereby obtain a 
desired rumen bypass formulation. 
EXAMPLE 7 
29 parts by weight of calcium salt of palm fatty acid (purity: 97.1%), 6 
parts by weight of cetyl alcohol and 65 parts by weight of methionine were 
mixed. The mixture was placed in a hopper of a double-shaft extruding 
granulator and was extruded in fused state through a die with a diameter 
of 2 mm while subjecting it to evacuation. Then, the mixture extruded was 
cut with a water-cooled cutter, dehydrated by using a centrifuge, and 
dried by blowing at room temperature to thereby obtain a desired rumen 
bypass formulation in cannonball shape having an average diameter of 2 mm 
and an average length of 2 mm. 
EXAMPLE 8 
29 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 3 parts by weight of lauryl acid, 3 parts by weight of palmitic 
acid and 65 parts by weight of methionine were mixed. The mixture was 
placed in a hopper of a double-shaft extruding granulator and was extruded 
in fused state through a die with a diameter of 2 mm while subjecting it 
to evacuation. Then, the mixture extruded was cut with a water-cooled 
cutter, dehydrated by using a centrifuge, and dried by blowing at room 
temperature to thereby obtain a desired rumen bypass formulation in 
cannonball shape having an average diameter of 2 mm and an average length 
of 2 mm. 
EXAMPLE 9 
29 parts by weight of calcium salt of palm fatty acid (purity: 97.1%), 4 
parts by weight of myristic acid, 2 parts by weight of succinic acid and 
65 parts by weight of methionine were mixed. The mixture was placed in a 
hopper of a double-shaft extruding granulator and was extruded in fused 
state through a die with a diameter of 2 mm while subjecting it to 
evacuation. Then, the mixture extruded was cut with a water-cooled cutter, 
dehydrated by using a centrifuge, and dried by blowing at room temperature 
to thereby obtain a desired rumen bypass formulation in cannonball shape 
having an average diameter of 2 mm and an average length of 2 mm. 
EXAMPLE FOR COMISON 1 
35 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%) and 65 parts by weight of methionine were mixed. The mixture was 
placed in a hopper of a double-shaft extruding granulator and was extruded 
in fused state through a die with a diameter of 2 mm while subjecting it 
to evacuation. Then, the mixture extruded was cut with a water-cooled 
cutter, dehydrated by using a centrifuge, and dried by blowing at room 
temperature to thereby obtain a desired rumen bypass formulation in 
approximately cylindrical shape having an average diameter of 2 mm and an 
average length of 2 mm. 
EXAMPLE FOR COMISON 2 
20 parts by weight of calcium salt of beef tallow fatty acid (purity: 
97.3%), 15 parts by weight of stearic acid and 65 parts by weight of 
methionine were mixed. The mixture was placed in a hopper of a 
double-shaft extruding granulator and was extruded in fused state through 
a die with a diameter of 2 mm while subjecting it to evacuation. Then, the 
mixture extruded was cut with a water-cooled cutter, dehydrated by using a 
centrifuge, and dried by blowing at room temperature to thereby obtain a 
desired rumen bypass formulation in approximately cylindrical shape having 
an average diameter of 2 mm and an average length of 2 mm. 
EXAMPLE FOR COMISON 3 
20 parts by weight of calcium salt of palm fatty acid (purity: 94.0%), 5 
parts by weight of glyceryl monostearate, 65 parts by weight of methionine 
and 10 parts by weight of lysine hydrochloride were mixed. The mixture was 
placed in a hopper of a double-shaft extruding granulator and was extruded 
in fused state through a die with a diameter of 1.2 mm while subjecting it 
to evacuation. Then, the mixture extruded was cut with a water-cooled 
cutter, dehydrated by using a centrifuge, and dried by blowing at room 
temperature to thereby obtain a desired rumen bypass formulation in 
approximately cylindrical shape having an average diameter of 1.2 mm and 
an average length of 1.2 mm. 
EXAMPLE FOR COMISON 4 
28 parts by weight of calcium salt of palm fatty acid (purity: 94.0%), 2 
parts by weight of stearyl alcohol and 70 parts by weight of methionine 
were mixed. The mixture was placed in a hopper of a double-shaft extruding 
granulator and was extruded in fused state through a die with a diameter 
of 1.2 mm while subjecting it to evacuation. There, the mixture extruded 
was cut with a water-cooled cutter, dehydrated by using a centrifuge, and 
dried by blowing at room temperature to thereby obtain a desired rumen 
bypass formulation in cannonball shape having an average diameter of 1.2 
mm and an average length of 1.2 mm. 
