CN104351466A - Low-ash porcine hemoglobin decolorization method - Google Patents
Low-ash porcine hemoglobin decolorization method Download PDFInfo
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- CN104351466A CN104351466A CN201410547869.8A CN201410547869A CN104351466A CN 104351466 A CN104351466 A CN 104351466A CN 201410547869 A CN201410547869 A CN 201410547869A CN 104351466 A CN104351466 A CN 104351466A
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Abstract
The invention relates to a pig blood decolorization method, in particular to a low-ash porcine hemoglobin decolorization method. The method comprises the following steps: preparation of a porcine hemoglobin solution : disodium citrate, sodium bicarbonate and plant phenol extracted from natural plants are mixed in the mass ratio of 1: (2-5): (0.01-0.03) and added into the pig blood, a first supernatant layer and a first sedimentation layer are obtained through centrifugal separation, a first sediment is obtained through separation, and primary ultrasonication is performed; pretreatment: physical deoxidation, pH value regulation and secondary ultrasonication are performed; enzymatic hydrolysis decolorization: an enzyme is added, an enzymatic hydrolysis reaction is ended after enzymatic hydrolysis is performed for 1-2 h, and an enzymatic hydrolysis decolorization solution is obtained; aftertreatment: pH value is regulated, a second supernatant layer and a second sedimentation layer are obtained through centrifugal separation, and the second supernatant layer is subjected to ultrafiltration and concentration to obtain a second concentrated solution; and spray drying: the second concentrated solution is subjected to spray drying to obtain milky porcine hemoglobin polypeptide powder. The method has good decolorization effect and can prepare porcine hemoglobin powder with high protein content and low ash content.
Description
Technical field
The present invention relates to a boar blood discoloration method, particularly relate to a kind of low-ash PINPROL discoloration method.
Background technology
In pig blood, erythrocytic protein content is up to 38%, accounts for more than 75% of whole blood Tot Prot, and wherein more than 92% is hemoglobin, and therefore pig blood cell is a kind of excellent animal protein source, and pig blood processor is directly made into the sale of blood cell powder for a long time.But owing to containing ferroheme in blood cell powder, make blood cell powder be dark red, and with the very heavy smell of blood, accept, and its economic value added is not high sense organ bad being difficult to.
Ferroheme to be combined with four globins with the form of PORPHYRIN IRON to form hemoglobin, is present in red blood cell.In the food industry, the colour former nitrate in the alternative meat products of ferroheme and synthetic food color, use ferroheme can reduce the carcinogenesis of nitrite.In pharmaceuticals industry, ferroheme can be used as semi-synthetic bilirubin raw material, and is anticancer specific drug.Ferroheme, clinically as iron supplementary, can treat the anemia caused by iron deficiency, and this ferroheme iron supplementary can directly be absorbed by the body, and absorptivity, up to 10% ~ 20%, has without the advantages such as bad reaction such as iron accumulate poisoning and gastrointestinal irritation in body.
CN100581377C (2010-1-20) discloses the preparation method of a kind of Apocytochrome porcine hemoglobin enzymolysis matter porcine hemoglobin zymolyte and heme peptide; The method take pig erythrocyte as raw material, by haemolysis, compound protease enzymolysis, etc. electric centrifugation prepare Apocytochrome porcine hemoglobin enzymolysis matter porcine hemoglobin zymolyte and heme peptide.But the ash content that the method prepares product is comparatively large, and decolorizing effect still haves much room for improvement.
Summary of the invention
The object of this invention is to provide a kind of good decolorizing effect, can prepare protein content high, the low-ash PINPROL discoloration method that can improve animal palatability, improve the PINPROL powder of proteopepsis absorptivity and sucking pig immunity.
