CA1037768A - Ruminant feed supplement and process for producing same - Google Patents
Ruminant feed supplement and process for producing sameInfo
- Publication number
- CA1037768A CA1037768A CA216,798A CA216798A CA1037768A CA 1037768 A CA1037768 A CA 1037768A CA 216798 A CA216798 A CA 216798A CA 1037768 A CA1037768 A CA 1037768A
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- urea
- glucose
- mixture
- ureides
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Abstract
ABSTRACT OF THE DISCLOSURE
The starch in barley or a like grain is hydrolyzed with the enzyme glucoamylase to produce glucose.
The glucose is then reacted with urea under drying conditions to produce a high assay mixture of ureides, mainly monoglucosyl ureide. The end product is found to be a good non-protein nitrogen feed supplement for cattle.
The starch in barley or a like grain is hydrolyzed with the enzyme glucoamylase to produce glucose.
The glucose is then reacted with urea under drying conditions to produce a high assay mixture of ureides, mainly monoglucosyl ureide. The end product is found to be a good non-protein nitrogen feed supplement for cattle.
Description
`: :
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BACKGROUND OF THE INVENTION
This invention relates to a process for pro-ducing a non-protein nitrogen (NPN) containing material useful as a feed supplement for cattle and sheep. More ; 5 particularly, it relates to a process for producing urease-resistant glucosyl ureides and to the high assay product of the process.
It is now common to feed cattle and sheep feed supplements. These supplements provide nitrogen as a source for meeting the protein needs of the animals.
The most commonly used supplement is urea. When the urea enters the rumen of the animal, the enzyme urease acts on it to form ammonia. The microorganisms in the rumen then convert - the ammonia into a form of protein which can be utilized by - 15 the animal.
There is a disadvantage in using urea in that ; : , it is converted to ammonia at a relatively rapid rate. If the dosage of urea is too large, the capacity of the micro-organisms to convert the produced ammonia to protein is exceeded and the excess ammonia is either converted to urea and ex-creted by the animal or, if its concentration is high enough, ,, I_ , ~ .
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Recent work has been directed toward develop-ing "slow release" NPN compounds. The objective is to :.-provide compounds which produce ammonia in the rumen at a reduced rate with which the microorganisms can cope.
One family of compounds, glucosyl ureides, is known to have the property of slow release of ammonia in the rumen of cattle or sheep. These compounds have the added advantage of providing the ammonia in association with an energy source (glucose), which promotes greater growth of the microflora in the rumen. A United States patent of interest in this connection issued to McNeff under No. 3677767. This patent teaches reacting molasses (which contains glucose and non-reducing sugars, such as fructose) with urea under ~ 15 acidic conditions to produce a liquid product containing ; glucosyl ureide. The product has been shown to be useful as a feed supplement for ruminant animals.
Other pertinent prior art is exemplified by U.S. patents: 2612497, issued to Meijer; 3023205, issued to Meijer; and 3020273, issued to Steadman.
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SUMMARY OF THE INVENTION
: `
.. ,, i The present invention is eoncerned with a - ! known reaction whereby glucose is eombined with urea under aeidie eonditions to produce glucosyl ureides. However, in aeeordance with the invention the process is carried out in a manner sueh that a relatively high degree of eonversion j of the glucose is aehieved, and the end product contains ¦ a relatively high concentration of ureides.
More particularly, glucose is reacted with urea under aeidie and drying eonditions to produee a produet, :' 1037~
.
preferably solid, containing a relatively hi~h proportion o~ urease - resistant glucosyl ureides.
- When first considering drying the glucose-urea Mixture during reaction to drive off water and shift the equilibrium of the reaction toward producing more ureides, we anticipated that the concentration of acid would increase, leading to dehydration and undesired carmelizing or degradation.
Surprisingly, the pH of the acidic mixture actually increases during the drying process. Carmelizing is not a serious problem.
In retrospect, it appears that ammonia, released from the urea, or protein, which is present if one uses hydrolyzed grain as the starting material, buffers the reaction to permit it to proceed to completion.
Since drying is an integral part of the process, we prefer to use a glucose-containing starting material which -is relatively free of other hexoses, which make drying difficult - and which can result in charring of the final product. We have found that grain starch, such as that occurring in barley, wheat or corn, may be hydrolyzed to provide a preferred starting material containing glucose. If, for example, molasses is used in the process, it will not readily dry to a useful product, due to the presence of a mixture of sugars and the tendency of contained fructose to char. However, we have found ; that some hygroscopic mixtures of sugars can be tolerated in the starting mixture, provided that inert materials, such as grain husks, are also present.
~- The product of the process comprises a solid, con-densation reaction product of (1) glucose-containing hydrolyzate of grain, selected from the group consisting of barley, wheat and corn, and (2) urea, in the presence of acid under drying condition at elevated temperature, said product containing the major propcrtion of said glucose in the form of glucosyl ureides.
.
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The process for producing the desired mixture of glucosyl ureides comprises the steps of: providing an aqueous ~ ;
` mixture of a starting material containing one or more reducing carbohydrates as the major dry matter constituent, the major constituent of said reducing carbohydrates being glucose, and urea; and reacting the mixture under acidic and drying con~
ditions at an elevated temperature sufficient to convert the major portion of the glucose and urea into glucosyl ureides ~
and to produce a solid product~ ;
DESCRIPTION OF`THE DRAWINGS
` In the drawing:
' Figure 1 shows a preferred form of the process outlined in terms of chemical equations for the hydrolysis and condensation reactions; and Figure 2 is a plot showing the effect of tempera-' ture of the reactlon mixture on the hydrolysis reaction.
DESCRIPTION OF T~E PREFERRED E~BODIMENT
The invention will now be described in terms of providing glucose by hydrolyzing starch present in barley.
' 20 While a number of grains, such as wheat, corn and the like can be used, barley is preferred as it is commonly used in Western Canada as cattle feed. ' - - , Equipm_ t' _ d`analy's'is pro`c'edur`es The hydrolysis reaction was carried out in a ' ' :' 25 50 ml reactor provided with stirrer, inlets for introduction ' of reagents and sampling, a mantle, and a temperature controller ~'~
capable of keeping the temperature within'+ .5~C. ~ '' The glucose analyses were performed by the ; orthotoluidine method and by the use of a Beckman* ~lucose autoanalyzer, Model ERA-2001*. The first method involves the reaction of the glucose present with o-toluidine in acetic acid.
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The color is measured spectrophotometrically using 5 determin-ations for each sample, one for the blank, and one for the standard. The error of the method was found to be 2%. Time ;
employed for each deter~in~tion: ~15 minutes.
The second method involves the oxidation of the glucose to gluconic acid by a glucose oxidase. The oxygen used, proportional to the glucose present, is measured and expressed in percent glucose. Error similar to the o-toluidine method; time for analysis: 3 minutes per sample Imaximum)-Due to a combination of errors (error of the analysis of glucose plus error involved in the hydrolysis reaction) two results that differed from each other to an extent of 5% were considered identical.
Only in cases where the results were signifi-cantly different was it concluded that the parameters studied had an influence on the yield of glucose. r Urea was analyzed by hydrolyzing it to ammonia with the enzyme urease, and determining the ammonia either potentiometrically by the use of an ion specific electrode, or by colorimetry by reaction of the ammonia with sodium nitroprusside plus sodium salicylate in the presence of sodium dichloroisocyanurate.
Using this last method, the ammonia plus free urea present are analyzed together, and the final result expressed as percent of free urea present.
Ureides were determined by paper chromatography and by the use of a Waters liquid chromatograph, Model M6000*
using a system of columns packed with Bondapak X Corasil*, and as solvent, a mixture made up of four parts of a 2-to-1 mixture of isopropanol ethylacetate and one part distilled water.
Starch determination was done by treatment of *trade mark -:
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a known amount of barley suspended in water with 85% phos-phoric acid (pH 1,5 ~ 2) at 80C overnight. A control experiment was done at the same time with pure potato starch.
This first treatment liquefied all the starch present. After cooling and adjusting the pH to 4.5 sodium hydroxide, both unknown and control were treated with the same amount of glucoamylase at 62C for 24 hours. The glucose present was analyzed, and the result obtained from barley corrected according to the glucose obtained from the control experiment.
The content of starch was found to be 55% on a dry basis.
The Hydrolysis Reaction There are several known procedures available for hydrolyzing starch to produce glucose useful for this invention. For Qxample, one may liquify starch by mixing a 1 15 30% aqueous suspension of starch with 0.07% of ~-amylase at a pH of 6.9. The temperature is held at 70-75-80-85C for 15 minutes at each temperature. The mixture is then boiled ; for an hour, cooled to 85C, and 0.04% of ~-amylase is added - and the mixture kept at 82C for 30 minutes. Agitation is maintained throughout these steps. The reaction mixture is then cooled to 60C, the pH is adjusted to 4.0 - 4.5 with ; HCl, 0.1% of the enzyme amyloglucosidase (commonly termed glucoamylase) is added and the product incubated at 60C
for 24 hours to complete conversion of the starch to glucose.
However, we prefer to use a novel process wherein only one enzyme, glucoamylase, is used under particular conditions to convert at least 90% of the starch to glucose.
More particularly, an aqueous mash of ground barley is mixed with an appropriate amount of glucoamylase under acidic condition , 30 at temperatures within the range 65 - 67C and reacted for ; sufficient time to achieve the desired-degree of conversion.
i In a preferred embodiment, ground barley is slurried with water ln a proportion of between 140 - 240 parts per 100 parts 1()3~76~3 of grain. Acid~ usually 85% phosphoric acid, is added to bring the pH to about 3.0 ~ 4,5, Glucoamylase, preferably in the form of an aqueous solution containing 100 units per milliliter, is added and the temperature is then raised to about 66C. Eighty milliliters of enzyme solution will hydrolyze 100 pounds of liquified starch in 72 - ~6 hours at 62C and pH4. The enzyme addition may be from 30 to 60 units per 100 grams of barley. rrhe reaction is carried on for about 10 to 15 hours. A runny slurry or soup is obtained containing about 15% glucose. The process converts about 90~ of the starch to glucose.
A number of variables that could influence the course of the novel hydrolysis reaction have been inves-tigated. Each one was examined under a set of standard conditions, so only one variable was changed at a time. The ~- standard conditions were:
- 100 g. of whole barley, crushed to 4/64 inches - 240 ml. water - pH 4 - 4.5 by use of 85% phosphoric acid - 0.6 ml. of glucoamylase added in two batched;
0.3 ml. at zero time and 0.3 ml. four hourslQtQr - reaction time - 24 hours It was found that the ratio of 240 ml. of water/100 g. of barley gave a handleable porridge at the beginning of the reaction, and a final runny soup that was also easily handled. A pH close to 4 is the optimum acidity for the enzyme glucoamylase.
The following variables were found to have little effect on the reaction: particle size (ranges tried:
i~377~8 . ~ :
BACKGROUND OF THE INVENTION
This invention relates to a process for pro-ducing a non-protein nitrogen (NPN) containing material useful as a feed supplement for cattle and sheep. More ; 5 particularly, it relates to a process for producing urease-resistant glucosyl ureides and to the high assay product of the process.
It is now common to feed cattle and sheep feed supplements. These supplements provide nitrogen as a source for meeting the protein needs of the animals.