Evaluation on the Formulation Purity of Fatty Acid Salt 
The formulation was subjected to reflux extraction with acetone for 8 hours 
by using a Soxhlet extractor. The purity of fatty acid salt in the 
formulation was calculated from the difference in the dry weight measured 
before and after the reflux extraction. 
Concentration of Calcium contained in Fatty Acid Salt 
After incinerating 1 g of fatty acid salt at 550.degree. C., the ash 
obtained was dissolved in hydrochloric acid, diluted and quantitatively 
analyzed by using emission spectrochemical analyzer (ICP) to determine 
calcium concentration in the fatty acid salt relative to the dry weight of 
said fatty acid salt. 
Hardness 
The hardness of the formulation was measured by using a measurer for the 
hardness of tablets. The hardness was expressed with a value of loading 
pressure at which the formulation starts to destroy. 
The performance of the formulation was also evaluated based on its 
solubility in imitative solutions as described below maintained at 
40.degree. C. by dipping the formulation into each of the solutions in 
turn. 
Eluting Ratio in the First Stomach 
This ratio is to evaluate eluting property of the formulation in the first 
stomach of cows, and which is a ratio of the amount of a 
biologically-active substance, that eluted into the imitative gastric 
juice of the first stomach when the formulation was dipped while shaking 
for 16hours in the said gastric juice prepared at a pH value of 6.4, 
relative to the amount of the biologically-active substance originally 
contained in the formulation. 
Eluting Ratio in the Fourth Stomach 
After measured the eluting ratio in the first stomach, the solid product 
obtained was separated and consequently dipped into imitative gastric 
juice of the fourth stomach of cows prepared at a pH value of 2.0. After 2 
hours shaking, the ratio of the amount of the biologically-active 
substance eluted into the gastric juice relative to the amount of the 
biologically-active substance originally contained in the formulation was 
determined to evaluate the elution property of the formulation in the 
fourth stomach. 
Eluting Ratio in Gastric Juice of Small Intestine 
After measured the eluting ratio in the fourth stomach, the solid product 
obtained was separated and consequently dipped into imitative gastric 
juice of small intestine of cows prepared at a pH value of 8.2. 
After 4 hours shaking, the ratio of the amount of the biologically-active 
substance eluted into the gastric juice relative to the amount of the 
biologically-active substance originally contained in the formulation was 
determined to evaluate the elution property of the formulation in small 
intestine. 
Imitative Gastric Juice of the First Stomach 
This imitative solution is to substitute gastric juice of the first stomach 
of cows, which is prepared by dissolving 2.5 g of disodium 
hydrogenphosphate and 6.7 g of dipotassium hydrogenphosphate into water 
and then adjusting the volume with water to a final volume of 1 l, and of 
which pH value is 6.4. 
Imitative Gastric Juice of the Fourth Stomach 
This imitative solution is to substitute gastric juice of the fourth 
stomach of cows, which is prepared by adding 50 ml of 3.2-N potassium 
chloride and 10 ml of 0.2-N hydrochloric acid to water and then adjusting 
the volume with water to a final volume of 200 ml, and of which pH value 
is 2.0. 
Imitative Gastric Juice of Small Intestine 
This imitative solution was prepared by dissolving 9.8 g of sodium 
hydrogencarbonate, 0.57 g of potassium chloride, 9.30 g of disodium 
phosphate 12H.sub.2 O, 0.47 g of sodium chloride, 0.12 g of sodium sulfate 
heptahydrate, 0.05 g of cow bile powder and 0.05 g of lipase in water and 
consequently adjusting the volume with water to a final volume of 1 l, and 
of which pH value is 
TABLE 1 
__________________________________________________________________________ 
Results in Evaluation of Eluting Ratio 
Example No. 
