CN103772528B - A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder - Google Patents
A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder Download PDFInfo
- Publication number
- CN103772528B CN103772528B CN201410012934.7A CN201410012934A CN103772528B CN 103772528 B CN103772528 B CN 103772528B CN 201410012934 A CN201410012934 A CN 201410012934A CN 103772528 B CN103772528 B CN 103772528B
- Authority
- CN
- China
- Prior art keywords
- chitosan
- selenium
- sulfuric ester
- sulfated
- sodium selenite
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The method of the water-soluble selenizing sulfated chitosan of a kind of quick production, by chitosan solution and sulfonated reagent mixing are synthesized sulfated chitosan, again sulfated chitosan and selenium Standard Stock solutions are mixed in proportion, stirring reaction 0.5-4h under 10-60 DEG C of condition, 95% alcohol settling, washing, oven drying at low temperature obtains faint yellow selenium chitosan sulfuric ester.The product obtained is soluble in water, selenium content and sulfate radical content is high, stable in properties, the growth performance of chicken, liver function and anti-oxidant function can be improved, in chicken body, there is good biological function, be with a wide range of applications in aquaculture as additive for farm animal feed.
Description
Art
The invention belongs to the technical field of selenizing Sulfate of polysaccharide, relate to a kind of water-soluble preparation method of selenizing sulfated chitosan and the application in animal and fowl fodder.
Background technology
Chitosan (chitosan; CTS) be chitin (chitin) product after deacetylated process; i.e. Chitosan; formal name used at school poly-glucosamine, existing oneself is called isolating protein by many scientists, fat; sugar; VITAMIN, necessary 6th vital principle of human body outside mineral substance, has the effects such as strengthening immune, anti-ageing, anti-oxidant, anticancer, rheumatism and prevents AIDS.The chitosan of the natural organic compounds containing nitrogen maximum as occurring in nature quantity and second largest organic natural resources, because himself has physiological compatibility, can decomposability completely, polyfunctional reactant, three-dimensional arrangement and chirality, the multiple excellent properties such as recyclability and wetting ability makes the research of chitosan apply and is more and more subject to worldwide extensive concern.Chitosan obtains sulfated chitosan after sulphation modification, its antiviral, antitumor and strengthen immunity etc. biologic activity significantly improve.The combination of selenium and chitosan can optimize respective effect, makes its antioxygenation more remarkable.Containing-OH and-NH2 active group in chitosan molecule, it is the ideal carrier of various trace elements and chemical group.Chitosan, through oversulfonate or selenizing, introduces SO42-group or selenium, obtains sulfated chitosan or chitosan selenium acid esters, enhances the biologic activity of chitosan.But, chitosan and compound water soluble poor, although can be dissolved in acidic solution, easily there is Swelling, be chitosan compound synthesis in a difficult problem, limit the widespread use of chitosan in livestock industry field.
Selenium (Selenium, Se) be one of required trace element of body, have anti-oxidant, anti-cancer, removing toxic substances, anti-stress effect, also certain promoter action is had to the Growth and reproduction of animal, appropriate selenium is supplemented in feed and drinking-water, the immunological competence of animal body can be improved, and can prevention and therapy low selenium area endemicity selenium deficiency.Selenium insufficiency of intake can cause body health and have a strong impact on, and even can cause various diseases, as Keshan disease, Kaschin-Beck disease, the white muscle disease of cattle and sheep, the exudative diathesis of bird, the breeding difficulty etc. of animal of people.At present, Selenium Supplementation In Diet has become current cultivation industry prevention selenium deficiency, has strengthened important measures of animal resistibility and raising reproductive performance.Current conventional selenium source comprises inorganic selenium and organoselenium, and Sodium Selenite (Na2SeO3) is the most frequently used inorganic selenium source, and its safety range is little, and bioavailability is low, comparatively serious to the pollution of environment and soil.Compared with inorganic selenium, organoselenium has the advantages such as toxicity is little, bioavailability is high, environmental pollution is little, and development of new organoselenium preparation has become the focus of current research.
