CN102578394A - Application of cupric glutamate or derivative of cupric glutamate serving as feed additive for promoting animal growth - Google Patents
Application of cupric glutamate or derivative of cupric glutamate serving as feed additive for promoting animal growth Download PDFInfo
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- CN102578394A CN102578394A CN201210075538XA CN201210075538A CN102578394A CN 102578394 A CN102578394 A CN 102578394A CN 201210075538X A CN201210075538X A CN 201210075538XA CN 201210075538 A CN201210075538 A CN 201210075538A CN 102578394 A CN102578394 A CN 102578394A
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Abstract
The invention discloses application of cupric glutamate or a derivative of the cupric glutamate serving as a feed additive for promoting animal growth and finds that the cupric glutamate has low water-solubility, strong acid-resisting capability and high bioavailability. Compared with other copper resources, such as copper sulfate, basic copper chloride and glycine, the cupric glutamate serving as the feed additive for promoting animal growth has higher safety and antibacterial activity, the copper ion adding amount can be reduced by 60-80%, production cost and environmental pollution can be reduced, and simultaneously toxic and side effects of high copper to animals are reduced.
Description
Patent application of the present invention is that application number is dividing an application of " 201010122625.7 "; The applying date of original application is " on 03 12nd, 2010 "; Application number is " 201010122625.7 ", and denomination of invention is " the cupric glutamate or derivatives thereof is as the application of animal feed additive for promoting growth ".
Technical field
The present invention be more particularly directed to of the application of a kind of cupric glutamate or derivatives thereof, belong to field of fodder as animal feed additive for promoting growth.
Background technology
Copper is one of essential trace element of animal, participates in hematopoiesis, metabolism, growth and breeding and the activity such as disease-resistant of animal.Animal to the nutritional requirements of copper at 5-8mg/kg; But since 1945 find that high-copper can improve the growth performance of pig significantly; All adopt the high-copper (in copper ion) of 200-250ppm (quality) at present in children's piglet stage in age, high-copper has the effect of the secretion of solidifying protein, the propagation that reduces microorganism, the peptide Y (NPY) that excites nerve, thereby promotes that animal searches for food; Improve digestive enzyme activity, finally improve the growth performance effect.So pig has become many nutritionists " custom " so far in the 1950's with high-copper.
At present; The source master of pig starter feed high-copper is inorganic copper sources such as copper sulphate, basic copper chloride or basic copper sulfate; Inorganic copper source itself water-soluble or its can be decomposed by hydrochloric acid in gastric juice in the pig stomach fast; Thereby a large amount of free copper copper ions discharges fast under one's belt and is absorbed, thereby the amount of giving birth to that surpasses animal is limited the quantity of and caused toxic reaction.Yet in absorbed a large amount of free copper ion, have only a spot of copper ion can arrive the small intestine postmedian and play antimicrobial effect, thereby the copper ion that need in feed, add high concentration just can play antibiotic somatotrophic effect.But, use high-copper to bring many side effects, mainly show as: (1) high-copper is to the infringement of body internal organs; Use the 200ppm high-copper can cause the liver cell enlargement, cytoplasmic granule sex change, glomerulus enlargement; Renal tubule swelling, capsula glomeruli oozes out albumen, the cardiac muscle fibre granular degeneration; Lymphocyte reduces under the spleen, the smooth muscle fibers hyperplasia, through evidence the high-copper weanling pig that the low copper of the comprehensive sick ratio of dermatitis ephrosis takes place is high more than 5%; Explaining that high-copper is one of inducement of the scorching nephrotic syndrome of pigskin, also is to cause one of reason that the pig resistance descends; (2) high-copper residual exceeding standard in organ influences the security of food.The copper sanitary standard (GB15199-94) of limiting the quantity of is no more than 10mg/kg in China's meat product, but in the piglet basal diet, adds the 250ppm high-copper, and copper content is 186.27mg/kg in the liver; (4) absorption of other nutrition of high-copper interfere causes pig hypovitaminosis like vitamin and grease in the high-copper ability oxidation feed, and thick unrest of fur and feed quality descend; High-copper antagonism iron, zinc absorb and cause hypoferric anemia and pachylosis, zinc deficiency symptom such as are taken off by hair; (4) high-copper also has huge environmental pressure, and in ten thousand pig farms, annual compound of discharging copper reaches 2.5 tons, and high-copper poultry pollution underground water source makes fresh-water fishes dead, and melon and fruit, vegetable crop is sterile or yield poorly.