The most commonly used supplement is urea. When the urea enters the rumen of the animal, the enzyme urease acts on it to form ammonia. The microorganisms in the rumen then convert - the ammonia into a form of protein which can be utilized by - 15 the animal.
There is a disadvantage in using urea in that ; : , it is converted to ammonia at a relatively rapid rate. If the dosage of urea is too large, the capacity of the micro-organisms to convert the produced ammonia to protein is exceeded and the excess ammonia is either converted to urea and ex-creted by the animal or, if its concentration is high enough, ,, I_ , ~ .
,~
s;i, ~t can be tox~377~B
Recent work has been directed toward develop-ing "slow release" NPN compounds. The objective is to :.-provide compounds which produce ammonia in the rumen at a reduced rate with which the microorganisms can cope.
One family of compounds, glucosyl ureides, is known to have the property of slow release of ammonia in the rumen of cattle or sheep. These compounds have the added advantage of providing the ammonia in association with an energy source (glucose), which promotes greater growth of the microflora in the rumen. A United States patent of interest in this connection issued to McNeff under No. 3677767. This patent teaches reacting molasses (which contains glucose and non-reducing sugars, such as fructose) with urea under ~ 15 acidic conditions to produce a liquid product containing ; glucosyl ureide. The product has been shown to be useful as a feed supplement for ruminant animals.
Other pertinent prior art is exemplified by U.S. patents: 2612497, issued to Meijer; 3023205, issued to Meijer; and 3020273, issued to Steadman.
, ~
SUMMARY OF THE INVENTION
: `
.. ,, i The present invention is eoncerned with a - ! known reaction whereby glucose is eombined with urea under aeidie eonditions to produce glucosyl ureides. However, in aeeordance with the invention the process is carried out in a manner sueh that a relatively high degree of eonversion j of the glucose is aehieved, and the end product contains ¦ a relatively high concentration of ureides.
More particularly, glucose is reacted with urea under aeidie and drying eonditions to produee a produet, :' 1037~
.
preferably solid, containing a relatively hi~h proportion o~ urease - resistant glucosyl ureides.
- When first considering drying the glucose-urea Mixture during reaction to drive off water and shift the equilibrium of the reaction toward producing more ureides, we anticipated that the concentration of acid would increase, leading to dehydration and undesired carmelizing or degradation.
Surprisingly, the pH of the acidic mixture actually increases during the drying process. Carmelizing is not a serious problem.
In retrospect, it appears that ammonia, released from the urea, or protein, which is present if one uses hydrolyzed grain as the starting material, buffers the reaction to permit it to proceed to completion.
Since drying is an integral part of the process, we prefer to use a glucose-containing starting material which -is relatively free of other hexoses, which make drying difficult - and which can result in charring of the final product. We have found that grain starch, such as that occurring in barley, wheat or corn, may be hydrolyzed to provide a preferred starting material containing glucose. If, for example, molasses is used in the process, it will not readily dry to a useful product, due to the presence of a mixture of sugars and the tendency of contained fructose to char. However, we have found ; that some hygroscopic mixtures of sugars can be tolerated in the starting mixture, provided that inert materials, such as grain husks, are also present.
~- The product of the process comprises a solid, con-densation reaction product of (1) glucose-containing hydrolyzate of grain, selected from the group consisting of barley, wheat and corn, and (2) urea, in the presence of acid under drying condition at elevated temperature, said product containing the major propcrtion of said glucose in the form of glucosyl ureides.
.
~L~37!7~ :
The process for producing the desired mixture of glucosyl ureides comprises the steps of: providing an aqueous ~ ;
` mixture of a starting material containing one or more reducing carbohydrates as the major dry matter constituent, the major constituent of said reducing carbohydrates being glucose, and urea; and reacting the mixture under acidic and drying con~
ditions at an elevated temperature sufficient to convert the major portion of the glucose and urea into glucosyl ureides ~
and to produce a solid product~ ;
DESCRIPTION OF`THE DRAWINGS
` In the drawing:
' Figure 1 shows a preferred form of the process outlined in terms of chemical equations for the hydrolysis and condensation reactions; and Figure 2 is a plot showing the effect of tempera-' ture of the reactlon mixture on the hydrolysis reaction.
DESCRIPTION OF T~E PREFERRED E~BODIMENT
The invention will now be described in terms of providing glucose by hydrolyzing starch present in barley.
' 20 While a number of grains, such as wheat, corn and the like can be used, barley is preferred as it is commonly used in Western Canada as cattle feed. ' - - , Equipm_ t' _ d`analy's'is pro`c'edur`es The hydrolysis reaction was carried out in a ' ' :' 25 50 ml reactor provided with stirrer, inlets for introduction ' of reagents and sampling, a mantle, and a temperature controller ~'~
capable of keeping the temperature within'+ .5~C. ~ '' The glucose analyses were performed by the ; orthotoluidine method and by the use of a Beckman* ~lucose autoanalyzer, Model ERA-2001*. The first method involves the reaction of the glucose present with o-toluidine in acetic acid.
.. . .
~ ~ * trademark - 4 - '-. . ~.
: - . - ~, . :
..
Q37~
The color is measured spectrophotometrically using 5 determin-ations for each sample, one for the blank, and one for the standard. The error of the method was found to be 2%. Time ;
employed for each deter~in~tion: ~15 minutes.
The second method involves the oxidation of the glucose to gluconic acid by a glucose oxidase. The oxygen used, proportional to the glucose present, is measured and expressed in percent glucose. Error similar to the o-toluidine method; time for analysis: 3 minutes per sample Imaximum)-Due to a combination of errors (error of the analysis of glucose plus error involved in the hydrolysis reaction) two results that differed from each other to an extent of 5% were considered identical.
Only in cases where the results were signifi-cantly different was it concluded that the parameters studied had an influence on the yield of glucose. r Urea was analyzed by hydrolyzing it to ammonia with the enzyme urease, and determining the ammonia either potentiometrically by the use of an ion specific electrode, or by colorimetry by reaction of the ammonia with sodium nitroprusside plus sodium salicylate in the presence of sodium dichloroisocyanurate.
Using this last method, the ammonia plus free urea present are analyzed together, and the final result expressed as percent of free urea present.
Ureides were determined by paper chromatography and by the use of a Waters liquid chromatograph, Model M6000*
using a system of columns packed with Bondapak X Corasil*, and as solvent, a mixture made up of four parts of a 2-to-1 mixture of isopropanol ethylacetate and one part distilled water.
Starch determination was done by treatment of *trade mark -:
,~
~037 ~ ~
a known amount of barley suspended in water with 85% phos-phoric acid (pH 1,5 ~ 2) at 80C overnight. A control experiment was done at the same time with pure potato starch.
This first treatment liquefied all the starch present. After cooling and adjusting the pH to 4.5 sodium hydroxide, both unknown and control were treated with the same amount of glucoamylase at 62C for 24 hours. The glucose present was analyzed, and the result obtained from barley corrected according to the glucose obtained from the control experiment.
The content of starch was found to be 55% on a dry basis.
The Hydrolysis Reaction There are several known procedures available for hydrolyzing starch to produce glucose useful for this invention. For Qxample, one may liquify starch by mixing a 1 15 30% aqueous suspension of starch with 0.07% of ~-amylase at a pH of 6.9. The temperature is held at 70-75-80-85C for 15 minutes at each temperature. The mixture is then boiled ; for an hour, cooled to 85C, and 0.04% of ~-amylase is added - and the mixture kept at 82C for 30 minutes. Agitation is maintained throughout these steps. The reaction mixture is then cooled to 60C, the pH is adjusted to 4.0 - 4.5 with ; HCl, 0.1% of the enzyme amyloglucosidase (commonly termed glucoamylase) is added and the product incubated at 60C
for 24 hours to complete conversion of the starch to glucose.
However, we prefer to use a novel process wherein only one enzyme, glucoamylase, is used under particular conditions to convert at least 90% of the starch to glucose.
More particularly, an aqueous mash of ground barley is mixed with an appropriate amount of glucoamylase under acidic condition , 30 at temperatures within the range 65 - 67C and reacted for ; sufficient time to achieve the desired-degree of conversion.
i In a preferred embodiment, ground barley is slurried with water ln a proportion of between 140 - 240 parts per 100 parts 1()3~76~3 of grain. Acid~ usually 85% phosphoric acid, is added to bring the pH to about 3.0 ~ 4,5, Glucoamylase, preferably in the form of an aqueous solution containing 100 units per milliliter, is added and the temperature is then raised to about 66C. Eighty milliliters of enzyme solution will hydrolyze 100 pounds of liquified starch in 72 - ~6 hours at 62C and pH4. The enzyme addition may be from 30 to 60 units per 100 grams of barley. rrhe reaction is carried on for about 10 to 15 hours. A runny slurry or soup is obtained containing about 15% glucose. The process converts about 90~ of the starch to glucose.
A number of variables that could influence the course of the novel hydrolysis reaction have been inves-tigated. Each one was examined under a set of standard conditions, so only one variable was changed at a time. The ~- standard conditions were:
- 100 g. of whole barley, crushed to 4/64 inches - 240 ml. water - pH 4 - 4.5 by use of 85% phosphoric acid - 0.6 ml. of glucoamylase added in two batched;
0.3 ml. at zero time and 0.3 ml. four hourslQtQr - reaction time - 24 hours It was found that the ratio of 240 ml. of water/100 g. of barley gave a handleable porridge at the beginning of the reaction, and a final runny soup that was also easily handled. A pH close to 4 is the optimum acidity for the enzyme glucoamylase.
The following variables were found to have little effect on the reaction: particle size (ranges tried:
2/64 - 4/64 inches); type of water (tap water vs. distilled ¦ water); stirring; whole or dehusked ~arley; type of barley;
! moisture content of the barley; and temperature gradient within the reactor.
~ ~03776~
One variable, temperature of the reaction ; mixture, was found to have an effect on glucose yield.
As illustrated in Figure 2, glucose yields were poor outside the reaction temperature range of 65 - 67C. Yields were maximized at about 90% within this range.
The process has advantages over the prior art in that (1) it is a single enzyme treatment as compared to the double enzyme treatments previously usedi (2) lower temperatures are used; and (3) the reaction does not require vigorous agitation. These differences result in a simpler process.
- The Condensation Reaction Once the glucose - containing hydrolyzate is available, it is reacted with urea under acidic drying conditions to produce gluccsyl ureides. The reaction is preferably carried out to maximize the yield of ureides, ;
particularly of monoglucosyl ureide, while minimizing the production of caramel and other products of the decomposition of glucose. Therefore it is normally conducted under mild temperature conditions to produce a solid product. At this time, the process has been developed to the point where yields 1, in the order of 90% may be achieved.
The hydrolyzate is reacted with an amount of urea which is preferably slightly in excess of that amount required stoichiometrically to combine with the glucose to produce monoglucosyl ureide. We normally use about a 10%
excess of urea. One can, if desired, react the glucose with less than the stoichiometric amount - this approach has the advantage of providing free glucose in the supplement, thereby improving its palatability; however, normally the objective of maximizing conversion to ureides is overiding.
Alternatively, one can use a larger excess than 10%. However, .
~l037'~'6~
we have not noted any significant increase in yield when excesses of 200% and 300% were used.
The pH of the starting reaction mixture is ; established at about 2 - 4.