Selenium chitosan is the organic compound of selenium and chitosan, the dual-use function of selenium and chitosan can be played simultaneously, have and improve broiler chicken production performance, improve immunity of organisms, promote bifidus bacillus and lactic acid bacterium number in intestinal microflora, reduce the effect of escherichia coli quantity.In addition, chitosan selenium can improve the anti-oxidant function of mouse, reduces plumbous cumulative toxicity.
China also exists and lacks selenium area widely, and mending selenium has been the conventional measure that domestic livestock industry is taked.In view of the pollution that inorganic selenium toxicity is comparatively strong, cause to soil and environment and have potential danger and the particular advantages of organoselenium to human health, application organoselenium Selenium supplement agent replaces inorganic selenium to be trend of the times.At present, organic selenium source is mainly yeast rich in selenium and Sethotope, and price is more expensive, is several times even tens times of Sodium Selenite price.Therefore, produce selenium content high, low-cost organoselenium preparation is livestock industry problem demanding prompt solution now.
In recent years, the patent of selenium chitosan sulfuric ester occurs less.As: chitosan selenite and preparation method thereof (CN1284509A), the method of the chitosan selenite described in it, by the aqueous solution of chitosan and tin anhydride, after the stirring reaction 1-3h of interval, add alcohol settling, filter, remove unreacted selenous acid by washing with alcohol, after drying, namely obtain chitosan selenite product.Use the reagent such as selenous acid and acetone in this preparation method, pollute larger.Patent polysaccharide sulfate selenide compound and method for making thereof and application (publication number: CN1011046C4643A), by Sulfate of polysaccharide by heating 10-100 DEG C, isothermal reaction 1-100h generates selenizing Sulfate of polysaccharide, then using sulfated solution removing barium ion, add ferric chloride Solution after filtration, after adjustment pH, stirring reaction spends the night, and removes unreacted selenous acid or salt, then apply alcohol settling, much filtrate is dry.This method is prepared loaded down with trivial details, and the time is longer, uses the corrosive liquid such as sulfuric acid, pollutes larger.
Summary of the invention
For prior art Problems existing, the invention provides method and this selenium chitosan sulfuric ester application in animal and fowl fodder of the water-soluble selenizing sulfated chitosan of a kind of quick production
The concrete technical scheme that the present invention adopts is as follows:
The first step: add dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips chlorsulfonic acid, at room temperature stirs, obtain sulfonated reagent after dropwising;
Second step: by dichloro acetic acid and methane amide mixing, add chitosan and stir, obtaining chitosan solution;
3rd step: Sodium Selenite is dissolved in distilled water, and constant volume, obtain the selenium Standard Stock solutions that selenium content is 1gL – 1;
4th step: the synthesis of sulfated chitosan: by chitosan solution and sulfonated reagent mixing, reaction 0.5-4h to reactant become brown after, by reactant impouring dehydrated alcohol, low temperature places 0.5-4h, forms sulfated chitosan crude product; Precipitation, through vacuum filtration, after washing with alcohol 1-2 time, carries out oven drying at low temperature;
5th step: selenium chitosan sulfuric ester synthesizes: be dissolved in by sulfated chitosan in a small amount of water, regulator solution pH is 3-7, constant volume.According to sulfated chitosan: Sodium Selenite=1: sulfated chitosan and selenium Standard Stock solutions mix by the mass ratio of 1-1: 10, stirring at room temperature 0.5-4h.Add 3-5 times amount 95% alcohol settling 0.5-4h, precipitate through vacuum filtration, 95% washing with alcohol, oven drying at low temperature, obtain faint yellow selenium chitosan sulfuric ester.
By application TU-1901 twin-beam ultraviolet-visible pectrophotometer, in the scanning of 600-190nm interval, draw spectroscopic analysis curve, detected result is shown in accompanying drawing 1-3.Having there is absorption peak at 194nm place in selenium chitosan sulfuric ester, illustrates to there occurs selenylation reaction on sulfated chitosan.
