CN101037679A - Nattokinase purification technique and microcapsule preparation technique - Google Patents
Nattokinase purification technique and microcapsule preparation technique Download PDFInfo
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Abstract
The invention relates to a nattokinase microcapsule and preparing method thereof. The nattokinase microcapsule core contains 38-60% of frozen powder, 10-25% of cross-linking CMC-Na, 15-30% of microcrystalline cellulose; the frozen powder is made from good nattokinase which adding addition agent of injection or excipient of conventional mathod. The preparing method of the nattokinase microcapsule is: making frozen powder from good nattokinase which adding addition agent of injection or excipient of conventional mathod; adding frozen powder in alcohol soulution, mixing with cross-linking CMC-Na and microcrystalline cellulose to make softwood, extruding rounded machine-processed nattokinase microcapsule core; drying; adding talcum triethyl citrate by proportion in water, emulsifying at a high speed by emulsification machine to obtain coating liquid, adding and agitating uniformly in EUDRAGIT L30D-55; adding pills in the coating machine of the fluidized bed and spouting coating liquid to coat; coating pills to obtain microcapsule.
Description
Patent application of the present invention is for dividing an application, and the original bill application number is 200410086151.X, and the applying date is on October 27th, 2004, and denomination of invention is natto kinase purifying process and microcapsule formulation technology.
Technical field
The present invention relates to a kind of purifying process and microcapsule and microcapsule formulation technology of biochemical drug, particularly relate to a kind of natto kinase purifying process and enteric-coated microcapsule thereof and enteric-coated microcapsule preparation process.
Background technology
Natto (natto) and Nattokinase (Nattokinase) obtain generally paying attention to both at home and abroad as the research and development of modern food and functional food.Natto is the traditional food of Japan, eats to have exceeded thousand.Nattokinase sees that from palpus in 1987 doctor of foreign firm finds that so far more people pay close attention to, and the focus of concern is that edible Nattokinase food and people's good health and a long life are closely related.It is reported that the Nattokinase goods have very strong solution fibrin effect, blood viscosity lowering, reducing blood-fat, decreasing cholesterol improve the blood circulation situation, keep different physiological roles such as the normal morphology of hemocyte and function.Therefore the Nattokinase preparation is mainly used in prevention and treats heart and brain thrombus, cerebral apoplexy, senile dementia etc.According to the statistical figure that WHO announces, the whole world suffers from the patient of various thrombotic diseases now 1,500 ten thousand more than, and 3,000,000 deaths are wherein arranged every year approximately, finds also that simultaneously this situation is to low age development.Exploitation Nattokinase product has profound significance as prevention and treatment thrombotic diseases, senile dementia etc.
At present the purifying process of Nattokinase adopts following steps: hydrophobic chromatography (filler is Butyl-Toyopearl), ion-exchange (filler is CM-Toyopearl), gel-filtration (SephadexG-50) and dialysis, centrifugal, molten contract etc. just can obtain the elaboration (SDS-PAGE is the wall scroll band) of Nattokinase, it is various that shortcoming is that it produces step, is unfavorable for amplifying, producing.
Nattokinase is a kind of single chain polypeptide enzyme.Because the difference of determination techniques, the molecular weight that records is not waited by 27300KD to 35000KD.Calculating accurate molecular weight according to the aminoacid sequence that has recorded is 27728KD.The iso-electric point of Nattokinase is about 8.6 ± 0.3.To studies show that of substrate specificity, the most responsive substrate of Nattokinase is Suc-Ala-Ala-Pro-Phe-PNA, is H-D-Val-Leu-Lys-PNA (S2251) secondly, and substrate S2238, S2266 and S2302 are had certain activity.Nattokinase is more stable under natural pH state; PH rises at 12 o'clock from 7, and is stable in the 10min; PH is lower than at 5 o'clock, rapidly the sex change inactivation.
Summary of the invention
The object of the invention is to provide a kind of natto kinase purifying process; The object of the invention also is to provide a kind of Nattokinase enteric-coated microcapsule and preparation process thereof.