The drying operation is influenced by ~ temperature, retention time and depth of bed. In general,I we seek to maximize conversion by drying to a solid state ';' ' without sianificant charring. One should keep in mind that, if the temperature is high, charring can occur if the retention time is too long. Most of our drying investigations have been carried out on a laboratory batch basis using a thin layer (about 5/16") of reaction mixture on trays in a hot air oven. Under these conditions, drying is carried out at a temperature of about 75-90C for a prolonged periodj ~ 15 usually in the order of 10 - 24 hours. However, we have .. ' I
also successfully carried out trials in a rotary hot air drier having an inlet air temperature of 600F and outlet temperature of 300F. The retention time used was only .,.,. ~
I seconds to produce a solid product.
In the case of barley, the preferred end product is a solid brown material having the following typical composition:
monoglucosyl ureide - 21.0%
diglucosyl ureide - 8.0%
Water insoluble matter from barley - 25.0%
Water soluble matter from barley - 16.0%
Free glucose - 3.5%
Free urea - 4.4%
Phosphate - 4.5%
Water - 3.0%
Unidentified compounds - 14.6%
(Mostly nitrogenous compounds derived from glucose and urea) :
'' 103776~
The invention will be further understood ; with reference to the following examples:
Example 1 This example provides a detailed description of a preferred method for carrying out the hydrolysis and condensation reactions.
.. .. .
The example involved hydrolysis of whole barley, followed by the reaction of said hydrolyzed barley ; with urea plus phosphoric acid. lOOg of crushed whole barley with a moisture content of 11%, 240 ml of water, and 0.6g I of 85% phosphoric acid were mixed together at room temperature.
Thirty units of Diazyme L-1003 were added to the mixture and heat was applied with stirring so that the reaction mixture was brought to 65C in approximately 45 minutes. Thre~ hours after having reached this temperature, 30 more units of Diazyme L-100 were added and the mixture kept for approximately 10 hours at 65C under stirring.
At this stage, the glucose content of the barley hydrolyzate was found to be 16.3~ of 90~ of the theoretical amount.
To lOOg of barley hydrolyzate containing 14g I of glucose were added 4.68g of urea (no excess) and lg of 85% phosphoric acid. The mixture was spread out on a plastic tray, the thickness of said mixture being 6 to 7 mm, and held at 75C for 14 hours. At this time analysis of glucose by the glucose oxidase method showed 5.5%, which represents a conversion of 89%. Analysis of the product showed urea plus ammonia, expressed as urea, 3.36%; ureides expressed as monoglucosyl urea, 37.1~4; combined nitrogen, expressed as monoglucosyl urea, 37~; total nitrogen, 7.76~.
3Registered trade mark for glucoamylase manufactured by Miles Laboratories, Elkhart, Indiana, U.S.A. Eighty units of said enzyme will hydrolyze 100 pounds of liquefied starch in 72 to 96 hours at 62C and pH4.
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4A~alyzed by a potentiometric method.
Example II
This example shows the results obtained when a 10% excess of urea is used.
To lOOg of barley hydrolyzate prepared according to Example I, containing 15.6g of glucose, were added 5.7g of urea (10% excess) plus l.lg of 85% phosphoric acid. The mixture was placed on a plastic tray, the thickness of said mixture being close to 7 mm, and kept at 75C for 14 hours. At this stage, the solid product showed the following analysis: Glucose, 1% (conversion of glucose 98%); ammonia, expressed as urea, 0.46%; ammonia plus free urea, expressed as urea, 3.74%; ureides, expressed as monoglucosyl urea, 40.3%4; combined nitrogen, expressed as monoglucosyl urea, 43%; total nitrogen, 8.6%.
;' Example III
This example shows the effect of increasing the temperature during the drying-operation.
' To lOOg of barley hydrolyzate, prepared " 20 according to Example I and containing 12.6g of glucose, were added 4.6g of urea (10% excess) plus lg of 85% phosphoric ' acid. The mixture was placed on a plastic tray, the thickness of said'mixture being 10 mm, and kept at 95C for 5 hours.
At this time the solid material was broken into small pieces and held at 95C for 2 more hours. The solid material gave ~' the following analysis: glucose, 2% -(conversion of glucose 94~); ammonia, expressed as urea, 0.26%; ammonia plus free urea, expressed as urea, 2.9%; ureides, expressed as monoglucosyl urea, 34.4~4; combined nitrogen, expressed as monoglucosyl urea, ' 30 40%; total nitrogen, 8.0%.
Ex-ample''IV
I This example is similar to Example II, however .. I .
-- 1 1 -- ' , 776~
a longer retentid~ tlme is used, To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 4.76g of urea (10~ excess~ plus 0.95g of 85% phosphoric acid. lOOg of this mixture were poured into a plastic tray, the thickness of the said mixture being 7 mm, and kept at 75C for 24 hours. At this stage, 30.6g of solid material were obtained that gave the following analysis: glucose,
! moisture content of the barley; and temperature gradient within the reactor.
~ ~03776~
One variable, temperature of the reaction ; mixture, was found to have an effect on glucose yield.
As illustrated in Figure 2, glucose yields were poor outside the reaction temperature range of 65 - 67C. Yields were maximized at about 90% within this range.
The process has advantages over the prior art in that (1) it is a single enzyme treatment as compared to the double enzyme treatments previously usedi (2) lower temperatures are used; and (3) the reaction does not require vigorous agitation. These differences result in a simpler process.
- The Condensation Reaction Once the glucose - containing hydrolyzate is available, it is reacted with urea under acidic drying conditions to produce gluccsyl ureides. The reaction is preferably carried out to maximize the yield of ureides, ;
particularly of monoglucosyl ureide, while minimizing the production of caramel and other products of the decomposition of glucose. Therefore it is normally conducted under mild temperature conditions to produce a solid product. At this time, the process has been developed to the point where yields 1, in the order of 90% may be achieved.
The hydrolyzate is reacted with an amount of urea which is preferably slightly in excess of that amount required stoichiometrically to combine with the glucose to produce monoglucosyl ureide. We normally use about a 10%
excess of urea. One can, if desired, react the glucose with less than the stoichiometric amount - this approach has the advantage of providing free glucose in the supplement, thereby improving its palatability; however, normally the objective of maximizing conversion to ureides is overiding.
Alternatively, one can use a larger excess than 10%. However, .
~l037'~'6~
we have not noted any significant increase in yield when excesses of 200% and 300% were used.
The pH of the starting reaction mixture is ; established at about 2 - 4.
The drying operation is influenced by ~ temperature, retention time and depth of bed. In general,I we seek to maximize conversion by drying to a solid state ';' ' without sianificant charring. One should keep in mind that, if the temperature is high, charring can occur if the retention time is too long. Most of our drying investigations have been carried out on a laboratory batch basis using a thin layer (about 5/16") of reaction mixture on trays in a hot air oven. Under these conditions, drying is carried out at a temperature of about 75-90C for a prolonged periodj ~ 15 usually in the order of 10 - 24 hours. However, we have .. ' I
also successfully carried out trials in a rotary hot air drier having an inlet air temperature of 600F and outlet temperature of 300F. The retention time used was only .,.,. ~
I seconds to produce a solid product.
In the case of barley, the preferred end product is a solid brown material having the following typical composition:
monoglucosyl ureide - 21.0%
diglucosyl ureide - 8.0%
Water insoluble matter from barley - 25.0%
Water soluble matter from barley - 16.0%
Free glucose - 3.5%
Free urea - 4.4%
Phosphate - 4.5%
Water - 3.0%
Unidentified compounds - 14.6%
(Mostly nitrogenous compounds derived from glucose and urea) :
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The invention will be further understood ; with reference to the following examples:
Example 1 This example provides a detailed description of a preferred method for carrying out the hydrolysis and condensation reactions.
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The example involved hydrolysis of whole barley, followed by the reaction of said hydrolyzed barley ; with urea plus phosphoric acid. lOOg of crushed whole barley with a moisture content of 11%, 240 ml of water, and 0.6g I of 85% phosphoric acid were mixed together at room temperature.
Thirty units of Diazyme L-1003 were added to the mixture and heat was applied with stirring so that the reaction mixture was brought to 65C in approximately 45 minutes. Thre~ hours after having reached this temperature, 30 more units of Diazyme L-100 were added and the mixture kept for approximately 10 hours at 65C under stirring.
At this stage, the glucose content of the barley hydrolyzate was found to be 16.3~ of 90~ of the theoretical amount.
To lOOg of barley hydrolyzate containing 14g I of glucose were added 4.68g of urea (no excess) and lg of 85% phosphoric acid. The mixture was spread out on a plastic tray, the thickness of said mixture being 6 to 7 mm, and held at 75C for 14 hours. At this time analysis of glucose by the glucose oxidase method showed 5.5%, which represents a conversion of 89%. Analysis of the product showed urea plus ammonia, expressed as urea, 3.36%; ureides expressed as monoglucosyl urea, 37.1~4; combined nitrogen, expressed as monoglucosyl urea, 37~; total nitrogen, 7.76~.
3Registered trade mark for glucoamylase manufactured by Miles Laboratories, Elkhart, Indiana, U.S.A. Eighty units of said enzyme will hydrolyze 100 pounds of liquefied starch in 72 to 96 hours at 62C and pH4.
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4A~alyzed by a potentiometric method.
Example II
This example shows the results obtained when a 10% excess of urea is used.
To lOOg of barley hydrolyzate prepared according to Example I, containing 15.6g of glucose, were added 5.7g of urea (10% excess) plus l.lg of 85% phosphoric acid. The mixture was placed on a plastic tray, the thickness of said mixture being close to 7 mm, and kept at 75C for 14 hours. At this stage, the solid product showed the following analysis: Glucose, 1% (conversion of glucose 98%); ammonia, expressed as urea, 0.46%; ammonia plus free urea, expressed as urea, 3.74%; ureides, expressed as monoglucosyl urea, 40.3%4; combined nitrogen, expressed as monoglucosyl urea, 43%; total nitrogen, 8.6%.
;' Example III
This example shows the effect of increasing the temperature during the drying-operation.
' To lOOg of barley hydrolyzate, prepared " 20 according to Example I and containing 12.6g of glucose, were added 4.6g of urea (10% excess) plus lg of 85% phosphoric ' acid. The mixture was placed on a plastic tray, the thickness of said'mixture being 10 mm, and kept at 95C for 5 hours.
At this time the solid material was broken into small pieces and held at 95C for 2 more hours. The solid material gave ~' the following analysis: glucose, 2% -(conversion of glucose 94~); ammonia, expressed as urea, 0.26%; ammonia plus free urea, expressed as urea, 2.9%; ureides, expressed as monoglucosyl urea, 34.4~4; combined nitrogen, expressed as monoglucosyl urea, ' 30 40%; total nitrogen, 8.0%.
Ex-ample''IV
I This example is similar to Example II, however .. I .
-- 1 1 -- ' , 776~
a longer retentid~ tlme is used, To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 4.76g of urea (10~ excess~ plus 0.95g of 85% phosphoric acid. lOOg of this mixture were poured into a plastic tray, the thickness of the said mixture being 7 mm, and kept at 75C for 24 hours. At this stage, 30.6g of solid material were obtained that gave the following analysis: glucose,
- 3.2% (conversion of glucose 92%); ammonia plus urea, 2.81%
monoglucosyl urea, 18.9%; combined nitrogen, expressed as monoglucosyl urea, 46~; total nitrogen, 8.67~.