Adopt the method for sample and pressing potassium bromide troche, application THEMONICOLETAVATAR360 type FT-TR spectrograph, in the scanning of 400-4000cm-1 interval, result as shown in Figure 4.Find that selenium chitosan sulfuric ester obviously weakens at the absorption peak of 3434cm-1 position, illustrate that N-H radical moiety on C2 position is by selenite radical ionic replacement, the absorption peak in 1237cm-1 site obviously strengthens, and the absorption peak at 1232cm-1 site and 809cm-1 place presents the trend weakened, there is the Se=O double bond vibration peak of monohydrate selenium dioxide ester in 813cm-1 site, the reaction that C3 and C6 position all there occurs part selenite radical replacement sulfate group has been described.
The preparation method of quick production provided by the invention water-soluble selenizing sulfated chitosan has following beneficial effect:
The synthetic method of selenium chitosan sulfuric ester of the present invention is simple and quick, and generated time, between 4-6h, simplifies middle multiple steps, simple compared with other synthetic method step, and generated time is short.
The present invention prepares the method for selenium chitosan sulfuric ester and additive method ratio, employs less chemical reagent, avoids the use of number of chemical reagent, decreases the number of chemical refuse produced in building-up process.
Product is soluble in water.Carry out according to GB/T21845-2008 method, take the selenium chitosan sulfuric ester 0.1g being ground into fine powder and add in the graduated cylinder with glass ground joint plug, as 20 DEG C ± 0.5 DEG C, after adding 0.1ml distilled water, observe dissolving situation at every turn.Found that, namely occur to dissolve completely after 0.1g selenium chitosan sulfuric ester adds 1ml distilled water, show that selenium chitosan sulfuric ester solubleness in water is greater than 1g/L, soluble in water.
Selenium content and the sulfate radical content of product are higher, and selenium content is 31.60-75.60mgg – 1, sulfate radical content is 213.68-332.8mgg – 1.
Selenium content and the sulfate radical content of product are stablized, and the impact by solution acid alkalinity, time, temperature is less, stable in properties.Selenium chitosan sulfuric ester powder to be placed in constant temperature oven 60 DEG C to place 10 days, to detect and find that selenium content and sulfate radical content are all in more than 95% of original concentration.By selenium chitosan sulfuric ester powder and water-soluble product respectively as in normal temperature air and in 4 DEG C of refrigerators, measured after 1 year and find, selenium content and sulfate radical content are all in more than 90% of original concentration.
The hepatocellular antioxygenation of the selenium chitosan sulfuric ester external raising of energy of the present invention, production performance and the anti-oxidant function of chicken can be improved, play the double biological effect of selenium and sulfuric ester, therefore selenium chitosan sulfuric ester of the present invention can improve the growth performance of chicken, in chicken body, there is good biological function, be with a wide range of applications in aquaculture as additive for farm animal feed.
After liver cell being inoculated 6 porocyte culture plates cultivation 24h, chitosan selenium, selenium chitosan sulfuric ester, Sodium Selenite (in selenium) and sulfated chitosan, chitosan (in the chitosan amount that selenium is corresponding) is added in nutrient solution, cultivate and to get the mensuration that cell conditioned medium liquid and cell pyrolysis liquid carry out Antioxidant Indexes after 24h, detect the expression amount of cellularity glutathione peroxidase (GPx1) and human phospholipid hydroperoxide glutathione peroxidase (GPx4) mRNA in liver cell simultaneously.Found that, selenium concentration is 5 μm of olL – 1 selenium chitosan sulfuric ester group liver cell POD activity is 13.61 ± 0.71, is significantly higher than chitosan group (11.31 ± 0.61) and blank group (9.92 ± 1.78); Total antioxidant capacity is 11.18 ± 0.99, is significantly higher than chitosan group (4.07 ± 0.44) and blank group (2.92 ± 0.68); Superoxide anion resisitance free radical activity is 193.01 ± 23.39, is significantly higher than blank group (95.50 ± 6.34); Catalase activity is 4.91 ± 0.15, is significantly higher than Sodium Selenite group (3.96 ± 0.20), chitosan group (4.02 ± 0.59) and blank group (3.30 ± 0.25); GPx1mRNA expression amount is 9.06 ± 1.46, is significantly higher than Sodium Selenite group (0.99 ± 0.17), chitosan selenium group (2.49 ± 0.40), chitosan group (1.81 ± 0.32) and blank group (1.30 ± 0.62); GPx4mRNA expression amount is 6.47 ± 1.27, is significantly higher than Sodium Selenite group (0.63 ± 0.04), chitosan selenium group (3.10 ± 0.30), chitosan group (0.74 ± 0.05) and blank group (1.76 ± 0.86).Illustrate that selenium chitosan sulfuric ester has good antioxygenation to liver cell, effect is better than chitosan, Sodium Selenite and chitosan selenium.