Natto kinase purifying process of the present invention is achieved by the following technical solution:
Prepare bacillus natto to ferment liquid (as described in No. 02116667.6 patent application) or commercially available Nattokinase crude product according to a conventional method and make solution according to a conventional method; 3000-6000 rev/min centrifugal 15-30 minute; Get supernatant, adding sulphur ammonium to saturation ratio is 30-50%; 3000-6000 rev/min centrifugal; Get precipitation, with the phosphate buffered saline buffer dissolving that contains 0.8-1.2 mol ammonium sulfate of 5 times of precipitation volume amounts, buffer solution ph 6.0-10.0,0.05 mol; Room temperature was placed 12-24 hour; 3000-6000 rev/min centrifugal 15-30 minute; Get supernatant, last hydrophobic chromatography post, filler are phenyl class hydrophobic chromatoghaphy medium; To contain 0.4-1.0 mol ammonium sulfate phosphate buffered saline buffer, buffer solution ph 6.0-10.0,0.05 mol; Gradually to the zero gradient wash-out, the time is about 3-6 hour to the sulphur ammonium concentration with the 0.4-1.0 mol, collects active climax; Ultrafiltration gets the Nattokinase elaboration.
Wherein said Nattokinase crude product liquid can be made as follows: get 100 milligrams of Nattokinase crude products, be dissolved in 20-40 milliliter pH6-9, the phosphate buffered saline buffer of 0.01-0.05 mol, 30-50 ℃ water-bath 8-12 hour;
The present invention also provides a kind of nattokinase microcapsule, contains lyophilized powder 38-60% in the micro-capsule ball core in this microcapsule, crosslinked CMC-Na 10-25%, Microcrystalline Cellulose 15-30%; Described lyophilized powder is to make lyophilized powder by additives or excipient that the Nattokinase elaboration adds injection according to a conventional method.
Preferred lyophilized powder 50% in the micro-capsule ball core in the above-mentioned nattokinase microcapsule, crosslinked CMC-Na25%, Microcrystalline Cellulose 25%.
Microcapsule formulation process program of the present invention is achieved by the following technical solution:
Prepare the Nattokinase elaboration by the inventive method or other ordinary method, the Nattokinase elaboration adds the additives or the excipient (as Dextran 40) of injection according to a conventional method and makes lyophilized powder; It is the 20%-40% ethanolic soln that lyophilized powder adds concentration, the weight ratio of lyophilized powder and ethanolic soln is 1: 0.8-1.2, press lyophilized powder 38-60% again, crosslinked CMC-Na 10-25%, Microcrystalline Cellulose 15-30% adds crosslinked CMC-Na and Microcrystalline Cellulose, mix, the system softwood is extruded round as a ball machine-processed micro-capsule ball core; 30 ℃ of oven dry; Press EUDRAGIT L30D-55 (acrylic resin polymkeric substance) 40%, triethyl citrate 1%, the ratio of water 59%, add the talcum powder triethyl citrate in water, dispersing emulsification machine breast is at a high speed spared 30 minutes, gets coating liquid, its stirring is joined among the EUDRAGIT L30D-55, and stirring causes evenly; Piller is poured in the fluidized-bed coating machine, sprayed into the coating liquid dressing; The dressing parameter is: fluidized-bed temperature 25-28 ℃, and spray pressure 0.2Mpa, air intake flow 0.4-0.45m
3/ min, coating liquid flow velocity 2ml/min; 30 ℃ of baking oven film healings of piller are 8 hours behind the dressing; Coated pellets is encapsulated.
The purifying process of Nattokinase of the present invention adopts primary column chromatography, is the filled column chromatography with Phenyl Sepharose 6Fast flow, the productive rate height, and speed is fast, but ambient operation, condition is easy to control, and it is convenient to handle.The present invention finds that also Nattokinase is relatively stable, thereby the purifying of Nattokinase crude product is greatly simplified.In fact, from the supernatant of the centrifugal back of fermented liquid gained, by alcohol precipitation (getting crude product), thermal destruction (or saltouing), these main three steps of chromatography, finished purifying basically, technology is greatly simplified, the product purity height, cost is low, and yield is higher, can be mass-produced.Nattokinase elaboration and lyophilized powder SDS-PAGE demonstration are the wall scroll band, and molecular weight is 28,000-30,000Dalton; It is 1020 urokinase units/milligram that the EGCT method is measured activity; It is 105 mcg/ml that the Lowry method is measured protein content; Clarity and particulate matter are up to specification; Bacterial endotoxin inspection and aseptic experiment are qualified.Preserved 6 months for 4 ℃-10 ℃, repetition measurement SDS-PAGE still is the wall scroll band, and molecular weight is 28,000-30,000Dalton; It is 1008 urokinase units/milligram that the EGCT method is measured activity, and EGCT method (CLT) shows that elaboration improves more than 10 times than the supernatant liquor specific activity of crude product; It is 103 mcg/ml that the Lowry method is measured protein content; Clarity and particulate matter are up to specification.
The enteric-coated microcapsule preparation technique is adopted in invention, helps reducing loss of activity, and the microcapsule technology preparation granules is little, good evenness, and loss of activity is little, bioavailability height in the body.
Following experimental example is used for explanation but is not limited to the present invention.
Experimental example 1 phenyl hydrophobic chromatography filler (Phenyl--) compares with octyl group hydrophobic chromatography filler (Octyl-)
1. the comparison of applied sample amount
1.1 filler: Phenyl--; Column volume: 2.6 * 30cm; Flow velocity: 5.8cm/min
Table one: the phenyl filler detects
1.2 filler: Octyl--; Column volume: 2.6 * 30cm; Flow velocity: 5.8cm/min
Table two: the octyl group filler detects
Have above two the table as can be known: the applied sample amount of phenyl filler (Phenyl--) more greatly, promptly each quantity of sample handling is than octyl group (Octyl--) about 20%.
2. the comparison of flow velocity
2.1 filler: Phenyl--; Column volume: 2.6 * 30cm; Applied sample amount: 5ml
Table three: the phenyl filler detects
2.2 filler: Octyl--; Column volume: 2.6 * 30cm; Applied sample amount: 5ml
Table four: the octyl group filler detects
By table three, four as can be known, and the peak flow rate of phenyl filler is much larger than the peak flow rate of octyl group filler; Under identical separation condition, the phenyl filler separates the production cycle of Nattokinase much smaller than the octyl group filler.
In sum, the applied sample amount of phenyl filler separation Nattokinase and flow velocity are all greater than the octyl group filler.Use phenyl filler separation natto base enzyme quantity of sample handling bigger, with short production cycle in the production.
Experimental example 2 micro-capsules detect index: (annotate: " 2, release inspection " this a part of article has in full at last)
1, outward appearance: yellow-white piller, spherical rounding, good fluidity, piller particle diameter 400-700 μ m.
2, release inspection: stripping measuring method: little agar diffusion method, rotating speed: 100rpm, temperature: 37 ℃, dissolution medium: among the 0.1mol/L HCl 150ml 2 hours, sampling; In stripping rotor, add 0.2mol/L sodium radio-phosphate,P-32 solution 50ml, continue stripping 45 minutes, sampling.The degassing was handled before dissolution medium used.
3, the control sample enzyme is lived: get three of appeal 6 batch samples respectively, remove the impregnation softgel shell, accurately weigh, put in the 250ml beaker, (0.2mol/L PH6.8 was in 37 ℃ of water-baths 45 minutes to add the 100mlPBS damping fluid, add magnetic agitation, it is fully dissolved, and centrifugal again (4000rpm20min) gets supernatant mensuration enzyme and lives.
Result: all do not measure enzyme in (1) the 2 hour 0.1mol/l hydrochloric acid release liquid and live; (2) 45 minutes PH6.8PBS discharge enzyme and live and control sample enzyme activity determination result, and the average release that the microcapsule formulation enzyme is lived is 96%.
Experimental example 3 micro-capsules detect index:
The inspection of release: stripping measuring method: little agar diffusion method, rotating speed: 100rpm, temperature: 37 ℃, dissolution medium: among the 0.1mol/L hydrochloric acid 150ml 2 hours, sampling; Add 0.2mol/L sodium radio-phosphate,P-32 solution 50ml in stripping rotor, continue stripping 45 minutes, the sampling and measuring enzyme is lived.The degassing was handled before dissolution medium used.