- Example V
This example shows the use of only 85% of the stoichiometric amount of urea required to react with the glucose to produce monoglucosyl ureide.
To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 3.68g of urea (85% of stoichiometric amount) plus 0.73g ; of 85% phosphoric acid. lOOg of this mixture were poured into a plastic tray, the thickness of the said mixture being 7mm, and kept at 75C for 24 hours. At this time, 30g of solid material were obtained that gave the following analysis:
glucose, 8.1% (conversion of glucose 80%); ammonia plus urea, expressed as urea, 2.06%; monoglucosyl urea, 16.7%; combined nitrogen, expressed as monoglucosyl urea, 36%; total nitrogen, 7.0%.
Example VI
This example shows the effect of using a 100%
' excess of urea.
To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 8.66g of urea (100% excess) plus 1.73g of 85% phosphoric acid. lOOg ':
.
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of this mixture were poured into a plastic tray, the thickness of the said mixture being 7 mm, and kept at 75C for 24 hours.
At this stage, 32.9g of solid material were obtained that ~ gave the following analysis: glucose, 0.6~ (conversion of - 5 glucose 98.3~); ammonia plus urea,~expressed as urea, 10.67%
, : monoglucosyl urea, 27.~%; combined nitrogen, expressed as - monoglucosyl urea, 47%; total nitrogen, 12.27%.
Example VII
- This example shows the use of corn as the source of starch in the hydrolysis reaction.
lOOg of corn (whole grain, 18.9% moisture) were mixed with 240 mls of tap water and ground exhaustively.
The pH was adjusted to 4.5 with 85~ phosphoric acid and 30 units of Diazyme L-100 added. Stirring was commenced and the temperature was raised to 67C. At this time, 30 units of Diazyme L-100 were added and the reaction was carried on for 24 hours. A yellow, runny soup was obtained - it contained 14.6% glucose.
Example VIII
This example shows the use of wheat as the source of starch in the hydrolysis reaction.
lOOg of whole ground wheat (12.4% moisture) were mixed with 240 mls of tap water. The pH was adjusted to 4.7 by the addition of 0.6g of 85~ phosphoric acid. 30 units of Diazyme L-100 were added and stirring was commenced.
The temperature was raised to 67C over a period of 1 hour.
The reaction was continued at this temperature and, after i 3 hours, an additional 30 units of Diazyme L-100 were added.
The reaction was then continued for a total reaction time of 16 hours. 340 g of hydrolyzate containing 14% glucose were - obtained.
To lOOg of the wheat hydrolyzate were added : . . . .
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: ~ 10377f~
5.2g of urea (10% excess) plus 1~ of 85% phosphoric acid.
The mixture was placed in a plastic tray to a thickness of 7 mm, and kept at 75C for 14 hours. At this stage, the solid material gave the following analysis: glucose, 4.8%
(conversion 90%); ammonia plus free urea, expressed as urea, 2.39~; ureides, 4 expressed as monoglucosyl urea, 41.63%;
total nitrogen, 8%.
Example IX
This example shows the results of drum-drying.
Barley hydrolyzate containing 13.8~ glucose was rapidly evaporated using a rotary drum under the following conditions:
Thickness of product: 0.025 inches Steam pressure: 48 psi :
Production: 1.45 pounds of dry material per foot 2 per hour ` Slurry feed rate: 4.55 pounds/hour Surface area: 0.962 foot rying time: 37 seconds --In this way, a rapid evaporation of water was ' 20 achieved, however less glucose was converted to ureides - ! compared to the previous examples. A solid product was - obtained that gave the following analysis: glucose, 15%
(conversion 70%); urea plus ammonia, expressed as urea, 5.41%; combined nitrogen, expressed as monoglucosyl urea, 35.0%; monoglucosyl urea, 12.7%; total nitrogen, 8.4%.
! Example X
:., I
This example shows the results of a centrifuge and evaporation trial.
200 g of barley hydrolyzate (13% glucose) plus 10~ excess urea were centrifuged at 200 rpm for 20 hours.
105g of liquid and 105g of cake were obtained. The liquid and the cake were evaporated to yieId 45.15 g of syrup, ,. I
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and 55.?g of cake, respectively; the~ were mixed together to give a mixture with a water content of 35%. This mixture was dried at 75C for 20 hours. 73g of a solid were obtained - that gave the following analysis: glucose, 3% (conversion 90%);
ammonia plus urea, expressed as urea, 3.4%; combined nitrogen, expressed as monoglucosyl urea, 37%, total nitrogen, 8.51%.
Example XI
This example shows the results obtained when the hydrolyzate was cooked instead of dried in accordance with the invention To lOOOg of barley hydrolyzate containing 138g of glucose, were added 50g of urea (10% excess) plus lOg of 85% phosphoric acid. lOOOg of this mixture were evaporated to ; 395g of solid. This solid was placed in a closed container and kept at 75C for 14 hours. At this stage no loss of water was observed. The solid pastry product showed the following analysis: glucose, 8% (conversion of glucose 77%); urea plus ammonia, expressed as urea, 3.98%; combined nitrogen, expressed as monoglucosyl urea, 25%; total nitrogen, 6.76%.
A principal advantage of our product is that it can be produced with a high total equivalent crude protein ` content, with up to 90% of this crude protein in a slow ... .
ammonia release, urease-resistant form. The following examples are from animal feeding trials using various forms of our product and demonstrating the effectiveness of the product.
Example XII
For the purpose of this trial, steers of Here-ford breeding were divided into two groups; one group fed a ration which contained no supplemental protein, the other group fed a ration which contained the solid NPN product prepared as in Example IV. These mixtures constltuted 33%
of the final ration and were formulated to contain similar '~ ..
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.~
levels of all nutrients except crude protein. See Table 1.
- Formulat1on of Diets Ingredients Control Solid non-protein ration nitrogen ration (g~) (%) ~ ,: ~ ' . .. .
Straw (oats) 66.7 66.7 Concentrate 33.3 33.3 Composition of Concentrate Barley 93.7 61.5 - Ground limestone 1.6 2.4 Trace mineralized salt 1.0 1.0 Vitamin premix 11 2.0 2.2 Barley-based non-protein nitrogen -- 32.8 To supply 2,300, 380 and 2.3 IU of Vitamins A, D3 and E, respectively, per pound of total ration.
Compo_ltion of Total Ration Moisture ................. (%) 15.8 15.5 Crude protein............. (%) 5.9 8.9 - Calcium................... (%) 0.3 0.34 Phosphorus................ (~) 0.21 0.26 Digestible energy......... Mcal/pound 1.08 1.05 From Table 2, we see that the steers fed with the solid non-protein nitrogen supplement gained 20% faster than did the animals that received the control ration; they also ate more and needed a lower amount of intake per pound gained.
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T~BLE 2 Effects of Crude Protein Supplementation On the Perfoxmance of Growing Steers - Control Non-protein Nitrogen Number of steers 16 16 Feeding period days 154 154 Av. initial weight pounds 527 530 Av. final weight pounds 612 632 Av. daily gain pounds 0.55 0.66 Av. daily feed pounds 12.5 13.3 Feed/gain 22.47 21.2 . _ _ 12Feed intake is given on an as-fed basis (approximately 84.5%
dry matter).
Example XIII
In this example, three groups of steers were fed rations supplemented with one of the following: (l)our NPN
product, prepared as in example IV; (2)soymeal (which is a premium plant source of protein); and (3)urea. A fourth control~
group was not fed any supplement.
The growing ration used was based on corn silage with a "barley-protein supplemented" concentrate to provide an overall ration with an energy equivalent of approximately 1.17 Mcal per pound of dry matter. The equivalent crude protein content of the unsupplemented control was 0.94 pounds per pound of dry matter, while that of the supplemented rations was 0.11 pounds per pound of dry matter.
The results of the trial, the compositions of the concentrates, and the nutrient content of the feed are given in the following tables:
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1()37768 (Control)(Soymeal)(Test NPN)(Urea) (No supplement) Number of animals 144 143144 144 Days on test 118 105103 119 Pounds Silage consumed per head/day 47.89 50.27 49.30 48.97 Concentrate consumed per head/day 2.99 3.13 3.07 3.06 Pounds total dry matter consumed per head/day14.66 15.39 15.09 14.99 Average total gain (pounds per head)184.7 185.7184.5195.4 Average daily gain1.56 1.77 1.79 1.64 - Conversion ratio (dry matter consumed9.37 8.70 8.43 9.13 per pound gain) ~. . _ . __ _ TABLE IV
Concentrate Composition : .
Low Soybean Test Portein Urea Meal NPN
:........................................ _ ~ Ingredient:
;: 25 Ground Barley 95.35 92.1271.17 74.01 Urea (45% N) -- 3.23 -- --Soybean Meal -- --24.58 --Test NPN -- -- -- 21.56 Dicalcium phosphate -- 0.06 -- ---30 Ground Limestone 2.00 1.941.84 1.95 Plain Salt 1.32 1.321.35 1.86 Beef Mineral/Vitamin Pre. 0.332 0.332 0.332 0.332 Sodium tripolyphosphate 0.710 0.710 0.430 --Elemental Sulfur0.286 0.286 0.286 0.286 - Total 100.0 100.0100.0 100.0 Nutrient:
Dry Matter (%) 88.56 88.9588.78 88.96 ' Crude Protein (%)11.08 19.7719.75 19.78 i Digestible Protein (%) 6.88 -- 15.47 __ ,-40 M.E. Rum. (Mcal/lb)1.144 1.106 1.115 1.101 i TDN (%) 69.56 67.2069.87 66.90 Calcium (%) 0.810 0.797 0.813 0.807 Phosphorus (%) 0.515 0.515 0.519 0.515 .
.
lV37~6~3 Sodium (%) 0.749 0.749 0.752 0.761 Magnesium (%) 0.092 0.089 0.135 0.096 Potassium (%~ 0.361 0.349 0.740 0.375 Sulfur (%) 0.481 0.477 0.554 0.481 Iron (mg/lb) 46.9 46.1 56.0 54.4 Copper (mg/lb) 14.5 14.4 15.5 30.5 Manganese (mg/lb)22.4 22.2 24.0 22.7 Zinc (mg/lb) 78.2 77.6 74.3 85.2 Crude Fat (%) 1.8 1.8 1.7 1.4 Crude Fiber (%) 4.8 4.6 4.9 3.7 TABLE V
Nutrient Content of a 1:4- Blend of Concentrate and Corn Silage C.M. (% of DM) Low Soybean Test Protein Urea Meal NPN
----___ _ _ __ Dry Matter (%) 100.0 100.0 100.0 100.0 Crude Protei~ (%)9.43 11.20 11.20 11.20 M.E. (Rum) (Mcal/lb)* 1.176 1.167 1.177 1.166 ;
TDN (%)* 71.55 71.01 71.60 70.95 Calcium (%) 0.379 0.376 0.379 0.377 '-. Phosphorus (%) 0.335 0.334 0.335 0.334 Sodium (%)- 0.211 0.210 0.211 0.213 Magnesium (%) 0.207 0.206 0.175 0.187 Potassium (%) 0.934 0.934 1.01 0.936 Sulfur (%) 0.123 0.122 0.138 0.123 Iron (mg/lb) 115.6 115.40 117.5 117.0 Copper (mg/lb) 5.9 5.9 6.1 9.2 Manganese (mg/lb)20.6 20.5 20.9 20.6 Zinc (mg/lb) 30.5 30.4 29.7 31.9 Fiber (Mg/lb) 22.7 22.7 22.7 22.5 Vitamin A (I.U./lb)3700.03700.03700.0 3700.0 .