Choose the healthy SPF laying hen of 60 1 ages in days, after raising 7d, be divided into 4 groups at random, often organize 15.To feed respectively basal diet, Sodium Selenite, chitosan selenium and selenium chitosan sulfuric ester.Test to 28d and measure the mrna expression amount of the net weight of chicken, average daily gain, blood plasma Antioxidant Indexes, liver function index and liver GPx1 and GPx4.Found that, the 28d of test, selenium chitosan sulfuric ester group SPF chicken body weight reaches 581 ± 55.7g, is significantly higher than control group (465.00 ± 9.54g), Sodium Selenite group (481.4 ± 31.24g), chitosan selenium group (469.2 ± 64.86); Selenium chitosan sulfuric ester group SPF chicken average daily gain 17.14g, is significantly higher than control group (13g), Sodium Selenite group (13.57g), chitosan selenium group (13.18).Selenium chitosan sulfuric ester group SPF chicken plasma total antioxidant capacity is 7.4 ± 0.39, is significantly higher than control group (1.97 ± 0.21), Sodium Selenite group (5.67 ± 0.98), chitosan selenium group (5.42 ± 0.45); Selenium chitosan sulfuric ester group SPF chicken plasma GSH-PX activity is 775.97 ± 40.65, is significantly higher than control group (660.73 ± 84.08), Sodium Selenite group (729.02 ± 64.45); Selenium chitosan sulfuric ester group SPF chicken plasma superoxide dismutase activity is 163.73 ± 4.32, is significantly higher than control group (107.55 ± 15.16), Sodium Selenite group (120.41 ± 13.34), chitosan selenium group (136.68 ± 11.92); Selenium chitosan sulfuric ester group SPF chicken plasma mda content is 2.20 ± 0.15, is significantly higher than control group (3.18 ± 0.0.37), Sodium Selenite group (2.84 ± 0.16), chitosan selenium group (3.04 ± 0.22).Selenium chitosan sulfuric ester group SPF chicken plasma Pancreas Glutamate-Pyruvate Transaminase Vigor is 212.84 ± 1.05, is significantly higher than control group (182.22 ± 2.57).Test 14d, selenium chitosan sulfuric ester group SPF chicken liver GPx1mRNA expression amount is 6.50 ± 1.78, is significantly higher than control group (0.93 ± 0.05), Sodium Selenite group (4.28 ± 0.45), chitosan selenium group (2.57 ± 0.29); Selenium chitosan sulfuric ester group GPx4mRNA expression amount is 3.00 ± 0.67, is significantly higher than control group (1.13 ± 0.09), Sodium Selenite group (2.14 ± 0.14), chitosan selenium group (2.48 ± 0.36).Illustrate that selenium chitosan sulfuric ester can improve the growth performance of chicken, liver function and resistance of oxidation.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the present invention is further described:
The UV spectrum collection of illustrative plates of Fig. 1 chitosan
The UV spectrum collection of illustrative plates of Fig. 2 sulfated chitosan
The UV spectrum collection of illustrative plates of Fig. 3 selenium chitosan sulfuric ester
The infrared spectra collection of illustrative plates of Fig. 4 chitosan
The infrared spectra collection of illustrative plates of Fig. 5 sulfated chitosan
The infrared spectra collection of illustrative plates of Fig. 6 selenium chitosan sulfuric ester
Embodiment
Embodiment 1
1, add 50ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 10ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 1h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, 60 DEG C, react 1h under 100rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 20 DEG C, form sulfated chitosan crude product; Precipitation, through vacuum filtration, after washing with alcohol 2 times, carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 164%, and sulfate radical content is 406.8mgg – 1.