Above-mentioned technical purpose of the present invention is achieved by the following technical programs:
A kind of low-ash PINPROL discoloration method, it is prepared from by following steps successively:
(1) porcine hemoglobin solution preparation: after the live pig be up to the standards is taken a blood sample, add the disodium citrate mixed according to mass ratio 1:2-5:0.01-0.03, sodium acid carbonate and the plant phenols extracted from natural plants, wherein the weight ratio of disodium citrate and pig blood is 1-2:1000, at 15-25 DEG C of first time disk centrifugal separation 3-6min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, deionized water is added in the first sediment, under the ultrasonic power of 1800-2500W, first time ultrasonic wave process 30-60s obtains porcine hemoglobin solution,
The described plant phenols extracted from natural plants is Tea Polyphenols or apple polyphenol;
(2) pre-treatment: use nitrogen bubble deoxidation to carry out first time physics deoxidation and preliminary decolouring to described porcine hemoglobin solution, then by the described porcine hemoglobin solution adjust ph after deoxidation to 6-7, then second time ultrasonic wave process 50-90s under the ultrasonic power of 600-800W;
(3) enzymolysis decolouring: add the animal proteolytic enzyme of 1:1-2:0.5-0.8:0.2-0.4 mixing in mass ratio, nagarse, papain and pancreatin, the concentration ratio of described animal proteolytic enzyme and described porcine hemoglobin solution is 10000-20000U/g, and after enzymolysis 1-2h, enzymolysis reaction obtains enzymolysis destainer;
(4) post processing: by described enzymolysis destainer adjust ph to 4-5, water-bath 15-25min, is separated 2-5min at 15-25 DEG C of second time disk centrifugal, obtains the second supernatant layer and second beds of precipitation; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.5-0.8MPa and obtain the second concentrate;
Described milipore filter is polyvinylidene fluoride (PVDF) ultrafiltration membrane, and molecular cut off is 6000-8000Da;
(5) spraying dry: described second concentrate is carried out spraying dry and obtains milky PINPROL polypeptide powder.
The disodium citrate that porcine hemoglobin solution of the present invention adds when preparing, sodium acid carbonate and the plant phenols mixture extracted from natural plants can make the anti-freezing of pig blood and anti-sex change, improve its heat endurance, ensure that nutritional labeling does not run off; Carrying out ultrasonic wave process after obtaining sediment is to carry out precrushing to the pig blood cell in sediment, being beneficial to follow-up decolouring; Decolouring adopts that pre-treatment is tentatively decoloured, enzymolysis secondary decolourization and the multistage desolventing technology of post processing and purifying technique, fully can remove color wherein and the smell of blood, make final obtained hemoglobin polypeptide powder color close to milky, protein content is up to more than 95%, and ash content is low to moderate less than 1%; Solubility >=98%, VBN < 130mg/kg; Salmonella, CSFV, aftosa and PCV-II all do not detect.
The invention enables hemoglobin polypeptide powder to be stripped of color and the smell of blood, close to milky, and have frankincense taste, its sensory properties significantly promotes; Hemoglobin polypeptide powder content of ashes is low, and solubility is good, and protein content is high, is especially rich in a large amount of micromolecule polypeptides, and improves the content of several amino acids.The feed nutrition of processing has improvement, can improve sucking pig immunity, increase antibacterial and to virus inhibitory action.
As preferably, get the second beds of precipitation material obtained in described post-processing step, at-10--15 DEG C of freezing 1-2h, naturally add 1-2 times of deionized water washing after thawing, then carry out centrifugation, get the sediment that centrifugation obtains, after adding 0.1-0.2 times of deionized water washing, carry out the deoxidation of second time physics, then centrifugation again, last freeze drying obtains heme peptide.
With original pig blood for raw material, multistep treatment technology and the method is coordinated to obtain ferroheme crude product accessory substance through biological enzymolysis.Adopt the ferroheme rate of recovery prepared of the inventive method to reach more than 95%, the content of hemachrome in heme peptide reaches 40%-45%, and color is aubergine, and without fishy smell, inventor finds, this is with to carry out again the deoxidation of second time physics after freezing separation relevant.
As preferably, described spray-dired EAT is 190-240 DEG C, and leaving air temp is 60-70 DEG C, and maximum water evaporation quantity is 3-5kg/h.
As preferably, after the live pig be up to the standards specifically is taken a blood sample by described porcine hemoglobin solution preparation, add the disodium citrate mixed according to mass ratio 1:3:0.03, sodium acid carbonate and the plant phenols extracted from natural plants, wherein the weight ratio of disodium citrate and pig blood is 1.5:1000, at 20 DEG C of first time disk centrifugal separation 4min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, the deionized water of 1.8-2 times of volume is added in the first sediment, under the ultrasonic power of 2000W, first time ultrasonic wave process 50s obtains porcine hemoglobin solution.
As preferably, described post processing is by described enzymolysis destainer adjust ph to 4-5, is process 15-25min in the water-bath of 10-18 DEG C in temperature, is separated 3min, obtains the second supernatant layer and second beds of precipitation at 18 DEG C of second time disk centrifugals; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.6MPa and obtain the second concentrate.
As preferably, the disk centrifuge that described first time dish-style separation adopts, comprise body, rotary drum is provided with in body described in it, described body top is provided with inlet and outlet device, described inlet and outlet device comprises the material inlet pipe be arranged in parallel, heavy out pipe and light phase export pipe, described material inlet pipe, heavy out pipe and light phase export pipe communicate with described rotary drum, described body side is provided with the deslagging portion and wastewater outlet that communicate with described rotary drum, described wastewater outlet is positioned at the below in described deslagging portion, the side of described body is provided with rinse water import, PLC device is provided with in described deslagging portion,
Described inlet and outlet device comprises the material inlet pipe imported and exported seat and be fixed on described import and export seat, the sidewall of described material inlet pipe is provided with the upper centripetal discharge pump for separating of heavy phase and the lower centripetal discharge pump for separating of light phase, described upper centripetal discharge pump is positioned at the top of described lower centripetal discharge pump, the side of described material inlet pipe is connected with in described material inlet caliber one end to the heavy out pipe extended, the opposite side of described material inlet pipe is connected with in described material inlet caliber one end to the light phase export pipe extended, the other end of described heavy out pipe and described light phase export pipe is respectively arranged with flow regulator.
Advantage of the present invention is:
(1) programmable logic controller (PLC) (PLC) device is adopted to be arranged on described deslagging portion, regularly quantitative deslagging can be carried out when continuous feed is produced, also total discharge can be carried out when stopping charging, so that the dirt automatically in cleaning rotary drum, have that automaticity is high, advantage to the strong adaptability of technique adjustment, easy to adjust, good separating effect;
(2) by the layout of described material inlet pipe, heavy out pipe and light phase export pipe, coordinate the setting in rotary drum and deslagging portion simultaneously, the heavy burder blood cell that density can be made relatively large is flowed out by lower centripetal pump chamber, the light material blood plasma making density relatively little flows out from lower centripetal pump chamber, and the separating property be separated with blood cell pig blood blood plasma is high, effective;
(3) layout of described material inlet pipe and described upper centripetal discharge pump and lower centripetal discharge pump is crossed, coordinate heavy out pipe, light phase export pipe and flow regulator simultaneously, the heavy burder blood cell that density can be made relatively large is flowed out by lower centripetal discharge pump, the light material blood plasma making density relatively little flows out from lower centripetal discharge pump, and the separating property be separated with blood cell pig blood blood plasma is high, effective.
As preferably, described rotary drum comprises main shaft, the pedestal of fixing described main shaft and the rotary drum body adjacent with described pedestal, it also comprises and is pressed in described rotary drum body and is distributed in video disc gland and the rotary drum lid of described main shaft both sides, it is the feed distribution pipe of 30-60 ° of angle that described main shaft sidewall is installed with described main shaft, the video disc of uneven layout is provided with in described rotary drum body, the angle of described video disc and described main shaft is 30-60 °, described spindle top is provided with the gravity disc for separating of different densities material, the upper centripetal pump chamber for separating of heavy phase is provided with above described gravity disc, the lower centripetal pump chamber for separating of light phase is provided with below described gravity disc.
The present invention is arranged by the angle of described feed distribution pipe and described video disc, coordinate the setting of gravity disc and upper centripetal pump chamber and lower centripetal pump chamber simultaneously, the heavy burder blood cell that density can be made relatively large is flowed out by lower centripetal pump chamber, the light material blood plasma making density relatively little flows out from lower centripetal pump chamber, and the separating property be separated with blood cell pig blood blood plasma is high, effective.
More preferably, the angle of described video disc gland and described main shaft is 30-60 °, and the angle of described rotary drum lid and described main shaft is 30-60 °.
More preferably, below described feed distribution pipe and the adjacent being positioned at described pedestal and described rotary drum body is provided with O RunddichtringO; The circumference of described O RunddichtringO is symmetrically arranged with the twice otch for clamping seal groove, and the line angle in described twice otch and the described O RunddichtringO center of circle is 90 °.
O RunddichtringO can make rotary drum better tightness, thus improves the separating effect of blood plasma and blood cell.
More preferably, described flow regulator comprises the Flow-rate adjustment room communicated with the other end of described heavy out pipe or described light phase export pipe, and described Flow-rate adjustment room is connected with pressure apparatus.
Regulated by the pressure in Flow-rate adjustment room, heavy phase material and the inlet and outlet pressure of gently expecting mutually can be controlled, thus improve separating effect.
In sum, the present invention has following beneficial effect:
1, the PINPROL polypeptide powder that prepared by the present invention is stripped of color and the smell of blood, and close to milky, and have frankincense taste, its sensory properties significantly promotes;
2, the PINPROL content prepared of the present invention high, animal palatability can be improved, improve proteopepsis absorptivity and sucking pig immunity;
3, the present invention also can obtain the ferroheme crude product accessory substance of high added value simultaneously.
Accompanying drawing explanation
Fig. 1 is the front view of disk centrifuge of the present invention;
Fig. 2 is the top view of disk centrifuge of the present invention;
Fig. 3 is the side view of disk centrifuge of the present invention;
Fig. 4 is the operating diagram of disk centrifuge of the present invention;
Fig. 5 is disc type separator rotary drum sectional view of the present invention;
Fig. 6 is O RunddichtringO enlarged diagram of the present invention;
Fig. 7 is the schematic diagram of inlet and outlet device of the present invention;
In figure, 1-rotary drum; 2-inlet and outlet device; 3-deslagging portion; 4-wastewater outlet; The import of 5-rinse water;
11-main shaft; 12-pedestal; 13-rotary drum body; 14-video disc gland; 15-rotary drum lid; 16-feed distribution pipe; 17-video disc; 18-gravity disc; The upper centripetal pump chamber of 19-; Centripetal pump chamber under 110-; The clamp ring of 111-abnormity; 112-locating piece; Hexagonal cone end holding screw in 113-; 114-hydroecium; 115-O RunddichtringO; 116-otch;
21-imports and exports seat; 22-material inlet pipe; The upper centripetal discharge pump of 23-; Centripetal discharge pump under 24-; 25-heavy out pipe; 26-light phase export pipe; 27-flow regulator; 271-Flow-rate adjustment room; 272-pressure apparatus; 273-first endoscopy glass; 28-inlet connector; 281-second endoscopy glass.
Detailed description of the invention
Below in conjunction with accompanying drawing, the present invention is described in further detail.
Embodiment one
Prepared by porcine hemoglobin solution: after being taken a blood sample by the live pig be up to the standards, add disodium citrate, sodium acid carbonate and the Tea Polyphenols or apple polyphenol that mix according to mass ratio 1:5:0.03, wherein the weight ratio of disodium citrate and pig blood is 2:1000, at 25 DEG C of first time disk centrifugal separation 6min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, in the first sediment, add deionized water, under the ultrasonic power of 2500W, first time ultrasonic wave process 60s obtains porcine hemoglobin solution;
Pre-treatment: use nitrogen bubble deoxidation to carry out first time physics deoxidation and preliminary decolouring to porcine hemoglobin solution, then by the porcine hemoglobin solution adjust ph to 7 after deoxidation, then second time ultrasonic wave process 90s under the ultrasonic power of 800W;
Enzymolysis decolours: add the animal proteolytic enzyme of 1:2:0.8:0.4 mixing in mass ratio, nagarse, papain and pancreatin, the concentration ratio of animal proteolytic enzyme and porcine hemoglobin solution is 20000U/g, and after enzymolysis 2h, enzymolysis reaction obtains enzymolysis destainer;
Post processing: by enzymolysis destainer adjust ph to 5, water-bath 25min, is separated 5min at 25 DEG C of second time disk centrifugals, obtains the second supernatant layer and second beds of precipitation; Get the second supernatant layer and be separated acquisition second concentrate by the PVDF ultrafiltration membrane filtering and concentrating that molecular cut off is 8000Da under pressure is 0.8MPa;
Spraying dry: the second concentrate is carried out spraying dry and obtain milky PINPROL polypeptide powder, spray-dired EAT is 240 DEG C, and leaving air temp is 70 DEG C, and maximum water evaporation quantity is 5kg/h.Hemoglobin polypeptide powder color is close to milky, and protein content is up to more than 95%, and ash content is low to moderate less than 1%; Solubility >=98%, VBN < 130mg/kg; Salmonella, CSFV, aftosa and PCV-II all do not detect.
Get the second beds of precipitation material obtained in post-processing step, at-15 DEG C of freezing 2h, naturally 2 times of deionized water washings are added after thawing, then centrifugation is carried out, get the sediment that centrifugation obtains, after adding 0.2 times of deionized water washing, carry out the deoxidation of second time physics, then centrifugation again, last freeze drying obtains heme peptide.The ferroheme rate of recovery reaches more than 95%, and the content of hemachrome in heme peptide reaches 40%, and color is aubergine, and without fishy smell.
As shown in Fig. 1-Fig. 7, the disk centrifuge that first time dish-style separation adopts comprises body, rotary drum 1 is provided with in body, body top is provided with inlet and outlet device 2, inlet and outlet device 2 comprises the material inlet pipe 22, heavy out pipe 25 and the light phase export pipe 26 that are arranged in parallel, material inlet pipe 22, heavy out pipe 25 and light phase export pipe 26 communicate with rotary drum 1, body side is provided with the deslagging portion 3 and wastewater outlet 4 that communicate with rotary drum 1, wastewater outlet 4 is positioned at the below in deslagging portion 3, the side of body is provided with rinse water import 5, is provided with PLC device in deslagging portion 3.Rotary drum 1 comprises main shaft 11, the pedestal 12 of fixed main shaft 11 and the rotary drum body 13 adjacent with pedestal 12, it also comprises and is pressed in rotary drum body 13 and is distributed in video disc gland 14 and the rotary drum lid 15 of main shaft 11 both sides, main shaft 11 sidewall is installed with and the feed distribution pipe 16 of main shaft 11 in 30-60 ° of angle, the video disc 17 of uneven layout is provided with in rotary drum body 13, video disc 17 is 30-60 ° with the angle of main shaft 11, main shaft 11 top is provided with the gravity disc for separating of different densities material, the upper centripetal pump chamber 19 for separating of heavy phase is provided with above gravity disc, the lower centripetal pump chamber 110 for separating of light phase is provided with below gravity disc.Video disc gland 14 is 30-60 ° with the angle of main shaft 11, preferably 50 °; Rotary drum lid 15 is 30-60 ° with the angle of main shaft 11, preferably 50 °.The vertical range of video disc gland 14 and rotary drum lid 15 is 4cm.Rotary drum lid 15 is provided with special-shaped clamp ring 111 and locating piece 112.Be provided with the hydroecium 114 for backwash below pedestal 12, rinse water import 5 communicates with hydroecium 114.Be positioned at pedestal 12 adjacent of rotary drum body 13 is provided with O RunddichtringO 115 below feed distribution pipe 16.The circumference of O RunddichtringO 115 is symmetrically arranged with the twice otch 116 for clamping seal groove, and twice otch 116 is 90 ° with the line angle in O RunddichtringO 115 center of circle.Inlet and outlet device 2 comprises to be imported and exported seat 21 and is fixed on the material inlet pipe 22 imported and exported on seat 21, the sidewall of material inlet pipe 22 is provided with the upper centripetal discharge pump 23 for separating of heavy phase and the lower centripetal discharge pump 24 for separating of light phase, upper centripetal discharge pump 23 is positioned at the top of lower centripetal discharge pump 24, the side of material inlet pipe 22 is connected with the one end of the heavy out pipe 25 extended in material inlet pipe 22 radial direction, the opposite side of material inlet pipe 22 is connected with the one end of the light phase export pipe 26 extended in material inlet pipe 22 radial direction, heavy out pipe 25 is respectively arranged with flow regulator 27 with the other end of light phase export pipe 26.Flow regulator 27 comprises the Flow-rate adjustment room 271 communicated with the other end of heavy out pipe 25 or light phase export pipe 26, and Flow-rate adjustment room 271 is connected with pressure apparatus 272.Rotary drum body 13 is provided with by hexagonal cone end holding screw 113 in further for feed distribution pipe 16 fixing, Flow-rate adjustment room 271 is provided with the first endoscopy glass 273.
Be provided with inlet connector 28 above the top of material inlet pipe, inlet connector 28 be provided with the second endoscopy glass 281.By the installation of the first endoscopy glass and the second endoscopy glass, separation case heavy phase material can observed more accurately and gently expect mutually.
Embodiment two
Prepared by porcine hemoglobin solution: after being taken a blood sample by the live pig be up to the standards, add disodium citrate, sodium acid carbonate and the Tea Polyphenols or apple polyphenol that mix according to mass ratio 1:2:0.01, wherein the weight ratio of disodium citrate and pig blood is 1:1000, at 15 DEG C of first time disk centrifugal separation 3min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, in the first sediment, add deionized water, under the ultrasonic power of 1800W, first time ultrasonic wave process 30s obtains porcine hemoglobin solution;
Pre-treatment: use nitrogen bubble deoxidation to carry out first time physics deoxidation and preliminary decolouring to porcine hemoglobin solution, then by the porcine hemoglobin solution adjust ph to 6 after deoxidation, then second time ultrasonic wave process 50s under the ultrasonic power of 600W;
Enzymolysis decolours: add the animal proteolytic enzyme of 1:1:0.5:0.2 mixing in mass ratio, nagarse, papain and pancreatin, the concentration ratio of animal proteolytic enzyme and porcine hemoglobin solution is 10000U/g, and after enzymolysis 1h, enzymolysis reaction obtains enzymolysis destainer;
Post processing: by enzymolysis destainer adjust ph to 4, water-bath 15min, is separated 2min at 15-25 DEG C of second time disk centrifugal, obtains the second supernatant layer and second beds of precipitation; Get the second supernatant layer and be separated acquisition second concentrate by the PVDF ultrafiltration membrane filtering and concentrating that molecular cut off is 6000Da under pressure is 0.5MPa;
Spraying dry: the second concentrate is carried out spraying dry and obtain milky PINPROL polypeptide powder, spray-dired EAT is 190 DEG C, and leaving air temp is 60 DEG C, and maximum water evaporation quantity is 3kg/h.Hemoglobin polypeptide powder color is close to milky, and protein content is up to more than 95%, and ash content is low to moderate less than 1%; Solubility >=98%, VBN < 130mg/kg; Salmonella, CSFV, aftosa and PCV-II all do not detect.
Get the second beds of precipitation material obtained in post-processing step, at-10 DEG C of freezing 1h, naturally 1-2 times of deionized water washing is added after thawing, then centrifugation is carried out, get the sediment that centrifugation obtains, after adding 0.1 times of deionized water washing, carry out the deoxidation of second time physics, then centrifugation again, last freeze drying obtains heme peptide.The ferroheme rate of recovery reaches more than 95%, and the content of hemachrome in heme peptide reaches 45%, and color is aubergine, and without fishy smell.
Embodiment three
After the live pig be up to the standards specifically is taken a blood sample by the preparation of porcine hemoglobin solution, add the disodium citrate mixed according to mass ratio 1:3:0.03, sodium acid carbonate and the plant phenols extracted from natural plants, wherein the weight ratio of disodium citrate and pig blood is 1.5:1000, at 20 DEG C of first time disk centrifugal separation 4min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, the deionized water of 1.8-2 times of volume is added in the first sediment, under the ultrasonic power of 2000W, first time ultrasonic wave process 50s obtains porcine hemoglobin solution.
Pre-treatment: use nitrogen bubble deoxidation to carry out first time physics deoxidation and preliminary decolouring to porcine hemoglobin solution, then by the porcine hemoglobin solution adjust ph to 6.5 after deoxidation, then second time ultrasonic wave process 70s under the ultrasonic power of 700W;
Enzymolysis decolours: add the animal proteolytic enzyme of 1:1.5:0.6:0.3 mixing in mass ratio, nagarse, papain and pancreatin, the concentration ratio of animal proteolytic enzyme and porcine hemoglobin solution is 16000U/g, and after enzymolysis 1-2h, enzymolysis reaction obtains enzymolysis destainer;
Post processing is by enzymolysis destainer adjust ph to 4, is to process 18min in the water-bath of 15 DEG C in temperature, is separated 3min, obtains the second supernatant layer and second beds of precipitation at 18 DEG C of second time disk centrifugals; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.6MPa and obtain the second concentrate.
Spraying dry: the second concentrate is carried out spraying dry and obtain milky PINPROL polypeptide powder, spray-dired EAT is 200 DEG C, and leaving air temp is 65 DEG C, and maximum water evaporation quantity is 4kg/h.
Hemoglobin polypeptide powder color is close to milky, and protein content is up to more than 95%, and ash content is low to moderate less than 1%; Solubility >=98%, VBN < 130mg/kg; Salmonella, CSFV, aftosa and PCV-II all do not detect.
Get the second beds of precipitation material obtained in post-processing step, at-12 DEG C of freezing 1.5h, naturally 1.5 times of deionized water washings are added after thawing, then centrifugation is carried out, get the sediment that centrifugation obtains, after adding 0.15 times of deionized water washing, carry out the deoxidation of second time physics, then centrifugation again, last freeze drying obtains heme peptide.The ferroheme rate of recovery reaches more than 95%, and the content of hemachrome in heme peptide reaches 45%, and color is aubergine, and without fishy smell.
Comparative example one
With embodiment one, unlike porcine hemoglobin solution preparation be by be up to the standards live pig blood sampling after, add disodium citrate, the weight ratio of disodium citrate and pig blood is 1-2:1000, at 15-25 DEG C of drum-type centrifugation 3-6min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, in the first sediment, add deionized water.Finally obtained hemoglobin polypeptide powder color is canescence, and protein content is 92%, and ash content is 4%.
Comparative example two
With embodiment two, do not carry out pre-treatment unlike after the preparation of porcine hemoglobin solution, but directly carry out enzymolysis decolouring and post processing.
Finally obtained hemoglobin polypeptide powder color is canescence, and protein content is 90%, and ash content is 5%.
Comparative example three
With embodiment three, animal proteolytic enzyme and the nagarse adding 1:3 mixing in mass ratio unlike enzymolysis decolouring, the concentration ratio of animal proteolytic enzyme and porcine hemoglobin solution is 15000U/g, and after enzymolysis 1-2h, enzymolysis reaction obtains enzymolysis destainer; Post processing is by enzymolysis destainer adjust ph to 6-7, water-bath 15min, is separated 5min, obtains the second supernatant layer and second beds of precipitation at 15 DEG C of second time disk centrifugals; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.4MPa and obtain the second concentrate.
Finally obtained hemoglobin polypeptide powder color is canescence, and protein content is 89%, and ash content is 6%.
Comparative example four
With embodiment three, preparing unlike heme peptide is get the second beds of precipitation material obtained in post-processing step, at-10 DEG C of freezing 1-2h, naturally add 1-2 times of deionized water washing after thawing, then carry out centrifugation, get the sediment that centrifugation obtains, after adding 0.1-0.2 times of deionized water washing, then centrifugation again, do not carry out the deoxidation of second time physics, direct freeze drying obtains.The ferroheme rate of recovery is 90%, and the content of hemachrome in heme peptide is 30%, and color is aubergine, slightly fishy smell.
This specific embodiment is only explanation of the invention; it is not limitation of the present invention; those skilled in the art can make to the present embodiment the amendment not having creative contribution as required after reading this description, as long as but in right of the present invention, be all subject to Patent Law be strength protection.
Claims (10)
1. a low-ash PINPROL discoloration method, is characterized in that being prepared from by following steps successively:
(1) porcine hemoglobin solution preparation: after the live pig be up to the standards is taken a blood sample, add the disodium citrate mixed according to mass ratio 1:2-5:0.01-0.03, sodium acid carbonate and the plant phenols extracted from natural plants, wherein the weight ratio of disodium citrate and pig blood is 1-2:1000, at 15-25 DEG C of first time disk centrifugal separation 3-6min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, deionized water is added in the first sediment, under the ultrasonic power of 1800-2500W, first time ultrasonic wave process 30-60s obtains porcine hemoglobin solution,
The described plant phenols extracted from natural plants is Tea Polyphenols or apple polyphenol;
(2) pre-treatment: use nitrogen bubble deoxidation to carry out first time physics deoxidation and preliminary decolouring to described porcine hemoglobin solution, then by the described porcine hemoglobin solution adjust ph after deoxidation to 6-7, then second time ultrasonic wave process 50-90s under the ultrasonic power of 600-800W;
(3) enzymolysis decolouring: add the animal proteolytic enzyme of 1:1-2:0.5-0.8:0.2-0.4 mixing in mass ratio, nagarse, papain and pancreatin, the concentration ratio of described animal proteolytic enzyme and described porcine hemoglobin solution is 10000-20000U/g, and after enzymolysis 1-2h, enzymolysis reaction obtains enzymolysis destainer;
(4) post processing: by described enzymolysis destainer adjust ph to 4-5, water-bath 15-25min, is separated 2-5min at 15-25 DEG C of second time disk centrifugal, obtains the second supernatant layer and second beds of precipitation; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.5-0.8MPa and obtain the second concentrate;
Described milipore filter is polyvinylidene fluoride (PVDF) ultrafiltration membrane, and molecular cut off is 6000-8000Da;
(5) spraying dry: described second concentrate is carried out spraying dry and obtains milky PINPROL polypeptide powder.
2. the low-ash PINPROL discoloration method of one according to claim 1, it is characterized in that: get the second beds of precipitation material obtained in described post-processing step, at-10--15 DEG C of freezing 1-2h, naturally add 1-2 times of deionized water washing after thawing, then carry out centrifugation, get the sediment that centrifugation obtains, after adding 0.1-0.2 times of deionized water washing, carry out the deoxidation of second time physics, then centrifugation again, last freeze drying obtains heme peptide.
3. the low-ash PINPROL discoloration method of one according to claim 2, it is characterized in that: described spray-dired EAT is 190-240 DEG C, leaving air temp is 60-70 DEG C, and maximum water evaporation quantity is 3-5kg/h.
4. the low-ash PINPROL discoloration method of one according to claim 3, it is characterized in that: after the live pig be up to the standards specifically is taken a blood sample by described porcine hemoglobin solution preparation, add the disodium citrate mixed according to mass ratio 1:3:0.03, sodium acid carbonate and the plant phenols extracted from natural plants, wherein the weight ratio of disodium citrate and pig blood is 1.5:1000, at 20 DEG C of first time disk centrifugal separation 4min after mixing, obtain the first supernatant layer and first beds of precipitation, be separated acquisition first sediment, the deionized water of 1.8-2 times of volume is added in the first sediment, under the ultrasonic power of 2000W, first time ultrasonic wave process 50s obtains porcine hemoglobin solution.
5. the low-ash PINPROL discoloration method of one according to claim 4, it is characterized in that: described post processing is to 4-5 by described enzymolysis destainer adjust ph, be process 15-25min in the water-bath of 10-18 DEG C in temperature, be separated 3min at 18 DEG C of second time disk centrifugals, obtain the second supernatant layer and second beds of precipitation; Get the second supernatant layer ultrafiltration membrance filter concentrating and separating under pressure is 0.6MPa and obtain the second concentrate.
6. the low-ash PINPROL discoloration method of the one according to any one of claim 1-5, it is characterized in that: the disk centrifuge that described first time dish-style separation adopts, comprise body, rotary drum (1) is provided with in described body, described body top is provided with inlet and outlet device (2), described inlet and outlet device (2) comprises the material inlet pipe (22) be arranged in parallel, heavy out pipe (25) and light phase export pipe (26), described material inlet pipe (22), heavy out pipe (25) and light phase export pipe (26) communicate with described rotary drum (1), described body side is provided with the deslagging portion (3) and wastewater outlet (4) that communicate with described rotary drum (1), described wastewater outlet (4) is positioned at the below of described deslagging portion (3), the side of described body is provided with rinse water import (5), PLC device is provided with in described deslagging portion (3),
Described inlet and outlet device (2) comprises the material inlet pipe (22) imported and exported seat (21) and be fixed on described import and export seat (21), the sidewall of described material inlet pipe (22) is provided with the upper centripetal discharge pump (23) for separating of heavy phase and the lower centripetal discharge pump (24) for separating of light phase, described upper centripetal discharge pump (23) is positioned at the top of described lower centripetal discharge pump (24), the side of described material inlet pipe (22) is connected with the one end of the heavy out pipe (25) extended in described material inlet pipe (22) radial direction, the opposite side of described material inlet pipe (22) is connected with the one end of the light phase export pipe (26) extended in described material inlet pipe (22) radial direction, described heavy out pipe (25) is respectively arranged with flow regulator (27) with the other end of described light phase export pipe (26).
7. the low-ash PINPROL discoloration method of one according to claim 6, it is characterized in that: described rotary drum (1) comprises main shaft (11), the pedestal (12) of fixing described main shaft (11) and the rotary drum body (13) adjacent with described pedestal (12), it also comprises and is pressed in described rotary drum body (13) and is distributed in video disc gland (14) and the rotary drum lid (15) of described main shaft (11) both sides, described main shaft (11) sidewall is installed with the feed distribution pipe (16) with described main shaft (11) ° angle in 30-60, the video disc (17) of uneven layout is provided with in described rotary drum body (13), described video disc (17) is 30-60 ° with the angle of described main shaft (11), described main shaft (11) top is provided with the gravity disc for separating of different densities material, the upper centripetal pump chamber (19) for separating of heavy phase is provided with above described gravity disc, the lower centripetal pump chamber (110) for separating of light phase is provided with below described gravity disc.
8. the low-ash PINPROL discoloration method of one according to claim 7, it is characterized in that: described video disc gland (14) is 30-60 ° with the angle of described main shaft (11), described rotary drum lid (15) is 30-60 ° with the angle of described main shaft (11).
9. the low-ash PINPROL discoloration method of one according to claim 8, is characterized in that: described feed distribution pipe (16) below and be positioned at described pedestal (12) adjacent of described rotary drum body (13) is provided with O RunddichtringO (115);
The circumference of described O RunddichtringO (115) is symmetrically arranged with the twice otch (116) for clamping seal groove, and described twice otch (116) is 90 ° with the line angle in described O RunddichtringO (115) center of circle.
10. the low-ash PINPROL discoloration method of one according to claim 9, it is characterized in that: described flow regulator (27) comprises the Flow-rate adjustment room (271) communicated with the other end of described heavy out pipe (25) or described light phase export pipe (26), and described Flow-rate adjustment room (271) is connected with pressure apparatus (272).
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