; ME - Metabolitable energy TDN - Total digestible nutrients From these data, it will be seen that:
(1) in terms of feed conversion ratio, the . - 19 -, -~- 1~76~
the NPN supplement out-performed all other groups, followed by the soymeal, the urea, ancl the control group. The :: ;
observed difference betw~en the soymeal and the NPN group - and other groups -was statistically significant at the 95%
; confidence level, while the difference between the soymeal and the NPN group was not significant at this level.
(2) the soymeal and the NPN groups gained ~: .
approximately at the same rate, and at a significantly higher rate than the control or the urea group. The urea -; group gained at a significantly faster rate ` 15 than the control group.
(3) in terms of feed intake, the soymeal group consumed significantly more, while the control group consumed significantly ~` less feed than the other groups.
While certain examples~ structures~ composi-tions and process steps have been described for purposes of - illustration the invention is not limited to these. Varia~
tions and modifications within the scope of the disclosure and the claims can readily be effected by those skilled in the art.
SUPPLEM~NTA~Y PISCLOSURE
This supplementary disclosure relates to the use of starch-containing materials other than grains selected from the group consisting of barley, wheat and corn in the process of the Prîncipal Disclosure for producing a non-protein nitrogen-containing material useful as a feed supplement for cattle and sheep. The starch-containing materials may ~5 ~37~6~
i include different grains, such as rice and oats, as well as sources other than grains, such as cassava roots and potatoes.
; The characteristics of various suitable starch sources are given in Table 7.
TAsLE 7 Starch ~oisture Starch Source (%) - (%, d.m.b.) barley 10.8 59.9 sorghum 8.5 57.9 10 tapioca 10.0 96.6 rice 11.9 95.6 oats 10.5 53.3 : corn 9.8 82.1 cassava root 7.6 93.7 15 potatoes 78.4 69.5 The invention is not restricted to materials which hydrolyze to produce glucose as the major hydrolysis product. Glycoses, HOCH2 ~CHO~)nCHO, other than glucose can be ~ -reacted with urea to produce glycosyl ureides. For example, the spent sulphite liquours of the paper industry containxylose in amounts greater than glucose and can be used in the process of the present invention.
Broadly stated, the invention is a process for producing a non-protein nitrogen feed supplement for ruminants which comprises: providing an aqueous mixture of a starting - material containing one or more reducing carbohydrates as the major dry matter constituent, and urea; and reacting the mixture under acidic and drying conditions at an elevated temperature sufficient to convert the major portion of the reducing carbohydrates and urea into ureides and to produce a solid product.
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ANALYS I S` PROCEDURES
, Analysis procedures similar to those described in the Principal Disclosure were used unless otherwise specified.
The concentration of total ureides was deter-mined by acid hydrolysis of the product to give glucose and urea, followed by enzymatic hydrolysis of the urea and by the ~- potentiometric measurement of the resulting ammonia. The -`, measurement gave the total ammonia concentration; this value was corrected for the amount of free ammonia and free urea to ~,. 1 ~X` 10 obtain the total ureide concentration.
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The moisture content of a starch source was determined by heating a sample in an oven at 75C to constant weight.
Starch determination was done by treatment of a known amount of the starcll source suspended in water with ; 85% phosphoric acid (pH 1.5 - 2.0) at 80C overnight. A
control experlment was simultaneously conducted using pure potato staxch. After cooling and adjusting the p~I to 4.5 with sodium hydroxide, the unknown and the control samples were treated with the same amount of glucoamylase at 62C for 24 hours. The glucose thus produced was analyzed, and the result obtained was corrected according to the glucose obtained from the control experlment.
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THE HYDROLYSIS REACTION
The hydrolysis reaction was carried out in equipment similar to that described in the Principal Disclosure.
The standard reaction conditions were~
100 g substrate, crushed to 2/64"
24Q ml distilled water pH 4.0 to 4.5 (by use of 85% phosphoric acid) 1.0 ml Diazyme L-100* added in two batches;
0.5 ml at zero time and 0.5 ml three hours after reaction temperature reached 65C
reaction time: 24 hours The relatively low conversions to glucose in the case of sorghum and corn prompted the pre-treatment of the substrate at high temperature and acidic conditions overnight, prior to hydrolysis. In all cases, this procedure substantially increased the conversion of starch to glucose.
It was also found that the mixing of the sub-strate with water in a ~aring* Blender prior to the hydrolysis reaction also increased thé conversion to glucose. This was tried in the case of sorghum, corn, and potatoes.
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~ *trademark - 22 -~S~3~'~6~
- Due to the high moisture content, potatoes were the only starch source which was not crushed prior to reaction.
Instead, the potatoes were cut into small pieces with a knife and mixed with water in a Waring* slender. The soup obtained after hydrolysis was extremely runny, indicating that far less than the standard 240 mls of water could be used. A
reaction using only 120 mls of water was tried; this also pro-duced a very runny soup.
- The standard reaction time of 24 hours was cut to 6 hours when it was discovered that ~he heater malfunctioned overnight. During the 6-hour reaction it was observed that the controller maintained the temperature constant to within - 1.0C. The average increase in glucose content between 6 hours of reaction and 24 hours of reaction was 2.2~ based on analysis of the first 13 hydrolyses in which the heater was thought to have Punctioned properly.
THE CONDENSATION REACTION
The condensation reaction was carried out by mixing the hydrolyzate with a lQ~ stoichiometric excess of urea and with phosphoric acid ta obtain a pH of 2.5 - 3Ø The mixture was spread on a plastic tray and heated at a given temperature to dryness. The product was analyzed for glucose and monoglucosyl ureides.
The following examples illustrate the use of various starch-containing materials in the process of the present lnvention.
Example 14 This example shows the use of sorghum as the source of starch in the hydrolysis reaction.
: 30 10-Q g of crushed sorghum were mixed with 240 mls of distilled water. The pH was adjusted to 2.0 with 85~ -~ .
* trademark - 23 -~ ()37~
phosphoric acid and the mixture was left at 80C overnight.
After cooling and adjusting the pH to 4.5 with sodium hydroxide, :.
50 units of Diazyme L-100* were added. Stirring was commenced and the temperature was raised to 65C. Three hours a~ter having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 15%.
5.5 g of urea (a 10% excess) and 2 g of 85%
; 10 phosphoric acid were added to 100 g of the sorghum hydrolyzate.
The mixture was spread in a plastic tray to a thickness of 7 mm, ~ ~ -and kept at 75C for 2a hours. At this stage~ the solid -material gave the following analysis: glucose, 3% (93% con~
version); ammonia, expressed as urea, 0.44%; ammonia plus free -urea, expressed as urea, 5.6%; ureides, expressed as mono-glucosyl urea, 34.8% com~ined nitrogen, expressed as mono-glucosyl urea, 42~1%; total nitrogen 8.51%.
Example 15 -; This example shows the use of oats as the source of starch in the hydrolysis reaction.
100 g o~ crushed oats were mixed with 24Q mls ofdistilled water. The pH was adjusted to 4.5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 5Q units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the h~drol~zate was found to be 15.g%.
5.84 g of urea (a 10% excess) and 2 g of 85%
phosphoric ac~d were added to 100 g of the oat hydrolyzate.
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~ ~ * trademark - 24 -..
7~ 3 The mixture was spread in a plastic tray to a thickness of 7mm, and kept at 75C for 70 hours. At this stage, the solid material gave the following analysis: glucose, 1~ (98% con-version); ammonia, expressed as urea, 0.58%; ammonia plus free urea, expressed as urea, 5.0%; ureides, expressed as mono-glucosyl urea, 38.9%; combined nitrogen, expressed as mono-glucosyl urea, 45.0%; total nitrogen, 9.52%.
Example 16 This example shows the use of rice as the source of starch in the hydrolysis reaction.
lQ0 g of crushed brown rice were mixed with 240 mls of distilled water. The pH was adjusted to 4~5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
Stirring was commenced and the temperature was raised to 65C.
Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 21 4%
- 20 7~84 g o~ urea (lQ% excess) and 3 g of 85% -phosphoric acid were added to 100 g of the rice hydrolyzate.
The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 75~C for 50 hours. At thîs stage, the solid material gave the following analysis: glucose, 1% (98% con-version~; ammonia, expressed as urea, 0.78%; ammonia plus free urea, expressed as urea, 5.5%; ureides, expressed as mono-; glucosyl urea, 37.0%; combined nitrogen, expressed as mono-;~ glucosyl urea, 61.5%; total nitrogen, 11.3%.
Example 17 . '.
` 30 This example shows the use of corn as the source of starch in the hydrolysis reaction.
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lQQ g of crushed corn were mixed with 240 mls ~ -of distilled water and ground exhaustively. The pH was ad-justed to 4.5 with 85% phosphoric acid and 50 units of Diazyme L-100* were added. Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 6 hours, at which stage the glucose content of the hydrolyzate was found to be 10.8%.
monoglucosyl urea, 18.9%; combined nitrogen, expressed as monoglucosyl urea, 46~; total nitrogen, 8.67~.
- Example V
This example shows the use of only 85% of the stoichiometric amount of urea required to react with the glucose to produce monoglucosyl ureide.
To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 3.68g of urea (85% of stoichiometric amount) plus 0.73g ; of 85% phosphoric acid. lOOg of this mixture were poured into a plastic tray, the thickness of the said mixture being 7mm, and kept at 75C for 24 hours. At this time, 30g of solid material were obtained that gave the following analysis:
glucose, 8.1% (conversion of glucose 80%); ammonia plus urea, expressed as urea, 2.06%; monoglucosyl urea, 16.7%; combined nitrogen, expressed as monoglucosyl urea, 36%; total nitrogen, 7.0%.
Example VI
This example shows the effect of using a 100%
' excess of urea.
To lOOg of barley hydrolyzate, prepared according to Example I and containing 13g of glucose, were added 8.66g of urea (100% excess) plus 1.73g of 85% phosphoric acid. lOOg ':
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of this mixture were poured into a plastic tray, the thickness of the said mixture being 7 mm, and kept at 75C for 24 hours.
At this stage, 32.9g of solid material were obtained that ~ gave the following analysis: glucose, 0.6~ (conversion of - 5 glucose 98.3~); ammonia plus urea,~expressed as urea, 10.67%
, : monoglucosyl urea, 27.~%; combined nitrogen, expressed as - monoglucosyl urea, 47%; total nitrogen, 12.27%.
Example VII
- This example shows the use of corn as the source of starch in the hydrolysis reaction.
lOOg of corn (whole grain, 18.9% moisture) were mixed with 240 mls of tap water and ground exhaustively.
The pH was adjusted to 4.5 with 85~ phosphoric acid and 30 units of Diazyme L-100 added. Stirring was commenced and the temperature was raised to 67C. At this time, 30 units of Diazyme L-100 were added and the reaction was carried on for 24 hours. A yellow, runny soup was obtained - it contained 14.6% glucose.
Example VIII
This example shows the use of wheat as the source of starch in the hydrolysis reaction.
lOOg of whole ground wheat (12.4% moisture) were mixed with 240 mls of tap water. The pH was adjusted to 4.7 by the addition of 0.6g of 85~ phosphoric acid. 30 units of Diazyme L-100 were added and stirring was commenced.