5, selenium chitosan sulfuric ester synthesis: take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 0.5h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 35.34mgg – 1, sulfate radical content is 282.24mgg – 1.
Embodiment 2
1, add 100ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 30ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 2h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, at 60 DEG C, reacts 1h under 100rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 20 DEG C, form sulfated chitosan crude product; Precipitation is through vacuum filtration, and after washing with alcohol 2 times, product carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 159%, and sulfate radical content is 394.6mgg – 1.
5, selenium chitosan sulfuric ester synthesis
(1) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 0.5h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 36.50mgg – 1, sulfate radical content is 259.81mgg – 1.
(2) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 33.61mgg – 1, sulfate radical content is 264.9mgg – 1.
(3) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 2h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 33.14mgg – 1, sulfate radical content is 287.8mgg – 1.
(4) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 4h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 32.07mgg – 1, sulfate radical content is 279.53mgg – 1.
Embodiment 3
1, add 100ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 30ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 1h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, at 60 DEG C, reacts 2h under 120rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 20 DEG C, form sulfated chitosan crude product; Precipitation is through suction filtration, and after washing with alcohol 2 times, product carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 149%, and sulfate radical content is 387.8mgg – 1.
5, selenium chitosan sulfuric ester synthesis:
(1) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 20 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 41.46mgg – 1, sulfate radical content is 273.36mgg – 1.
(2) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 43.95mgg – 1, sulfate radical content is 276.22mgg – 1.
(3) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 40 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 43.07mgg – 1, sulfate radical content is 276.07mgg – 1.
(4) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 50 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 42.27mgg – 1, sulfate radical content is 274.87mgg – 1.
(5) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 60 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 40.93mgg – 1, sulfate radical content is 281.79mgg – 1.
Embodiment 4
1, add 100ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 30ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 2h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, at 40 DEG C, reacts 2h under 120rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 20 DEG C, form sulfated chitosan crude product; Precipitation is through suction filtration, and after washing with alcohol 2 times, product carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 157.2%, and sulfate radical content is 446.9mgg – 1.
5, selenium chitosan sulfuric ester synthesis:
(1) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 43.96mgg – 1, sulfate radical content is 276.2mgg – 1.
(2) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 4, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 38.91mgg – 1, sulfate radical content is 261.50mgg – 1.
(3) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 5, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 37.30mgg – 1, sulfate radical content is 272.28mgg – 1.
(4) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 6, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 37.27mgg – 1, sulfate radical content is 269.97mgg – 1.
Embodiment 5
1, add 100ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 30ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 1h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, at 10 DEG C, reacts 1h under 120rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 10 DEG C, form sulfated chitosan crude product; Precipitation is through suction filtration, and after washing with alcohol 2 times, product carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 152%, and sulfate radical content is 378.2mgg – 1.
5, selenium chitosan sulfuric ester synthesis:
(1) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 1 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 75.60mgg – 1, sulfate radical content is 213.68mgg – 1.
(2) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 2 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 65.18mgg – 1, sulfate radical content is 229.15mgg – 1.
(3) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 3 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 59.44mgg – 1, sulfate radical content is 236.87mgg – 1.
(4) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 5 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 54.29mgg – 1, sulfate radical content is 232.57mgg – 1.
(5) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 10 mixes, under 30 DEG C of conditions, stir 1h.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 31.60mgg – 1, sulfate radical content is 254.74mgg – 1.