The control sample enzyme is lived: accurately take by weighing above-mentioned microencapsulated sample, put in the 250ml beaker, add 100mlPBS damping fluid (0.2mol/L PH6.8), in 37 ℃ of water-baths 45 minutes, add magnetic agitation, it is fully dissolved, centrifugal again (4000rpm20min) gets supernatant liquor mensuration enzyme and lives.
Result: all do not measure enzyme in (1) the 2 hour 0.1mol/l hydrochloric acid release liquid and live; (2) 45 minutes PH6.8PBS discharge enzyme and live and control sample enzyme activity determination result, and the average release that the microcapsule formulation enzyme is lived is 96%.
Embodiment:
Embodiment 1:
500 milliliters of Nattokinase fermented liquids, 4000 rev/mins are centrifugal 20 minutes; Get supernatant, adding sulphur ammonium to saturation ratio is 40%; 4000 rev/mins centrifugal; Get precipitation, be dissolved in the phosphate buffered saline buffer of 50ml 1.0 mol ammonium sulfate, PH7.0,0.05 mol; Room temperature is placed and is spent the night; 2000 rev/mins centrifugal 15 minutes; Get supernatant, last hydrophobic chromatography post; Filler is a phenyl sepharose, column dimension: 1.6 centimetres of internal diameters, high 20 centimetres, use PH6.0-10.0 in advance, the 0.01-0.05 mol contain 0.8 mol ammonium sulfate phosphate buffered saline buffer balance; Applied sample amount is 30ml, flow velocity 8ml/min, gradient elution; Be that ammonium sulfate concentrations is extremely zero gradually by 0.8 mol, the time is about 4 hours;
The protein nucleic acid ultraviolet detection is beginning that big peak, a non-activity of passing through is arranged; Near 40% o'clock, occurred an active climax in ammonium sulfate concentrations, collected this peak, the ultrafiltration desalination gets 40 milliliters of Nattokinase elaboration, and yield is 15% (elaboration protein milligram/crude product milligram * 100%);
The Nattokinase elaboration is the wall scroll band through SDS-PAGE (Coomassie brilliant blue R250 dyeing and argentation), and molecular weight is 28,000-30,000Dalton; It is 1500 urokinase units/milligram that the EGCT method is measured activity; It is 117.8 mcg/ml that Lowry method (micromethod) is surveyed protein content; Specific activity is 12733 urokinase units/milligram protein.
Embodiment 2:
Get 100 milligrams of Nattokinase crude products, be dissolved in 30 milliliters of PH7.4, the phosphate buffered saline buffer of 0.02 mol; 40 ℃ of water-baths 10 hours; 5000 rev/mins centrifugal 20 minutes; Get and reset and add ammonium sulfate, make whole solubility reach 1.0 mol; 4000 rev/mins centrifugal 20 minutes; Get supernatant, last hydrophobic chromatography post, filler are phenyl sepharose, column dimension: 1.6 centimetres of internal diameters, high 20 centimetres, give and use earlier PH7.0,0.05 mol contain 0.8 mol ammonium sulfate phosphate buffered saline buffer balance; Applied sample amount is 10ml, flow velocity 8ml/min, and gradient elution, promptly ammonium sulfate concentrations is extremely zero gradually by 0.8 mol, about 3 hours of elution time; The albumen ultraviolet detection begins to have big peak, a non-activity of passing through; Near 40% o'clock, occurred an active climax in ammonium sulfate concentrations, collected this peak, the ultra-filtration desalination gets 25 milliliters of Nattokinase elaboration, and yield is 20% (elaboration protein milligram/crude product milligram * 100%);
The Nattokinase elaboration is the wall scroll band through SDS-PAGE (Coomassie brilliant blue R250 dyeing and argentation), and molecular weight is 28,000-30,000Dalton; It is 1200 urokinase units/milligram that the EGCT method is measured activity; It is 93.57 mcg/ml that Lowry method (micromethod) is surveyed protein content; Specific activity is 12824 urokinase units/milligram protein.
Embodiment 3:
It is 1000 urokinase units per ml that the Nattokinase elaboration is regulated concentration, adds 3%-10% Dextran 20-80; Be packed as 1.25 milliliters/cillin bottle, 30 handkerchiefs, minimum temperature is-40 ℃ of lyophilizes 24 hours; Tamponade promptly gets the injection lyophilized powder of Nattokinase;
The lyophilized powder of getting the Nattokinase elaboration is that 1: 1 adding concentration is 30% ethanolic soln by weight, presses lyophilized powder: crosslinked CMC-Na: Microcrystalline Cellulose system softwood, extrude round as a ball machine-processed micro-capsule ball core, 30 ℃ of oven dry; Press EUDRAGIT L30D-55 40%, triethyl citrate 1%, the ratio of water 59% adds triethyl citrate in water, and dispersing emulsification machine breast is at a high speed spared 30 minutes, gets coating liquid; Its stirring is joined among the EUDRAGIT L30D-55, and stirring causes evenly; Piller is poured in the fluidized-bed coating machine, sprayed into the coating liquid dressing; The dressing parameter is: fluidized-bed temperature 25-28 ℃, and spray pressure 0.2Mpa, air intake flow 0.4-0.45m
3/ min, coating liquid flow velocity 2ml/min; 30 ℃ of baking oven film healings of piller are 8 hours behind the dressing; Coated pellets is encapsulated.
The outward appearance yellow-white piller of Nattokinase elaboration enteric-coated microcapsule, spherical rounding, good fluidity, piller particle diameter 400-700 μ m.All not having enzyme in 2 hours 0.1mol/l hydrochloric acid release liquid lives.Enzyme release alive is about 96% in 45 minutes PH6.8PBS damping fluids.
Claims (9)
1, a kind of nattokinase microcapsule is characterized in that containing lyophilized powder 38-60% in the micro-capsule ball core in this microcapsule, crosslinked CMC-Na 10-25%, Microcrystalline Cellulose 15-30%; Described lyophilized powder is made lyophilized powder by additives or excipient that the Nattokinase elaboration adds injection according to a conventional method.
2, nattokinase microcapsule as claimed in claim 1 is characterized in that containing in the micro-capsule ball core in this microcapsule: lyophilized powder 50%, crosslinked CMC-Na 25%, Microcrystalline Cellulose 25%.
3, nattokinase microcapsule as claimed in claim 1 or 2 is characterized in that the Nattokinase elaboration in this microcapsule is prepared by following method:
Get and prepare the solution that bacillus natto to ferment liquid or commercially available Nattokinase crude product are made according to a conventional method according to a conventional method, 3000-6000 rev/min centrifugal 15-30 minute; Get supernatant, adding sulphur ammonium to saturation ratio is 30-50%; 3000-6000 rev/min centrifugal; Get precipitation, with the phosphate buffered saline buffer dissolving that contains 0.8-1.2 mol ammonium sulfate of 5 times of precipitation volume amounts, buffer solution ph 6.0-10.0,0.05 mol; Room temperature was placed 12-24 hour; 3000-6000 rev/min centrifugal 15-30 minute; Get supernatant, last hydrophobic chromatography post, filler are phenyl class hydrophobic chromatoghaphy medium; Use PH6.0-10.0 in advance, the 0.01-0.05 mol contain 0.4-1.0 mol ammonium sulfate phosphate buffered saline buffer balance; Gradually to the zero gradient wash-out, the time is 3-6 hour to the sulphur ammonium concentration with the 0.4-1.0 mol, collects active climax; Ultrafiltration gets the Nattokinase elaboration.
4, nattokinase microcapsule as claimed in claim 3 is characterized in that the Nattokinase elaboration in this microcapsule is prepared by following method: get 500 milliliters of Nattokinase fermented liquids, 4000 rev/mins centrifugal 20 minutes; Get supernatant, adding sulphur ammonium to saturation ratio is 40%; 4000 rev/mins centrifugal; Get precipitation, be dissolved in the phosphate buffered saline buffer of 50ml 1.0 mol ammonium sulfate, PH7.0,0.05 mol; Room temperature is placed and is spent the night; 2000 rev/mins centrifugal 15 minutes; Get supernatant, last hydrophobic chromatography post; Filler is a phenyl sepharose, column dimension: 1.6 centimetres of internal diameters, high 20 centimetres, use PH6.0-10.0 in advance, the 0.01-0.05 mol contain 0.8 mol ammonium sulfate phosphate buffered saline buffer balance; Applied sample amount is 30ml, flow velocity 8ml/min, gradient elution; Be that ammonium sulfate concentrations is extremely zero gradually by 0.8 mol, the time is 4 hours; Collect active climax; Ultrafiltration gets the Nattokinase elaboration.
5, nattokinase microcapsule as claimed in claim 3, it is characterized in that the Nattokinase elaboration in this microcapsule is prepared by following method: get 100 milligrams of Nattokinase crude products, ordinary method is made solution, and 5000 rev/mins are centrifugal 20 minutes; Get and reset and add ammonium sulfate, make whole solubility reach 1.0 mol; 4000 rev/mins centrifugal 20 minutes; Get supernatant, last hydrophobic chromatography post, filler are phenyl sepharose, column dimension: 1.6 centimetres of internal diameters, high 20 centimetres, give and use earlier PH7.0,0.05 mol contain 0.8 mol ammonium sulfate phosphate buffered saline buffer balance; Applied sample amount is 10ml, flow velocity 8ml/min, and gradient elution, promptly ammonium sulfate concentrations is extremely zero gradually by 0.8 mol, elution time 3 hours; The albumen ultraviolet detection begins to have big peak, a non-activity of passing through; Near 40% o'clock, occurred an active climax in ammonium sulfate concentrations, collected this peak, the ultra-filtration desalination gets 25 milliliters of Nattokinase elaboration.
6, nattokinase microcapsule as claimed in claim 5 is characterized in that the Nattokinase crude product solution is made by following method among the Nattokinase elaboration preparation method in this microcapsule:
Get 100 milligrams of Nattokinase crude products, be dissolved in 20-40 milliliter pH6-9, the phosphate buffered saline buffer of 0.01-0.05 mol, 30-50 ℃ water-bath 8-12 hour.
7,, it is characterized in that this method is as the preparation method of claim 1,2,4,5 or 6 described nattokinase microcapsule:
The Nattokinase elaboration adds the additives or the excipient of injection according to a conventional method and makes lyophilized powder; It is the 20%-40% ethanolic soln that lyophilized powder adds concentration, and the weight ratio of lyophilized powder and ethanolic soln is 1: 0.8-1.2, and add crosslinked CMC-Na and Microcrystalline Cellulose again and mix, the system softwood is extruded round as a ball machine-processed micro-capsule ball core; 30 ℃ of oven dry; Press acrylic resin polymkeric substance 40%, triethyl citrate 1%, the ratio of water 59% adds the talcum powder triethyl citrate in water, and dispersing emulsification machine breast is at a high speed spared 30 minutes, gets coating liquid, and its stirring is joined among the EUDRAGIT L30D-55, stirs to cause evenly; Piller is poured in the fluidized-bed coating machine, sprayed into the coating liquid dressing; The dressing parameter is: fluidized-bed temperature 25-28 ℃, and spray pressure 0.2Mpa, air intake flow 0.4-0.45m
3/ min, coating liquid flow velocity 2ml/min; 30 ℃ of baking oven film healings of piller are 8 hours behind the dressing; Coated pellets is encapsulated.
8, the preparation method of a kind of nattokinase microcapsule as claimed in claim 7 is characterized in that this method is:
The Nattokinase elaboration adds the additives or the excipient of injection according to a conventional method and makes lyophilized powder; Getting Nattokinase elaboration lyophilized powder is 1: 1 adding 30% ethanolic soln by weight, adds crosslinked CMC-Na and Microcrystalline Cellulose again and mixes, and the system softwood is extruded round as a ball machine-processed micro-capsule ball core, 30 ℃ of oven dry; Press EUDRAGITL30D-5540%, triethyl citrate 1%, the ratio of water 59% adds triethyl citrate in water, and dispersing emulsification machine breast is at a high speed spared 30 minutes, gets coating liquid; Its stirring is joined among the EUDRAGIT L30D-55, and stirring causes evenly; Piller is poured in the fluidized-bed coating machine, sprayed into the coating liquid dressing; The dressing parameter is: fluidized-bed temperature 25-28 ℃, and spray pressure 0.2Mpa, air intake flow 0.4-0.45m
3/ min, coating liquid flow velocity 2ml/min; 30 ℃ of baking oven film healings of piller are 8 hours behind the dressing; Coated pellets is encapsulated.
9, the preparation method of a kind of nattokinase microcapsule as claimed in claim 7 is characterized in that this method is:
It is 1000 urokinase units per ml that the Nattokinase elaboration is regulated concentration, adds 3%-10% Dextran 20-80; Be packed as 1.25 milliliters/cillin bottle, 30 handkerchiefs, minimum temperature is-40 ℃ of lyophilizes 24 hours; Tamponade promptly gets the injection lyophilized powder of Nattokinase;
Getting Nattokinase elaboration lyophilized powder is 1: 1 adding 30% ethanolic soln by weight, presses lyophilized powder 50% again, crosslinked CMC-Na 25%, Microcrystalline Cellulose 25% adds crosslinked CMC-Na and Microcrystalline Cellulose, the system softwood is extruded round as a ball machine-processed micro-capsule ball core, 30 ℃ of oven dry; Press EUDRAGIT L30D-55 40%, triethyl citrate 1%, the ratio of water 59% adds triethyl citrate in water, and dispersing emulsification machine breast is at a high speed spared 30 minutes, gets coating liquid; Its stirring is joined among the EUDRAGIT L30D-55, and stirring causes evenly; Piller is poured in the fluidized-bed coating machine, sprayed into the coating liquid dressing; The dressing parameter is: fluidized-bed temperature 25-28 ℃, and spray pressure 0.2Mpa, air intake flow 0.4-0.45m
3/ min, coating liquid flow velocity 2ml/min; 30 ℃ of baking oven film healings of piller are 8 hours behind the dressing; Coated pellets is encapsulated.
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| CN102319181A (en) * | 2011-09-08 | 2012-01-18 | 广东大华农动物保健品股份有限公司 | Enveloping process for micro-capsule animal medicament |
| CN103238829A (en) * | 2012-02-13 | 2013-08-14 | 中国科学院过程工程研究所 | Nattokinase enteric capsule and preparation method thereof |
| CN105725128A (en) * | 2016-02-29 | 2016-07-06 | 南通香佳纺织科技有限公司 | Preparation method of microcrystalline cellulose microcapsules by taking konjaku flour as wall materials |
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| CN109646421A (en) * | 2019-01-25 | 2019-04-19 | 纽斯葆广赛（广东）生物科技股份有限公司 | A kind of preparation method of Nattokinase micro-capsule |
| CN110272885A (en) * | 2019-07-05 | 2019-09-24 | 王跃驹 | Plant source Nattokinase capsule and its production method |
| CN112402624A (en) * | 2020-11-13 | 2021-02-26 | 江南大学 | Nattokinase sustained-release microsphere and preparation method thereof |
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2004
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102319181A (en) * | 2011-09-08 | 2012-01-18 | 广东大华农动物保健品股份有限公司 | Enveloping process for micro-capsule animal medicament |
| CN103238829A (en) * | 2012-02-13 | 2013-08-14 | 中国科学院过程工程研究所 | Nattokinase enteric capsule and preparation method thereof |
| CN105725128A (en) * | 2016-02-29 | 2016-07-06 | 南通香佳纺织科技有限公司 | Preparation method of microcrystalline cellulose microcapsules by taking konjaku flour as wall materials |
| CN108771248A (en) * | 2018-05-17 | 2018-11-09 | 吉林修正健康股份有限公司 | A kind of health-care food for assisting blood fat lowering full of nutrition and preparation method thereof |
| CN109646421A (en) * | 2019-01-25 | 2019-04-19 | 纽斯葆广赛（广东）生物科技股份有限公司 | A kind of preparation method of Nattokinase micro-capsule |
| CN110272885A (en) * | 2019-07-05 | 2019-09-24 | 王跃驹 | Plant source Nattokinase capsule and its production method |
| CN112402624A (en) * | 2020-11-13 | 2021-02-26 | 江南大学 | Nattokinase sustained-release microsphere and preparation method thereof |
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