The temperature was raised to 67C over a period of 1 hour.
The reaction was continued at this temperature and, after i 3 hours, an additional 30 units of Diazyme L-100 were added.
The reaction was then continued for a total reaction time of 16 hours. 340 g of hydrolyzate containing 14% glucose were - obtained.
To lOOg of the wheat hydrolyzate were added : . . . .
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: ~ 10377f~
5.2g of urea (10% excess) plus 1~ of 85% phosphoric acid.
The mixture was placed in a plastic tray to a thickness of 7 mm, and kept at 75C for 14 hours. At this stage, the solid material gave the following analysis: glucose, 4.8%
(conversion 90%); ammonia plus free urea, expressed as urea, 2.39~; ureides, 4 expressed as monoglucosyl urea, 41.63%;
total nitrogen, 8%.
Example IX
This example shows the results of drum-drying.
Barley hydrolyzate containing 13.8~ glucose was rapidly evaporated using a rotary drum under the following conditions:
Thickness of product: 0.025 inches Steam pressure: 48 psi :
Production: 1.45 pounds of dry material per foot 2 per hour ` Slurry feed rate: 4.55 pounds/hour Surface area: 0.962 foot rying time: 37 seconds --In this way, a rapid evaporation of water was ' 20 achieved, however less glucose was converted to ureides - ! compared to the previous examples. A solid product was - obtained that gave the following analysis: glucose, 15%
(conversion 70%); urea plus ammonia, expressed as urea, 5.41%; combined nitrogen, expressed as monoglucosyl urea, 35.0%; monoglucosyl urea, 12.7%; total nitrogen, 8.4%.
! Example X
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This example shows the results of a centrifuge and evaporation trial.
200 g of barley hydrolyzate (13% glucose) plus 10~ excess urea were centrifuged at 200 rpm for 20 hours.
105g of liquid and 105g of cake were obtained. The liquid and the cake were evaporated to yieId 45.15 g of syrup, ,. I
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and 55.?g of cake, respectively; the~ were mixed together to give a mixture with a water content of 35%. This mixture was dried at 75C for 20 hours. 73g of a solid were obtained - that gave the following analysis: glucose, 3% (conversion 90%);
ammonia plus urea, expressed as urea, 3.4%; combined nitrogen, expressed as monoglucosyl urea, 37%, total nitrogen, 8.51%.
Example XI
This example shows the results obtained when the hydrolyzate was cooked instead of dried in accordance with the invention To lOOOg of barley hydrolyzate containing 138g of glucose, were added 50g of urea (10% excess) plus lOg of 85% phosphoric acid. lOOOg of this mixture were evaporated to ; 395g of solid. This solid was placed in a closed container and kept at 75C for 14 hours. At this stage no loss of water was observed. The solid pastry product showed the following analysis: glucose, 8% (conversion of glucose 77%); urea plus ammonia, expressed as urea, 3.98%; combined nitrogen, expressed as monoglucosyl urea, 25%; total nitrogen, 6.76%.
A principal advantage of our product is that it can be produced with a high total equivalent crude protein ` content, with up to 90% of this crude protein in a slow ... .
ammonia release, urease-resistant form. The following examples are from animal feeding trials using various forms of our product and demonstrating the effectiveness of the product.
Example XII
For the purpose of this trial, steers of Here-ford breeding were divided into two groups; one group fed a ration which contained no supplemental protein, the other group fed a ration which contained the solid NPN product prepared as in Example IV. These mixtures constltuted 33%
of the final ration and were formulated to contain similar '~ ..
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levels of all nutrients except crude protein. See Table 1.
- Formulat1on of Diets Ingredients Control Solid non-protein ration nitrogen ration (g~) (%) ~ ,: ~ ' . .. .
Straw (oats) 66.7 66.7 Concentrate 33.3 33.3 Composition of Concentrate Barley 93.7 61.5 - Ground limestone 1.6 2.4 Trace mineralized salt 1.0 1.0 Vitamin premix 11 2.0 2.2 Barley-based non-protein nitrogen -- 32.8 To supply 2,300, 380 and 2.3 IU of Vitamins A, D3 and E, respectively, per pound of total ration.
Compo_ltion of Total Ration Moisture ................. (%) 15.8 15.5 Crude protein............. (%) 5.9 8.9 - Calcium................... (%) 0.3 0.34 Phosphorus................ (~) 0.21 0.26 Digestible energy......... Mcal/pound 1.08 1.05 From Table 2, we see that the steers fed with the solid non-protein nitrogen supplement gained 20% faster than did the animals that received the control ration; they also ate more and needed a lower amount of intake per pound gained.
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T~BLE 2 Effects of Crude Protein Supplementation On the Perfoxmance of Growing Steers - Control Non-protein Nitrogen Number of steers 16 16 Feeding period days 154 154 Av. initial weight pounds 527 530 Av. final weight pounds 612 632 Av. daily gain pounds 0.55 0.66 Av. daily feed pounds 12.5 13.3 Feed/gain 22.47 21.2 . _ _ 12Feed intake is given on an as-fed basis (approximately 84.5%
dry matter).
Example XIII
In this example, three groups of steers were fed rations supplemented with one of the following: (l)our NPN
product, prepared as in example IV; (2)soymeal (which is a premium plant source of protein); and (3)urea. A fourth control~
group was not fed any supplement.
The growing ration used was based on corn silage with a "barley-protein supplemented" concentrate to provide an overall ration with an energy equivalent of approximately 1.17 Mcal per pound of dry matter. The equivalent crude protein content of the unsupplemented control was 0.94 pounds per pound of dry matter, while that of the supplemented rations was 0.11 pounds per pound of dry matter.
The results of the trial, the compositions of the concentrates, and the nutrient content of the feed are given in the following tables:
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1()37768 (Control)(Soymeal)(Test NPN)(Urea) (No supplement) Number of animals 144 143144 144 Days on test 118 105103 119 Pounds Silage consumed per head/day 47.89 50.27 49.30 48.97 Concentrate consumed per head/day 2.99 3.13 3.07 3.06 Pounds total dry matter consumed per head/day14.66 15.39 15.09 14.99 Average total gain (pounds per head)184.7 185.7184.5195.4 Average daily gain1.56 1.77 1.79 1.64 - Conversion ratio (dry matter consumed9.37 8.70 8.43 9.13 per pound gain) ~. . _ . __ _ TABLE IV
Concentrate Composition : .
Low Soybean Test Portein Urea Meal NPN
:........................................ _ ~ Ingredient:
;: 25 Ground Barley 95.35 92.1271.17 74.01 Urea (45% N) -- 3.23 -- --Soybean Meal -- --24.58 --Test NPN -- -- -- 21.56 Dicalcium phosphate -- 0.06 -- ---30 Ground Limestone 2.00 1.941.84 1.95 Plain Salt 1.32 1.321.35 1.86 Beef Mineral/Vitamin Pre. 0.332 0.332 0.332 0.332 Sodium tripolyphosphate 0.710 0.710 0.430 --Elemental Sulfur0.286 0.286 0.286 0.286 - Total 100.0 100.0100.0 100.0 Nutrient:
Dry Matter (%) 88.56 88.9588.78 88.96 ' Crude Protein (%)11.08 19.7719.75 19.78 i Digestible Protein (%) 6.88 -- 15.47 __ ,-40 M.E. Rum. (Mcal/lb)1.144 1.106 1.115 1.101 i TDN (%) 69.56 67.2069.87 66.90 Calcium (%) 0.810 0.797 0.813 0.807 Phosphorus (%) 0.515 0.515 0.519 0.515 .
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lV37~6~3 Sodium (%) 0.749 0.749 0.752 0.761 Magnesium (%) 0.092 0.089 0.135 0.096 Potassium (%~ 0.361 0.349 0.740 0.375 Sulfur (%) 0.481 0.477 0.554 0.481 Iron (mg/lb) 46.9 46.1 56.0 54.4 Copper (mg/lb) 14.5 14.4 15.5 30.5 Manganese (mg/lb)22.4 22.2 24.0 22.7 Zinc (mg/lb) 78.2 77.6 74.3 85.2 Crude Fat (%) 1.8 1.8 1.7 1.4 Crude Fiber (%) 4.8 4.6 4.9 3.7 TABLE V
Nutrient Content of a 1:4- Blend of Concentrate and Corn Silage C.M. (% of DM) Low Soybean Test Protein Urea Meal NPN
----___ _ _ __ Dry Matter (%) 100.0 100.0 100.0 100.0 Crude Protei~ (%)9.43 11.20 11.20 11.20 M.E. (Rum) (Mcal/lb)* 1.176 1.167 1.177 1.166 ;
TDN (%)* 71.55 71.01 71.60 70.95 Calcium (%) 0.379 0.376 0.379 0.377 '-. Phosphorus (%) 0.335 0.334 0.335 0.334 Sodium (%)- 0.211 0.210 0.211 0.213 Magnesium (%) 0.207 0.206 0.175 0.187 Potassium (%) 0.934 0.934 1.01 0.936 Sulfur (%) 0.123 0.122 0.138 0.123 Iron (mg/lb) 115.6 115.40 117.5 117.0 Copper (mg/lb) 5.9 5.9 6.1 9.2 Manganese (mg/lb)20.6 20.5 20.9 20.6 Zinc (mg/lb) 30.5 30.4 29.7 31.9 Fiber (Mg/lb) 22.7 22.7 22.7 22.5 Vitamin A (I.U./lb)3700.03700.03700.0 3700.0 .
; ME - Metabolitable energy TDN - Total digestible nutrients From these data, it will be seen that:
(1) in terms of feed conversion ratio, the . - 19 -, -~- 1~76~
the NPN supplement out-performed all other groups, followed by the soymeal, the urea, ancl the control group. The :: ;
observed difference betw~en the soymeal and the NPN group - and other groups -was statistically significant at the 95%
; confidence level, while the difference between the soymeal and the NPN group was not significant at this level.
(2) the soymeal and the NPN groups gained ~: .
approximately at the same rate, and at a significantly higher rate than the control or the urea group. The urea -; group gained at a significantly faster rate ` 15 than the control group.
(3) in terms of feed intake, the soymeal group consumed significantly more, while the control group consumed significantly ~` less feed than the other groups.
While certain examples~ structures~ composi-tions and process steps have been described for purposes of - illustration the invention is not limited to these. Varia~
tions and modifications within the scope of the disclosure and the claims can readily be effected by those skilled in the art.
SUPPLEM~NTA~Y PISCLOSURE
This supplementary disclosure relates to the use of starch-containing materials other than grains selected from the group consisting of barley, wheat and corn in the process of the Prîncipal Disclosure for producing a non-protein nitrogen-containing material useful as a feed supplement for cattle and sheep. The starch-containing materials may ~5 ~37~6~
i include different grains, such as rice and oats, as well as sources other than grains, such as cassava roots and potatoes.
; The characteristics of various suitable starch sources are given in Table 7.