Embodiment 6
1, add 100ml dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 30ml chlorsulfonic acid, at room temperature stirs 30min, obtain sulfonated reagent after dropwising;
2, by 3.5ml dichloro acetic acid and the mixing of 50ml methane amide, add 3g chitosan and stir 1h, obtain chitosan solution;
3, take 2.190g Sodium Selenite and be dissolved in distilled water, be settled to 1L, containing selenium 1gL – 1, obtain selenium Standard Stock solutions;
4, the synthesis of sulfated chitosan: mix according to chitosan 3.0g and sulfonated reagent 60ml reaction system, at 10 DEG C, reacts 1h under 120rmin – 1 condition, reactant becomes brown, in impouring 5 times of dehydrated alcohols, place 2h for 10 DEG C, form sulfated chitosan crude product; Precipitation is through suction filtration, and after washing with alcohol 2 times, product carries out 60 DEG C of dryings; The productive rate of the sulfated chitosan of preparation is 162%, and sulfate radical content is 398.4mgg – 1.
5, selenium chitosan sulfuric ester synthesis:
(1) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 1 mixes, under 30 DEG C of conditions, stir 45min.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 35.3mgg – 1, sulfate radical content is 232.76mgg – 1.
(2) take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 4, is settled to 25ml.According to Sodium Selenite: sulfated chitosan=1: the ratio of 3 mixes, under 40 DEG C of conditions, stir 15min.Cooling, adds 4 times of alcohol settling 2h, and through vacuum filtration, washing, dries to obtain selenium chitosan sulfuric ester for 60 DEG C.Selenium content is 39.21mgg – 1, sulfate radical content is 213.81mgg – 1.
Claims (2)
1. a preparation method for selenium chitosan sulfuric ester, is characterized in that, comprises the following steps:
(1) add 50-100mL dimethyl formamide to in the there-necked flask of whipping appts, ice bath drips 10-30mL chlorsulfonic acid, at room temperature stirs 10-30min, obtain sulfonated reagent after dropwising;
(2) by 2-10mL dichloro acetic acid and the mixing of 25-100mL methane amide, add 2-10g chitosan and stir 0.5-2h, obtain chitosan solution;
(3) take quantitative Sodium Selenite and be dissolved in distilled water, distilled water is settled to containing selenium 1gL
-1, obtain selenium Standard Stock solutions;
(4) synthesis of sulfated chitosan: according to chitosan 3.0g and the mixing of sulfonated reagent 60mL reaction system, 10-60 DEG C, 50-120rmin
-1react 0.5-4h under condition, reactant becomes brown, in impouring 3-5 times of 95% ethanol, places 0.5-4h for 0-40 DEG C, forms sulfated chitosan crude product; Precipitation, through vacuum filtration, after washing with alcohol 1-2 time, carries out 40-60 DEG C of oven dry;
(5) selenium chitosan sulfuric ester synthesis: take 5.0g sulfated chitosan, be dissolved in a small amount of water, regulator solution pH is 3.0-7.0, be settled to 25mL, according to Sodium Selenite: sulfated chitosan=1: the mass ratio of 1-1: 10 is by sulfated chitosan and the mixing of selenium Standard Stock solutions, stirring reaction 0.5-4h under 10-60 DEG C of condition, add 3-5 times amount 95% alcohol settling 0.5-4h, precipitation is through vacuum filtration, 95% washing with alcohol, 40-60 DEG C of oven dry, obtains faint yellow selenium chitosan sulfuric ester.