Summary of the invention
The objective of the invention is to overcome that employed inorganic copper source provides the application of a kind of cupric glutamate or derivatives thereof as animal feed additive for promoting growth as the weak point of animal feed additive for promoting growth in the prior art.
The object of the invention is realized through following technical proposals: a kind of cupric glutamate or derivatives thereof is as the application of animal feed additive for promoting growth.
Described cupric glutamate comprises: glutamic acid and copper 2: 1 in molar ratio or the chelate that forms at 1: 1.
Described derivative comprises:
(1) cupric glutamate and the different sour different salt that form are like the hydrochloride of cupric glutamate, the phosphate of cupric glutamate and the sulfate of cupric glutamate etc.;
(2) glutamic acid becomes inner complex salt or the chelate that forms with copper ion behind the monoesters;
(3) chelate or the salt of N-carbamylglutamic acid and different salt, acid amides and copper ion formation;
(4) with the cupric glutamate be the different shiff alkali that the basis forms;
(5) with the cupric glutamate be the different amide compounds that the basis forms;
(6) with the dipeptides of a kind of formation in glutamic acid and the amino acid and the chelate of copper ion formation;
(7) chelate or the salt of the formation of glutamine and copper ion.
Described animal comprises various cultivated animals, as: various artificial feeding animals such as pig, chicken, duck, goose, beef cattle, milk cow, sheep, various fish and shrimp, fox, ermine, racoon dog.
Said cupric glutamate or derivatives thereof is applied to the different growth phases of animal as the application of animal feed additive for promoting growth.
When described animal feed is perfect compound feed, add cupric glutamate as growth promoter, the consumption of cupric glutamate is in copper, and its content is 20～100ppm.
The present invention has following advantage and effect with respect to prior art: the present invention finds the effect of cupric glutamate or derivatives thereof as animal feed additive for promoting growth first.The present invention finds that cupric glutamate has low aqueous solubility, stronger acidproof ability and high bioavilability; With cupric glutamate as feed additive for promoting growth; Have higher security and antibacterial activity than other copper source (comprising copper sulphate, basic copper chloride and cupric glycinate etc.); The addition of copper ion can reduce 60～80%, can reduce production costs and reduce pollution to environment, reduce the toxic and side effect of high-copper to animal simultaneously.
The specific embodiment
Below in conjunction with embodiment the present invention is described in further detail, but embodiment of the present invention is not limited thereto.
Embodiment 1
(1) synthetic cupric glutamate:
The preparation of A, 10% monosodium glutamate solution: take by weighing 100 gram sodium glutamates (chemical pure) and be dissolved in the 1000ml water;
The preparation of B, 13.37% copper-bath: accurately smart amount 133.7 gram cupric sulfate pentahydrates (chemical pure) are dissolved in the 1000ml water;
C, each 1000ml of 10% monosodium glutamate solution and 13.37% copper-bath is mixed, the pH value to 7.0 of then regulating aforementioned mixed solution with NaOH solution, stirring at room is reacted 2h; Centrifugal; Get deposition, water suspended centrifugal washing precipitation 3 times is then dried to constant weight for 100 ℃; Obtain the 117.4g cupric glutamate, yield is 96.31%.
The glutamic acid of the cupric glutamate that (2) step (1) is obtained and the content of copper ion are measured respectively:
The mensuration of A, content of copper ion: accurately take by weighing the 1.0g cupric glutamate; Add water 80ml, use 0.1M HCl adjust pH to 2.0 then, be stirred to cupric glutamate and dissolve fully; With the content of copper ion in the atom absorption method mensuration solution, recording copper ion concentration is 2.807mg/ml.
The assay of B, glutamic acid: accurately take by weighing 1.0 gram cupric glutamates; Add water 80ml, use 0.1MHCl adjust pH to 2.0 then, be stirred to cupric glutamate all after the dissolving; With the content of amino-acid analyzer mensuration glutamic acid, the content that records glutamic acid is 6.404mg/ml.
Thereby the mol ratio of copper ion and glutamic acid is 1: 1 in definite cupric glutamate, and crystallization water content is 7.89% in the cupric glutamate, and therefore, the molecular formula of the cupric glutamate that step (1) obtains is C
5H
8NO
4CuH
20, molecular weight is 228.
(3) the antibacterial activity in vitro research of the cupric glutamate that obtains of step (1)
1. test material
A, culture medium: LB fluid nutrient medium: the 10g tryptone, 5g yeast extract and 10g NaCl are dissolved in the 800ml distilled water, with being settled to 1000mL behind the 1M NaOH adjust pH to 7.4, autoclaving 20min;
B, bacterial strain: e. coli jm109, salmonella typhimurium 50772 and staphylococcus aureus PNB14, all preserve the center available from China Veterinary Drugs Supervisory Inst.'s bacterial classification;
C, test tube: 10ml teat glass with cover;
D, medicine: CuSO
45H
2O solution: 0.2% (in copper ion, the mass/volume percent concentration, down together); Basic copper chloride (Cu
2Cl. (OH)
3): 0.2% (, using the 0.01M dissolving with hydrochloric acid) in copper ion; Cupric glutamate solution: 0.2% (with the DMSO dissolving, in copper ion);
2. test method: determination of tube method copper sulphate, basic copper chloride and cupric glutamate are to the antibacterial activity of e. coli jm109, salmonella typhimurium 50772 and staphylococcus aureus PNB14:
A, establish 9 groups, every group comprises 12 sterile test tube, numbering 1～12; Every big group establish three parallel, promptly comprise three groups;
Under B, the aseptic condition, add 2.0ml LB fluid nutrient medium to the 1 to 11 pipe respectively;
C, the copper sulphate for preparing, basic copper chloride or cupric glutamate solution to be checked is added 2.0ml to the 1 pipe respectively; With getting 2.0ml to the 3 pipes after the 1st pipe mixing; To the 10th pipe, get 2.0 milliliters from the 10th pipe again and lose successively, the 11st pipe is made positive control for not adding antiradiation drug;
C, prepare LB Liquid Culture parent tube (the 12nd pipe) 2.0ml in addition and do not add medicine and bacterium, as negative control;
The 1-11 pipe of D, every big group adds e. coli jm109, salmonella typhimurium 50772 and the staphylococcus aureus PNB14 that supplies test respectively, and (concentration of bacterium is about 10 respectively to every pipe 5.0 microlitre bacterium liquid
8Cfu/ml, cell age is 16-18 hour);
E, 37 ℃ of static cultivations 16 hours, perusal has or not bacterial growth, and last pipe Chinese traditional medicine concentration that bacterial growth do not occur promptly is the minimal inhibitory concentration (μ g/ml) of medicine to corresponding bacterium; Positive control is answered the visible haze growth, and negative control should be clarified.
3. result of the test:
In copper ion; Copper sulphate and basic copper chloride are 500 μ g/ml, 500 μ g/ml and 500 μ g/ml (in copper ion) to the minimal inhibitory concentration of e. coli jm109, salmonella typhimurium 50772 and staphylococcus aureus PNB14; And cupric glutamate to the minimal inhibitory concentration of e. coli jm109, salmonella typhimurium 50772 and staphylococcus aureus PNB14 for being respectively 125 μ g/ml, 125 μ g/ml and 125 μ g/ml (in copper ion), antibacterial activity has improved 4 times.
The antibacterial activity comparative studies result of table 1 copper sulphate and cupric glutamate
(4) (rat LD is measured in the security of the cupric glutamate that obtains of step (1)
50Measure, improve karber's method)
1. test material
Rat: available from Nanfang Medical Univ's Experimental Animal Center, body weight 120～150 grams, male and female half and half.
Supply test specimen: cupric sulfate pentahydrate, basic copper chloride and cupric glutamate
Equipment: syringe, filling stomach syringe needle, mouse cage
2. test method
A, obtain causing animal 0% (Dn) and the dead dosage of 100% (Dm) through preliminary experiment, wherein the Dn of cupric sulfate pentahydrate, basic copper chloride and cupric glutamate and Dm see table 2.