Example 1 
Example 2 
Example 3 
Example 4 
Example 5 
Example 6 
Example 7 
Example 
Example 
__________________________________________________________________________ 
9 
Details 
of Formulation 
Shape Cylindrical 
Cannonball 
Cylindrical Cannonball 
Outer Diameter- 
2.0--2.0 
1.2--1.2 
1.6--1.6 
1.2--1.2 
1.2--1.2 
1.2--1.2 
2.0--2.0 
2.0--2.0 
2.0--2.0 
Length (mm) 
Biologically- 
Active Substance 
Substance 
Methionine 
Methionine 
Methionine 
Methionine/ 
Methionine/ 
Methionine 
Methionine 
Methionine 
Methionine 
Vitamin E 
Lysine salt 
Conc. (5) 
65 70 71 62.5/0.5 
65/10 73 65 65 65 
Hardness 330 350 290 340 330 300 350 370 340 
Protective Material 
Aliphatic carboxylic 
Palmitic acid 
Lauric acid 
Palmitic acid 
Myristic acid 
Lauric acid Lauric 
Myristic acid 
acid (1) Palmitic 
Succinic acid 
Aliphatic alcohol Stearyl 
Cetyl 
alcohol 
alcohol 
Fatty Acid 
Ca salt (2) 
Raw Material 
Beef suet 
Palm oil 
Beef suet 
Palm oil 
Palm oil 
Beef suet 
Palm oil 
Beef 
Palm oil 
Fatty Acid 
Concentration of Ca 
8.3 7.4 8.3 7.9 7.4 8.3 7.9 8.3 7.9 
Purity (%) 
97.3 94.0 97.3 97.1 94.0 97.3 97.1 97.3 97.1 
Ratio (1):(2) 
20:80 17:83 15:85 14:86 17:83 21:79 17:83 17:83 17:83 
Eluting Ratio 
a) In First 
8 9 10 5 12 14 13 10 8 
Stomach (%) 16 H 
b) In Fourth 
11 23 10 55 43 30 31 15 21 
Stomach (%) 2 H 
c) In Small 
66 58 60 28 31 45 43 58 57 
Intestine (%) 4 H 
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TABLE 2 
__________________________________________________________________________ 
Results in Evaluation of Eluting Ratio 
(Examples for Comparison) 
Example No. 
Example for 
Example for 
Example for 
Example for 
Comparison 1 
Comparison 2 
Comparison 3 
Comparison 4 
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Details of Formulation 
Shape Cylindrical Cannonball 
Outer Diameter-Length (mm) 
2.0--2.0 
2.0--2.0 
1.2--1.2 
1.2--1.2 
Biologically-Active Substance 
Substance Methionine 
Methionine 
Methionine/ 
Methionine 
Lysine salt 
Conc. (5) 65 65 65/10 70 
Hardness 300 280 270 300 
Protective Material 
Aliphatic carboxylic acid (1) 
Stearic acid 
Aliphatic alcohol Stearyl alcohol 
Fatty Acid Ca salt (2) 
Raw Material Fatty Acid 
Beef suet 
Beef suet 
Palm oil 
Palm oil 
Concentration of Ca 
8.3 8.3 7.4 7.4 
Purity (%) 97.3 97.3 94.0 94.0 
Ratio (1):(2) 
0:100 43:57 0:100 7:93 
Eluting Ratio 
a) In First Stomach (%) 16 H 
86 97 57 71 
b) In Fourth Stomach (%) 2 H 
2 1 15 5 
c) In Small Intestine (%) 4 H 
10 1 17 9 
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Application for Industrial Use 
As can be seen from the results shown in Table 1, the eluting ratio of the 
rumen bypass formulation each prepared in the examples from 1 to 9 was low 
in the imitative gastric juice of the first stomach, respectively, while 
the sum of the eluting ratio of the said formulation in both gastric juice 
of the fourth stomach and small intestine was high, respectively. The 
results show an excellent bypass property of these formulations in the 
first stomach and that the formulations can be easily digested in 
digestive organs from the fourth stomach onward. 
Whereas, in Table 2, the formulation prepared according to the referential 
example 1, for which only aliphatic monocarboxylic acid was used as a 
protective material, and all formulations prepared according to the 
Examples for Comparison 2, 3 and 4, for which a protective material out of 
the scope of the present invention was used, respectively, showed high 
eluting ratio in gastric juice of the first stomach and are inferior in 
rumen bypass property, and it is demonstrated that those are less useful 
practically. 
As such, by using protective material comprising a fatty acid salt and 
either of an aliphatic carboxylic acid or a monovalent aliphatic alcohol 
at certain specific composition ratio, a matrix-type rumen bypass 
formulation containing a biologically-active substance at a high 
concentration of more than 50% and having excellent rumen bypass property, 
can be prepared.