TAsLE 7 Starch ~oisture Starch Source (%) - (%, d.m.b.) barley 10.8 59.9 sorghum 8.5 57.9 10 tapioca 10.0 96.6 rice 11.9 95.6 oats 10.5 53.3 : corn 9.8 82.1 cassava root 7.6 93.7 15 potatoes 78.4 69.5 The invention is not restricted to materials which hydrolyze to produce glucose as the major hydrolysis product. Glycoses, HOCH2 ~CHO~)nCHO, other than glucose can be ~ -reacted with urea to produce glycosyl ureides. For example, the spent sulphite liquours of the paper industry containxylose in amounts greater than glucose and can be used in the process of the present invention.
Broadly stated, the invention is a process for producing a non-protein nitrogen feed supplement for ruminants which comprises: providing an aqueous mixture of a starting - material containing one or more reducing carbohydrates as the major dry matter constituent, and urea; and reacting the mixture under acidic and drying conditions at an elevated temperature sufficient to convert the major portion of the reducing carbohydrates and urea into ureides and to produce a solid product.
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ANALYS I S` PROCEDURES
, Analysis procedures similar to those described in the Principal Disclosure were used unless otherwise specified.
The concentration of total ureides was deter-mined by acid hydrolysis of the product to give glucose and urea, followed by enzymatic hydrolysis of the urea and by the ~- potentiometric measurement of the resulting ammonia. The -`, measurement gave the total ammonia concentration; this value was corrected for the amount of free ammonia and free urea to ~,. 1 ~X` 10 obtain the total ureide concentration.
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The moisture content of a starch source was determined by heating a sample in an oven at 75C to constant weight.
Starch determination was done by treatment of a known amount of the starcll source suspended in water with ; 85% phosphoric acid (pH 1.5 - 2.0) at 80C overnight. A
control experlment was simultaneously conducted using pure potato staxch. After cooling and adjusting the p~I to 4.5 with sodium hydroxide, the unknown and the control samples were treated with the same amount of glucoamylase at 62C for 24 hours. The glucose thus produced was analyzed, and the result obtained was corrected according to the glucose obtained from the control experlment.
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THE HYDROLYSIS REACTION
The hydrolysis reaction was carried out in equipment similar to that described in the Principal Disclosure.
The standard reaction conditions were~
100 g substrate, crushed to 2/64"
24Q ml distilled water pH 4.0 to 4.5 (by use of 85% phosphoric acid) 1.0 ml Diazyme L-100* added in two batches;
0.5 ml at zero time and 0.5 ml three hours after reaction temperature reached 65C
reaction time: 24 hours The relatively low conversions to glucose in the case of sorghum and corn prompted the pre-treatment of the substrate at high temperature and acidic conditions overnight, prior to hydrolysis. In all cases, this procedure substantially increased the conversion of starch to glucose.
It was also found that the mixing of the sub-strate with water in a ~aring* Blender prior to the hydrolysis reaction also increased thé conversion to glucose. This was tried in the case of sorghum, corn, and potatoes.
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- Due to the high moisture content, potatoes were the only starch source which was not crushed prior to reaction.
Instead, the potatoes were cut into small pieces with a knife and mixed with water in a Waring* slender. The soup obtained after hydrolysis was extremely runny, indicating that far less than the standard 240 mls of water could be used. A
reaction using only 120 mls of water was tried; this also pro-duced a very runny soup.
- The standard reaction time of 24 hours was cut to 6 hours when it was discovered that ~he heater malfunctioned overnight. During the 6-hour reaction it was observed that the controller maintained the temperature constant to within - 1.0C. The average increase in glucose content between 6 hours of reaction and 24 hours of reaction was 2.2~ based on analysis of the first 13 hydrolyses in which the heater was thought to have Punctioned properly.
THE CONDENSATION REACTION
The condensation reaction was carried out by mixing the hydrolyzate with a lQ~ stoichiometric excess of urea and with phosphoric acid ta obtain a pH of 2.5 - 3Ø The mixture was spread on a plastic tray and heated at a given temperature to dryness. The product was analyzed for glucose and monoglucosyl ureides.
The following examples illustrate the use of various starch-containing materials in the process of the present lnvention.
Example 14 This example shows the use of sorghum as the source of starch in the hydrolysis reaction.
: 30 10-Q g of crushed sorghum were mixed with 240 mls of distilled water. The pH was adjusted to 2.0 with 85~ -~ .
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phosphoric acid and the mixture was left at 80C overnight.
After cooling and adjusting the pH to 4.5 with sodium hydroxide, :.
50 units of Diazyme L-100* were added. Stirring was commenced and the temperature was raised to 65C. Three hours a~ter having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 15%.
5.5 g of urea (a 10% excess) and 2 g of 85%
; 10 phosphoric acid were added to 100 g of the sorghum hydrolyzate.
The mixture was spread in a plastic tray to a thickness of 7 mm, ~ ~ -and kept at 75C for 2a hours. At this stage~ the solid -material gave the following analysis: glucose, 3% (93% con~
version); ammonia, expressed as urea, 0.44%; ammonia plus free -urea, expressed as urea, 5.6%; ureides, expressed as mono-glucosyl urea, 34.8% com~ined nitrogen, expressed as mono-glucosyl urea, 42~1%; total nitrogen 8.51%.
Example 15 -; This example shows the use of oats as the source of starch in the hydrolysis reaction.
100 g o~ crushed oats were mixed with 24Q mls ofdistilled water. The pH was adjusted to 4.5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 5Q units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the h~drol~zate was found to be 15.g%.
5.84 g of urea (a 10% excess) and 2 g of 85%
phosphoric ac~d were added to 100 g of the oat hydrolyzate.
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7~ 3 The mixture was spread in a plastic tray to a thickness of 7mm, and kept at 75C for 70 hours. At this stage, the solid material gave the following analysis: glucose, 1~ (98% con-version); ammonia, expressed as urea, 0.58%; ammonia plus free urea, expressed as urea, 5.0%; ureides, expressed as mono-glucosyl urea, 38.9%; combined nitrogen, expressed as mono-glucosyl urea, 45.0%; total nitrogen, 9.52%.
Example 16 This example shows the use of rice as the source of starch in the hydrolysis reaction.
lQ0 g of crushed brown rice were mixed with 240 mls of distilled water. The pH was adjusted to 4~5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
Stirring was commenced and the temperature was raised to 65C.
Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 21 4%
- 20 7~84 g o~ urea (lQ% excess) and 3 g of 85% -phosphoric acid were added to 100 g of the rice hydrolyzate.
The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 75~C for 50 hours. At thîs stage, the solid material gave the following analysis: glucose, 1% (98% con-version~; ammonia, expressed as urea, 0.78%; ammonia plus free urea, expressed as urea, 5.5%; ureides, expressed as mono-; glucosyl urea, 37.0%; combined nitrogen, expressed as mono-;~ glucosyl urea, 61.5%; total nitrogen, 11.3%.
Example 17 . '.
` 30 This example shows the use of corn as the source of starch in the hydrolysis reaction.
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lQQ g of crushed corn were mixed with 240 mls ~ -of distilled water and ground exhaustively. The pH was ad-justed to 4.5 with 85% phosphoric acid and 50 units of Diazyme L-100* were added. Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 6 hours, at which stage the glucose content of the hydrolyzate was found to be 10.8%.
4.34 ~ of urea (10% excess) and 1 g of 85%
phosphoric acid were added to 100 g of the corn hydrolyzate~
The mixture ~as spread in a plastic tray to a thickness of 7 mm, and kept at 75C for 22 hours. At this stage, the solid material gave the following analysis: glucose, 2~4% (94%
conversion); ammonia, expressed as urea, 0.15%; ammonia plus -free urea, expressed as urea, 3.0%; ureides, expressed as monoglucosyl urea,-25.9%; combined nitrogen, expressed as monoglucosyl urea, 44.5%; total nitrogen, 8.35%.
The results of the hydrolysis of various grains and the analyses of the corresponding condensation products are given in Table 8.
Glucosyl Glucose Conver- ureide in hydro- sion to (GU) in Yield Glucose lyzate glucose product of GU reacted Substrate ` (~O) (%) (~O~ (%) (%) barley 17.8 100 14-3 } 16 hrs.
16.3 at 75C
14.5 93.2 19-6 l 24 hrs. 85.1 18.3 J at 60 32.3 84.9 -1: .
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1~)37~7f~3 TABLE 8 (continued) Glucosvl ` Glucose Conver- ureide ' in hydro- sion to (GU) in Yield Glucose lyzate glucose product - of GU reacted Substrate(~ ) (%) - (~-) ` (~) sorghum 3.0 18.03.3 90 hrs. at 23.1 78.4 15.0a 93.216.3l 20 hrs. 28.8 93.5 12.5J at 75 22.4 68.1 b 9.6 71.1 4.9 24 hrs. 12.8 95.2 at 60 3.5 19.2 3.4 50 hrs. 24.0 65.1 at 65 4.7c,d 25.8 6.7 16 hrs. 34.5 79.0 at 75 4.2 16 hrs. 21.5 53.6 at 65 .: ' .
tapioca18~2 65.0 25.0 68 hrs 38.6 95.1 at 60 22.2 17 hrs. 34.1 92.8 ; at 60-80 ; 25 15.6 138 hrs. 23.9 97.4 at 60 20.8 51 hrs. 31.9 97.5 at 65 ;;
rice 11.9 43.1 7.2 138 hrs. 15.0 93.8 - at 60 19.6 27 hrs. 42.4 94.1 - at 65 J
18.6a 72.9 <4 5 days ~6 97.9 at 65 21.4 73.8 19.8 50 hrs. 26.8 98.2 ' at 75 `
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22.5 75 hrs. 31.8 97.9 at 65 ~ ~',' '.
Example 18 ;~
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This example shows the use of a starch-con-taining material that is not a grain.
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~76 - 100 g of crushed cassava root were mixed with 240mls of distilled water. The pH was adjusted to 4.5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
- Stirring was commenced and the temperature was raised to 65C.
Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 21.1%.
7.74 g of urea (a 10% excess) was added to 100 g of the cassava root hydrolyzate, and the pH was adjusted to 2.5 with 85% phosphoric acid. The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 60C for 120 hours.
~ At this stage, the solid material gave the following analysis:
.~ glucose, 1% (98% conversion~; ar,~onia, expressed as urea, 0.33%;
ammonia plus free urea, expressed as urea, 5.4%; ureides, ex-pressed as monoglucosyl urea, 41.4%; combined nitrogen, expressed as monoglucosyl urea, 57.~P6; total nitrogen, 9.27%.
Example 19 This example shows the use of starch-containing - 2~ material that is not a grain.
100 g of diced potatoes were mixed with 240 mls of distilled water an~ ground exhaustively. The pH was adjusted to 4.5 with 85% phosphoric acid and 50 units of Diazyme L-100*
~; were added. Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added.
The reaction was then continued for a total reaction time of 6 hours, at which stage the glucose content of the hydrolyzate was found to be 3.7%.
1.36 g of urea (a 100% excess) and 1 g of 85%
phosphoric acid were added to 100 g of the potato hydrolyzate.
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~ * trademark - 28 -;' The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 75C for 18 hours. At this stage, the solid material gave the following analysis: glucose, 1% (98% conversion);
ammonia, expressed as urea, 0.29%; ammonia plus free urea, expressed as urea, 1.7%; ureides, expressed as monoglucosyl urea, 30.7%; combined nitrogen, expressed as monoglucosyl urea, 52.1%; total nitrogen, 8.26%.
Exà~ple 20 .
This example shows the use of spent sulphite liquor as a source of reducing sugars in the condensation reaction.
The spent liquor used was typical of that obtained from a hard-wood pulp, and contained approximately 26% xylose on a dry matter basis.
25 g of spent sulphite liquor (40% moisture) having a pH of 2.0 were mixed at room temperature with 1.8 g urea. The mixture was spread on a plastic tray to a thickness of 7 - 8 ~m, and kept at 75C for 110 hours. At this stage, the product gave the followlng analysis: free ammonia plus free - urea, expressed as urea, 5.2%; ureides, expressed as equivalent ~ 20 monoglucosyl urea, 17.8%; total nitrogen, expressed as urea, -. 10. 0% .
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phosphoric acid were added to 100 g of the corn hydrolyzate~
The mixture ~as spread in a plastic tray to a thickness of 7 mm, and kept at 75C for 22 hours. At this stage, the solid material gave the following analysis: glucose, 2~4% (94%
conversion); ammonia, expressed as urea, 0.15%; ammonia plus -free urea, expressed as urea, 3.0%; ureides, expressed as monoglucosyl urea,-25.9%; combined nitrogen, expressed as monoglucosyl urea, 44.5%; total nitrogen, 8.35%.
The results of the hydrolysis of various grains and the analyses of the corresponding condensation products are given in Table 8.
Glucosyl Glucose Conver- ureide in hydro- sion to (GU) in Yield Glucose lyzate glucose product of GU reacted Substrate ` (~O) (%) (~O~ (%) (%) barley 17.8 100 14-3 } 16 hrs.
16.3 at 75C
14.5 93.2 19-6 l 24 hrs. 85.1 18.3 J at 60 32.3 84.9 -1: .
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1~)37~7f~3 TABLE 8 (continued) Glucosvl ` Glucose Conver- ureide ' in hydro- sion to (GU) in Yield Glucose lyzate glucose product - of GU reacted Substrate(~ ) (%) - (~-) ` (~) sorghum 3.0 18.03.3 90 hrs. at 23.1 78.4 15.0a 93.216.3l 20 hrs. 28.8 93.5 12.5J at 75 22.4 68.1 b 9.6 71.1 4.9 24 hrs. 12.8 95.2 at 60 3.5 19.2 3.4 50 hrs. 24.0 65.1 at 65 4.7c,d 25.8 6.7 16 hrs. 34.5 79.0 at 75 4.2 16 hrs. 21.5 53.6 at 65 .: ' .
tapioca18~2 65.0 25.0 68 hrs 38.6 95.1 at 60 22.2 17 hrs. 34.1 92.8 ; at 60-80 ; 25 15.6 138 hrs. 23.9 97.4 at 60 20.8 51 hrs. 31.9 97.5 at 65 ;;
rice 11.9 43.1 7.2 138 hrs. 15.0 93.8 - at 60 19.6 27 hrs. 42.4 94.1 - at 65 J
18.6a 72.9 <4 5 days ~6 97.9 at 65 21.4 73.8 19.8 50 hrs. 26.8 98.2 ' at 75 `
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Example 18 ;~
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This example shows the use of a starch-con-taining material that is not a grain.
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~76 - 100 g of crushed cassava root were mixed with 240mls of distilled water. The pH was adjusted to 4.5 with 85%
phosphoric acid and 50 units of Diazyme L-100* were added.
- Stirring was commenced and the temperature was raised to 65C.
Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added. The reaction was then continued for a total reaction time of 24 hours, at which stage the glucose content of the hydrolyzate was found to be 21.1%.
7.74 g of urea (a 10% excess) was added to 100 g of the cassava root hydrolyzate, and the pH was adjusted to 2.5 with 85% phosphoric acid. The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 60C for 120 hours.
~ At this stage, the solid material gave the following analysis:
.~ glucose, 1% (98% conversion~; ar,~onia, expressed as urea, 0.33%;
ammonia plus free urea, expressed as urea, 5.4%; ureides, ex-pressed as monoglucosyl urea, 41.4%; combined nitrogen, expressed as monoglucosyl urea, 57.~P6; total nitrogen, 9.27%.
Example 19 This example shows the use of starch-containing - 2~ material that is not a grain.
100 g of diced potatoes were mixed with 240 mls of distilled water an~ ground exhaustively. The pH was adjusted to 4.5 with 85% phosphoric acid and 50 units of Diazyme L-100*
~; were added. Stirring was commenced and the temperature was raised to 65C. Three hours after having reached this temperature, an additional 50 units of Diazyme L-100* were added.
The reaction was then continued for a total reaction time of 6 hours, at which stage the glucose content of the hydrolyzate was found to be 3.7%.
1.36 g of urea (a 100% excess) and 1 g of 85%
phosphoric acid were added to 100 g of the potato hydrolyzate.
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~ * trademark - 28 -;' The mixture was spread in a plastic tray to a thickness of 7 mm, and kept at 75C for 18 hours. At this stage, the solid material gave the following analysis: glucose, 1% (98% conversion);
ammonia, expressed as urea, 0.29%; ammonia plus free urea, expressed as urea, 1.7%; ureides, expressed as monoglucosyl urea, 30.7%; combined nitrogen, expressed as monoglucosyl urea, 52.1%; total nitrogen, 8.26%.
Exà~ple 20 .
This example shows the use of spent sulphite liquor as a source of reducing sugars in the condensation reaction.
The spent liquor used was typical of that obtained from a hard-wood pulp, and contained approximately 26% xylose on a dry matter basis.
25 g of spent sulphite liquor (40% moisture) having a pH of 2.0 were mixed at room temperature with 1.8 g urea. The mixture was spread on a plastic tray to a thickness of 7 - 8 ~m, and kept at 75C for 110 hours. At this stage, the product gave the followlng analysis: free ammonia plus free - urea, expressed as urea, 5.2%; ureides, expressed as equivalent ~ 20 monoglucosyl urea, 17.8%; total nitrogen, expressed as urea, -. 10. 0% .
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Claims (6)
1. A process for producing a non-protein nitrogen feed supplement for ruminants which comprises:
providing an aqueous mixture of a starting material containing one or more reducing carbohydrates as the major dry matter constituent, the major constituent of said reducing carbohydrates being glucose, and urea; and reacting the mixture under acidic and drying conditions at an elevated temperature sufficient to convert the major portion of the glucose and urea into glucosyl ureides and to produce a solid product.
providing an aqueous mixture of a starting material containing one or more reducing carbohydrates as the major dry matter constituent, the major constituent of said reducing carbohydrates being glucose, and urea; and reacting the mixture under acidic and drying conditions at an elevated temperature sufficient to convert the major portion of the glucose and urea into glucosyl ureides and to produce a solid product.
2. A process for producing a non-protein nitrogen feed supplement for ruminants which comprises:
mixing glucose, produced by the hydrolysis of starch in grain selected from the group consisting of barley, wheat and corn, said glucose still being associated with the other normal constituents of the grain, with water, urea and sufficient mineral acid to provide a mixture having a starting pH of about 2 - 4, said glucose being the major dry matter constituent of said mixture; and drying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the glucose to react and form urease - resistant glucosyl ureides in solid form.
mixing glucose, produced by the hydrolysis of starch in grain selected from the group consisting of barley, wheat and corn, said glucose still being associated with the other normal constituents of the grain, with water, urea and sufficient mineral acid to provide a mixture having a starting pH of about 2 - 4, said glucose being the major dry matter constituent of said mixture; and drying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the glucose to react and form urease - resistant glucosyl ureides in solid form.
3. A feed supplement for ruminants comprising a solid, condensation reaction product formed by the reaction of claim 1, said product containing the major proportion of said glucose in the form of glucosyl ureides.
Claims Supported by Supplyementary Disclosure
Claims Supported by Supplyementary Disclosure
4. A process for producing a non-protein nitrogen feed supplement for ruminants which comprises:
providing an aqueous mixture of a starting material containing one or more reducing carbohydrates, as the major dry matter constituent, and urea; and reacting the mixture under acidic and drying con-ditions at an elevated temperature sufficient to convert the major portton of the reducing carbohydrates and urea into ureides and to produce a solid product.
providing an aqueous mixture of a starting material containing one or more reducing carbohydrates, as the major dry matter constituent, and urea; and reacting the mixture under acidic and drying con-ditions at an elevated temperature sufficient to convert the major portton of the reducing carbohydrates and urea into ureides and to produce a solid product.
5. A process for producing a non-protein nitro-gen feed supplement for ruminants which comprises:
mixing glucose, produced from the hydrolysis of starch in one or more materials selected from the group consist-ing of barley, wheat, corn, sorghum, tapioca, cassava roots, rice, oats and potatoes, said glucose still being associated with the other normal constituents of the material, with water, urea and sufficient mineral acid to provide a mixture having a starting pH of about 2 - 4, said glucose being the major dry matter constituent of said mixture; and drying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the glu-cose to react and form urease-resistant glucosyl ureides in solid form.
mixing glucose, produced from the hydrolysis of starch in one or more materials selected from the group consist-ing of barley, wheat, corn, sorghum, tapioca, cassava roots, rice, oats and potatoes, said glucose still being associated with the other normal constituents of the material, with water, urea and sufficient mineral acid to provide a mixture having a starting pH of about 2 - 4, said glucose being the major dry matter constituent of said mixture; and drying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the glu-cose to react and form urease-resistant glucosyl ureides in solid form.
6. A process for producing a non-protein nitrogen feed supplement for ruminants which comprises:
providing an aqueous-mixture of xylose-containing material and urea at a starting pH of about 2 - 4, said xylose being the major dry matter constituent of said mixture; and dxying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the xylose to react and form urease-resistant xylosyl ureides in solid form.
providing an aqueous-mixture of xylose-containing material and urea at a starting pH of about 2 - 4, said xylose being the major dry matter constituent of said mixture; and dxying the mixture at an elevated temperature for sufficient time to cause urea and the major portion of the xylose to react and form urease-resistant xylosyl ureides in solid form.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA216,798A CA1037768A (en) | 1974-12-24 | 1974-12-24 | Ruminant feed supplement and process for producing same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA216,798A CA1037768A (en) | 1974-12-24 | 1974-12-24 | Ruminant feed supplement and process for producing same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1037768A true CA1037768A (en) | 1978-09-05 |
Family
ID=4101929
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA216,798A Expired CA1037768A (en) | 1974-12-24 | 1974-12-24 | Ruminant feed supplement and process for producing same |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA1037768A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2556566A1 (en) * | 1983-12-15 | 1985-06-21 | Corning Glass Works | NEW PROCESS FOR PREPARING A GLYCOSYL-UREA FOOD SUPPLEMENT FOR RUMINANTS AND SUPPLEMENT FOOD OBTAINED |
| US5233997A (en) * | 1991-04-04 | 1993-08-10 | Baylor College Of Medicine | Non-invasive measure of intestinal transit time and uses thereof |
-
1974
- 1974-12-24 CA CA216,798A patent/CA1037768A/en not_active Expired
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2556566A1 (en) * | 1983-12-15 | 1985-06-21 | Corning Glass Works | NEW PROCESS FOR PREPARING A GLYCOSYL-UREA FOOD SUPPLEMENT FOR RUMINANTS AND SUPPLEMENT FOOD OBTAINED |
| US5233997A (en) * | 1991-04-04 | 1993-08-10 | Baylor College Of Medicine | Non-invasive measure of intestinal transit time and uses thereof |
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