Table 2 different Cu source compound rat ID50 trial test result (Dn and Dm)
| Compound | Dn (mg/kg body weight) | Dm (mg/kg body weight) |
| Cupric sulfate pentahydrate | 1000 | 1800 |
| Basic copper chloride | 1200 | 2000 |
| Cupric glutamate | 10000 | 15000 |
B, this requirement of experiment maximum response rate are 100%, and the minimal reaction rate is 0%, or at least reactivity near 100% or 0%; Dose ratio is used Dn and the Dm value according to above-mentioned each compound between group, and 8 dosage on average are set between Dn to Dm, every group of 10 rats.When 100% and 0% reactivity of repetition being arranged, should the group of keeping to the side be discarded and disregard as adjacent doses occurring in the experiment, the reactivity that makes heavy dose of group have only 100%, small dose group also has only one 0% reactivity; After dividing into groups to finish, divide into groups to irritate copper sulphate, basic copper chloride or the cupric glutamate of clothes various dose; Observed 7 days after the administration, viewing duration writes down the toxic reaction situation of animal and the distribution of dead animal day by day
C, account form are respectively organized the death rate by LD50 and the fiducial limit (P=0.95) of following formula different Cu source compound to rat according to formal experiment
A, when the death rate of minimum dose group is 0%, the death rate of maximum dose group is 100% o'clock, calculates LD50 by following formula:
LD
50＝lg
-1[Xm-i(∑p-0.5)]
B, when the death rate of minimum dose group greater than 0% and less than 30%, or the death rate of maximum dose group can be calculated LD by following updating formula less than 100% and greater than 70% o'clock
50:
LD
50Standard error:
LD
50Average fiducial limit: LD
50± 4.5S
X50LD
50(P=0.95);
In the following formula, Xm is the logarithm of maximum dose group dosage, and i is the difference (heavy dose of group reduces dose groups) of two adjacent groups log10 dose, and Pm is the maximum dose group death rate, and Pn is the minimum dose group death rate, and P is each group death rate, and n is every treated animal number.
D, result of the test
In copper ion, rat sees that to the dead dosage of oral half (LD50) of all given the test agent table 3, result show that cupric glutamate improves respectively about 10 times and 4 times than copper sulphate and the right security of basic copper sulfate.
Table 3 copper sulphate, basic copper chloride and cupric glutamate are to the oral median lethal dose result of study of rat
| Given the test agent | LD50 (the mg/kg body weight is in compound) | LD50 (the mg/kg body weight is in copper ion) |
| Cupric sulfate pentahydrate | 1300 | 332.8 |
| Basic copper chloride | 1500 | 900 |
| Cupric glutamate | 12000 | 3383.3 |
(5) mensuration of the cupric glutamate equilbrium solubility that obtains of step (1)
1. test material
Potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate, sodium hydrogen phosphate, NaOH, hydrochloric acid, phosphoric acid, cupric sulfate pentahydrate and basic copper chloride etc. are chemically pure reagent.
2. test method
The preparation of A, buffer solution: secure ph be respectively 2.0,3.0,4.0,5.0,6.0,7.0 phosphate buffer (PBS, 0.1M).
The mensuration of B, equilbrium solubility: getting cupric glutamate, to add the pH value respectively be in 2.0,3.0,4.0,5.0,6.0,7.0 the 100ml PBS solution; 60 ℃ of water-baths heating and ultrasonic is no longer dissolved to cupric glutamate; Put into water bath chader, temperature keeps (37 ± 1) ℃, jolting 24h; After saturated solution centrifugation,, calculate the equilbrium solubility of cupric glutamate with the copper ion in the aas determination solution; Same procedure is measured the equilbrium solubility of cupric sulfate pentahydrate, basic copper chloride.
3. result of the test
The result shows that cupric sulfate pentahydrate is soluble in water, and basic copper chloride is water insoluble, and acid condition is prone to down dissolve, and glutamic acid ketone is water-soluble low, and acidproof endurance strong (table 4).
The equilbrium solubility of table 4 cupric glutamate, copper sulphate and basic copper chloride
(6) the profit distribution coefficient of the cupric glutamate that obtains of step (1) is measured
1. test material: n-octyl alcohol, cupric glutamate, cupric sulfate pentahydrate, basic copper chloride
2. test method is an example with the cupric glutamate, and cupric sulfate pentahydrate, basic copper chloride are equally in order to method detection down
A, cupric glutamate are measured in the equilbrium solubility of n-octyl alcohol solution: take by weighing 5 gram cupric glutamates; Add 100 milliliters of n-octyl alcohol solution, 60 ℃ of water-baths heating and ultrasonic is no longer dissolved to cupric glutamate, puts into water bath chader; Temperature keeps (37 ± 1) ℃, jolting 24h; After saturated solution centrifugation,, calculate the equilbrium solubility of cupric glutamate in n-octyl alcohol solution with the copper ion in the aas determination solution.
B, vortex time are investigated: getting mass concentration is 400mgL
-1Cupric glutamate n-octyl alcohol solution (by water saturated after n-octyl alcohol) 0.5mL, add by n-octyl alcohol the 0.1molL after saturated
-1Hydrochloric acid solution 5ml, difference vortex 5,10,20,45,60min, 10000rmin
-1Centrifugal 2min gets the n-octyl alcohol layer is measured copper ion in the n-octyl alcohol with atom absorption method content; Confirm the righttest vortex time.
C, profit distribution coefficient are measured: n-octyl alcohol solution (saturated by the water) 0.5mL that gets cupric glutamate adds entry-cushioning liquid (saturated by n-octyl alcohol) 5ml as water as oil phase, vortex (result according to step B is fixed), 10000rmin
-1Centrifugal 2min, the oil phase 0.1ml that gets the upper strata measures copper ion concentration with atom absorption method, calculates the distribution situation of cupric glutamate at oil phase and aqueous phase, calculates profit and divides distribution coefficient (P)=Po/Pw.
3. result of the test
Solubility copper sulphate in water is the highest, and basic copper chloride takes second place, and cupric glutamate is minimum.The solubility of three's solubility copper sulphate is the highest in n-octyl alcohol solution, and cupric glutamate takes second place, and basic copper chloride is minimum.The profit distribution coefficient that calculates each compound is respectively: cupric glutamate 0.0102, basic copper chloride 0.001, and copper sulphate has only 0.0002.Cupric glutamate is than the antibacterial activity of basic copper chloride and copper sulphate high 10 times and 50 times (table 5), and profit divides the system of disposition high more, compound fat-soluble strong more, and antibacterial activity is then strong more.
The profit distribution coefficient of table 5 cupric glutamate, copper sulphate and basic copper chloride
(6) the growth promotion effect test of the cupric glutamate that obtains of step (1)
1. test material
Experimental animal: 350 the 1 yellow fast big fryer (herding research institute of guangdong agricultural science institute) in the age in days south of the Five Ridges; 50 weanling pigs (Guangdong the people live in plenty plant the pig farm);
Feed: the 102 type chickens that do not contain any antibacterials are used perfect compound feed, and Animal Husbandry Inst., Guangdong Prov. Academy of Agricultural Sciences's feed factory is special; The 303 type pigs that do not contain any antibacterials are used perfect compound feed, and the people live in plenty in Guangdong, and herding Development Co., Ltd feed factory is special;
Given the test agent: cupric glutamate, cupric sulfate pentahydrate.
2. test method
A, cupric glutamate are tested the growth promotion of the yellow fast big fryer in the south of the Five Ridges
350 the 1 yellow fast large-scale fryer in the age in days south of the Five Ridges are divided into 7 groups, 50 every group at random.Each group is added different growth promoters (table 6) in feed after, free choice feeding, the weightening finish and the price of deed of each test group test chicken of statistics 1～21 age in days, and relatively copper sulphate and cupric glutamate to the growth enhancing effect of fryer.
Table 6 cupric glutamate divides into groups to the growth promotion test of fryer
| Group | Size of animal | Average initial weight (g) | Growth promoter | Concentration (ppm) * |
| Do not add short long agent control group | 50 | 50 | - | - |
| Cupric sulfate pentahydrate-150 | 50 | 50 | Cupric sulfate pentahydrate | 150 |
| Cupric sulfate pentahydrate-100 | 50 | 50 | Cupric sulfate pentahydrate | 100 |
| Cupric sulfate pentahydrate-50 | 50 | 50 | Cupric sulfate pentahydrate | 50 |
| Cupric glutamate-150 | 50 | 50 | Cupric glutamate | 150 |
| Cupric glutamate-100 | 50 | 50 | Cupric glutamate | 100 |
| Cupric glutamate-50 | 50 | 50 | Cupric glutamate | 50 |
*: the working concentration of different growth promoters is in the copper ion in the compound.
B, cupric glutamate are tested the growth promotion of pig
60 weanling pigs such as table 7 divide into groups, 10 every group.Each group is added different growth promoters (seeing table 7) in feed after, free choice feeding, the weightening finish and the price of deed of back 30 days each test group test pig of statistics wean, and relatively copper sulphate and cupric glutamate to the growth enhancing effect of pig.
Table 7 cupric glutamate divides into groups to the growth promotion test of pork pig
| Group | Size of animal | Average initial weight (kg) | Growth promoter | Concentration (ppm) * |
| Do not add short long agent control group | 10 | 8.40 | ?- | - |
| Cupric sulfate pentahydrate-250 | 10 | 8.45 | Cupric sulfate pentahydrate | 250 |
| Cupric glutamate-100 | 10 | 8.52 | Cupric glutamate | 100 |
| Cupric glutamate-75 | 10 | 8.48 | Cupric glutamate | 75 |
| Cupric glutamate-50 | 10 | 8.25 | Cupric glutamate | 50 |
| Cupric glutamate-30 | 10 | 8.36 | Cupric glutamate | 30 |
*: the working concentration of different growth promoters is in the copper ion in the compound.
3. result of the test
A, cupric glutamate are tested the growth promotion of the yellow fast big fryer in the south of the Five Ridges
In the feeding experiment process of the yellow fast big fryer in the south of the Five Ridges; The copper sulphate group especially copper ion 150ppm and 100ppm group in second week of test; Poisoning symptoms such as feather is in disorder, skin is dry appear in the yellow fast big fryer in the south of the Five Ridges; A small amount of dead chicken phenomenon before off-test, also occurs, explain that copper sulphate is added on the medium-term and long-term use of feed at 100ppm and the copper ion poisoning can occur.Though clinical visible poisoning symptom does not appear in the copper sulphate group of 50ppm copper ion, the relative weight gain rate of experimental period is 99.6%, and is approaching with not administration control group, but feedstuff-meat ratio than control group high 0.052.3 dose groups of cupric glutamate all show good growth promoting function, and the relative weight gain rate can improve 6.3-7.3%, the price of deed 0.054-0.071 (seeing table 8 for details) that can descend.Security that growth test results suggest cupric glutamate tool is good and growth promotion effect.
Table 8 cupric glutamate is to the growth promotion result of the test of fryer
B, cupric glutamate are tested the growth promotion of pig
In the swine rearing process of the test, the cupric sulfate pentahydrate group (250ppm) of high dose has tangible growth promoting function, and flat more not administration of the counterpoise control group that increases of duration of test improves 7.4%, and the price of deed reduces by 0.177.The cupric glutamate of various dose group all shows the growth enhancing effect with high dose copper sulphate, and the growth promotion effect close (table 9) of 50ppm and 100ppm and 150ppm dose groups, and the cupric glutamate of results suggest 50ppm can replace the copper sulphate of 250ppm.
Table 9 cupric glutamate is to the growth promotion result of the test of pig
Embodiment 2
The cupric glutamate derivative is to the growth promotion effect test of animal
(1) preparation of the derivative of different cupric glutamates
A, cupric glutamate hydrochloride: take by weighing 100 gram CuSO
45H
2O and 74.8 gram sodium glutamates are dissolved in respectively in the 1000ml water, stir to dissolving fully, and the two is mixed, and with the 0.1M hydrochloric acid solution pH value of mixed solution are transferred to 4.0, and heating and continuous stirring are reacted, and gather in the crops crystallization and dry to constant weight.Get product and be dissolved in water with 1 gram and be settled to 100ml, measure the content of copper ion (atomic absorption method), chlorion (Moire technique) and glutamic acid (amino-acid analyzer) in the solution.Copper ion concentration is 2.607mg/ml as a result, and chloride ion content is 1.446mg/ml, and the content of glutamic acid is 5.908mg/ml, and the crystal that can infer gained thus is the cupric glutamate hydrochloride that cupric glutamate and hydrochloric acid formed in 1: 1 in molar ratio.
The preparation of B, glutamic acid methyl ester copper chelate: take by weighing 100 gram CuSO
45H
2O and 129.0 gram glutamic acid methyl esters are dissolved in respectively in the 1000ml water, stir to dissolving fully, with the two mixing, with 1M sodium hydroxide solution seasoning pH value to 7.0, centrifugal collecting precipitation, and behind the water centrifuge washing three times 100 ℃ dry to constant weight.Get and be settled to 100ml water, the content of copper ion (atomic absorption method) and glutamic acid (amino-acid analyzer) in the mensuration solution after 1 gram product dissolves with 0.01M HCl solution fully.Copper ion concentration is 1.592mg/ml as a result, and the content of glutamic acid is 7.314mg/ml, and the product that can infer gained thus is the chelate of band a part crystallization water of being formed in 2: 1 in molar ratio by glutamic acid methyl ester and copper ion.
The preparation of C, N-carbamylglutamic acid copper: take by weighing 100 gram CuSO
45H
2O and 76.8 gram N-carbamylglutamic acids are dissolved in respectively in the 1000ml water, stir to dissolving fully, with the two mixing, with 1M sodium hydroxide solution seasoning pH value to 7.0, centrifugal collecting precipitation, and behind the water centrifuge washing three times 100 ℃ dry to constant weight.Get and be settled to 100ml water, the content of copper ion (atomic absorption method) and glutamic acid (amino-acid analyzer) in the mensuration solution after 1 gram product dissolves with 0.01M HCl solution fully.Copper ion concentration is 2.215mg/ml as a result, and the content of glutamic acid is 5.087mg/ml, and the product that can infer gained thus is the chelate of band a part crystallization water of being formed in 1: 1 in molar ratio by N-carbamylglutamic acid and copper ion.
The preparation of D, glutamine copper: take by weighing 100 gram CuSO
45H
2O and 116.8 gram glutamine are dissolved in respectively in the 1000ml water, stir to dissolving fully, with the two mixing, with 1M sodium hydroxide solution seasoning pH value to 7.0, centrifugal collecting precipitation, and behind the water centrifuge washing three times 100 ℃ dry to constant weight.Get and be settled to 100ml water, the content of copper ion (atomic absorption method) and glutamine (amino-acid analyzer) in the mensuration solution after 1 gram product dissolves with 0.01M HCl solution fully.Copper ion concentration is 1.721mg/ml as a result, and the content of glutamic acid is 7.850mg/ml, and the product that can infer gained thus is the chelate of band a part crystallization water of being formed in 2: 1 in molar ratio by N-carbamylglutamic acid and copper ion.
The preparation of E, lysyl cupric glutamate: take by weighing 100 gram CuSO
45H
2O and 110.0 gram lysyl glutamic acid are dissolved in respectively in the 1000ml water, stir to dissolving fully, with the two mixing, with 1M sodium hydroxide solution seasoning pH value to 7.0, centrifugal collecting precipitation, and behind the water centrifuge washing three times 100 ℃ dry to constant weight.Get and be settled to 100ml water after 1 gram product dissolves with 0.01M HCl solution fully, measure the content of copper ion (atomic absorption method), lysine and glutamic acid (amino-acid analyzer) in the solution.Copper ion concentration is 1.803mg/ml as a result; Lysine content 4.113mg/ml; The content of glutamic acid is 4.141mg/ml, and the product that can infer gained thus is the chelate of band a part crystallization water of being formed in 1: 1 in molar ratio by lysyl glutamic acid and copper ion.
The preparation of F, salicylide cupric glutamate (cupric glutamate shiff alkali): 0.02mmol potassium hydroxide is dissolved in the 1000mL distilled water. and be added drop-wise to 500mL and contain in 0.02mmol sodium glutamate and the 0.02mmol salicylide ethanolic solution; Under 80 ℃ of water-baths; Stirring reaction 2h adds 1000mL then and contains 0.02mmol CuSO
45H
2The O aqueous solution is regulated pH to 7.0, continues reaction 8h, and cold filtration is collected heavy and dried to constant weight.Get and be settled to 100ml water, the content of copper ion (atomic absorption method) and glutamic acid (amino-acid analyzer) in the mensuration solution after 1 gram product dissolves with 0.01M HCl solution fully.Copper ion concentration is 1.835mg/ml as a result, and the content of glutamic acid is 4.200mg/ml, and the product that can infer gained thus is the chelate of band a part crystallization water of being formed in 1: 1 in molar ratio by salicylide glutamic acid and copper ion.
(2) the cupric glutamate derivative is to the promoting animal growth Research on effect
1. test material
Experimental animal: 90 weanling pigs: the people live in plenty in Guangdong plants the pig farm provides;
Feed: the 303 type pigs that do not contain any antibacterials are used perfect compound feed, and the people live in plenty in Guangdong, and herding Development Co., Ltd feed factory is special;
Given the test agent: six kinds of cupric glutamate derivatives (cupric glutamate hydrochloride, glutamic acid methyl ester copper chelate, N-carbamylglutamic acid copper, glutamine copper, lysyl cupric glutamate and salicylide cupric glutamate), the CuSO of cupric glutamate, step (1) preparation
45H
2O.
2. test method
90 weanling pigs such as table 9 divide into groups, 10 every group.Each group is added different growth promoters (seeing table 10) in feed after, free choice feeding, the weightening finish and the price of deed of back 30 days each test group test pig of statistics wean, and relatively copper sulphate and cupric glutamate or derivatives thereof to the difference of the growth enhancing effect of pig.
The different cupric glutamate derivatives of table 10 divide into groups to the growth promotion test of pork pig
| Group | Size of animal | Average initial weight (kg) | Growth promoter | Concentration (ppm) * |
| 1 | 10 | 8.32 | ?- | - |
| 2 | 10 | 8.35 | Cupric sulfate pentahydrate | 250 |
| 3 | 10 | 8.42 | Cupric glutamate | 50 |
| 4 | 10 | 8.40 | The cupric glutamate hydrochloride | 50 |
| 5 | 10 | 8.45 | The glutamic acid methyl ester copper chelate | 50 |
| 6 | 10 | 8.46 | Cupric glutamate | 50 |
| 7 | 10 | 8.30 | N-carbamylglutamic acid copper | 50 |
| 8 | 10 | 8.35 | The lysyl cupric glutamate | 50 |
| 9 | 10 | 8.36 | The salicylide cupric glutamate | 50 |
*: the working concentration of different growth promoters is in the copper ion in the compound.
3. the cupric glutamate derivative is to the growth promotion test result of the test of pork pig
Result of the test among the embodiment 1 has confirmed that there is suitable growth promoting function in the cupric glutamate copper source of 50ppm and the cupric sulfate pentahydrate copper source of 250ppm.The cupric sulfate pentahydrate copper source (group 2) of the cupric glutamate copper source of 50ppm in the present embodiment (group 3) and 250ppm has improved 11.36% and 10.63% respectively than the average weight gain of control group; Feedstuff-meat ratio has also reduced by 0.273 and 0.261 respectively, has reconfirmed that the cupric glutamate copper source of 50ppm in the feed process for preparation can substitute the cupric sulfate pentahydrate copper source of 250ppm.In addition, the various derivatives of cupric glutamate and paddy ammonia copper have suitable growth promotion effect (table 11), and the derivative of results suggest cupric glutamate and cupric glutamate have suitable biological effect.
The different cupric glutamate derivatives of table 11 are to the growth promotion result of the test of pig
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (3)
1. the cupric glutamate or derivatives thereof is characterized in that as the application of animal feed additive for promoting growth: the chelate that described derivative forms for dipeptides and copper ion with a kind of formation in glutamic acid and the amino acid.
2. according to the application of the said cupric glutamate or derivatives thereof of claim 1 as animal feed additive for promoting growth, it is characterized in that: described animal is a cultivated animals.
3. according to the application of the said cupric glutamate or derivatives thereof of claim 1 as animal feed additive for promoting growth, it is characterized in that: said cupric glutamate or derivatives thereof is applied to each growth phase of animal.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113170837A (en) * | 2021-04-07 | 2021-07-27 | 彭险峰 | Copper aspartate complex and application thereof |
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| CN1467198A (en) * | 2002-07-12 | 2004-01-14 | 上海福赐德营养保健品有限责任公司 | Monozinc di-L-Aspartate |
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| CN113170837A (en) * | 2021-04-07 | 2021-07-27 | 彭险峰 | Copper aspartate complex and application thereof |
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