2. the selenium chitosan sulfuric ester prepared of a preparation method as claimed in claim 1 is as the application of additive in animal and fowl fodder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410012934.7A CN103772528B (en) | 2014-01-13 | 2014-01-13 | A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410012934.7A CN103772528B (en) | 2014-01-13 | 2014-01-13 | A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103772528A CN103772528A (en) | 2014-05-07 |
| CN103772528B true CN103772528B (en) | 2016-03-30 |
Family
ID=50565337
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410012934.7A Expired - Fee Related CN103772528B (en) | 2014-01-13 | 2014-01-13 | A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103772528B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108822232A (en) * | 2018-05-03 | 2018-11-16 | 青岛大学 | A kind of preparation method of cholic acid grafting selenizing chitosan oligosaccharide |
| CN109453207B (en) * | 2018-12-30 | 2021-12-31 | 广东南芯医疗科技有限公司 | Sodium selenylation alginate and chitosan selenylation coated probiotic double-layer microcapsule, preparation method and application thereof |
| CN111087484B (en) * | 2019-07-15 | 2020-08-25 | 郑州市御合源生物科技有限公司 | Bonded selenium polysaccharide and preparation method and application thereof |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1284509A (en) * | 2000-07-14 | 2001-02-21 | 武汉大学 | Chitosan selenite and its preparation |
| KR100814012B1 (en) * | 2004-06-24 | 2008-03-14 | 정한춘 | Manufacturing process of Chitosan-Selenium Complex |
| WO2007058396A1 (en) * | 2005-11-16 | 2007-05-24 | Han Chun Jung | Chitosan-selenium composite and method for preparing thereof |
| CN101104644B (en) * | 2006-07-14 | 2010-07-21 | 陈艺新 | Preparation for polysaccharide selenite and application thereof |
| CN101104643A (en) * | 2006-07-14 | 2008-01-16 | 陈艺新 | Polysaccharide sulfate selenide compound and its preparation method and application |
| CN101851259A (en) * | 2009-04-02 | 2010-10-06 | 辽宁师范大学 | Selenium chitosan oligosaccharide and preparation method thereof |
-
2014
- 2014-01-13 CN CN201410012934.7A patent/CN103772528B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN103772528A (en) | 2014-05-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103772528B (en) | A kind of preparation method of selenium chitosan sulfuric ester and the application in animal and fowl fodder | |
| CN102181371B (en) | Yeast and method for producing selenium-enriched yeast by utilizing yeast | |
| CN102816707B (en) | Candida utilis strain and application of same in preparation of selenium-rich yeast products | |
| CN101870740B (en) | Morchellaconica extracellular polysaccharide extractive and preparation method and application thereof | |
| CN110642955B (en) | Esterified selenium polysaccharide and preparation method and application thereof | |
| CN101508961A (en) | Method of preparing yeast rich in copper | |
| CN104561211A (en) | Method for improving beta-carotene produced from blakeslea trispora by virtue of organic acid | |
| CN105039236A (en) | Inoculating culture and detecting method for gathering copper ions through saccharomycete | |
| CN107586742A (en) | The bacillus amyloliquefaciens of one plant height production levulan and its application | |
| CN104764830A (en) | Method for detecting sulfonamide antibiotic residues in milk | |
| Gaponov et al. | Influence of chlorella on hematological parameters and metabolism of rhesus monkeys | |
| CN104620853A (en) | Culture method of ganoderma lucidum spore powder or sporocarp rich in selenium | |
| CN104711203A (en) | Selenium-rich germanium yeast powder | |
| CN110257296A (en) | The cultural method and application thereof of chromium-rich spirulina | |
| CN101797015A (en) | Preparation method of composite premix feed and obtained composite premix feed | |
| CN103130550B (en) | Culture medium and culture method of male agaric mycelium | |
| CN103406080B (en) | Complex coacervation prepares the CLA microcapsules that extracellular polysaccharide of bacteria is wall material | |
| CN102771836B (en) | Method for producing beta-carotene rich dunaliella powder | |
| CN107119038A (en) | A kind of Zinc-rich saccharomyces cerevisiae product and its production method | |
| CN105145115B (en) | A kind of production method of organic Se-rich zinc-rich product | |
| JP2024025666A (en) | Schizophyllum commune culture medium containing plant composition as well as preparation method and application of the same | |
| CN109880750A (en) | A kind of application of the method for improving Thelephora ganbajun mycelium and polysaccharide yield and wizened granulose | |
| CN106501191A (en) | The high throughput screening system of functional components of chlorella vulgaris CPE, screening technique and preparation method thereof | |
| Xiaoguang et al. | Improvement of Physiological Characteristic of Selenium‐Enriched Candida utilis with Amino Acids Addition | |
| CN105077218B (en) | A kind of preparation method of Selenium-enriched health food |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160330 Termination date: 20170113 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |