CN109172607B - Process and method for extracting placenta from fresh donkey placenta - Google Patents

Process and method for extracting placenta from fresh donkey placenta Download PDF

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Publication number
CN109172607B
CN109172607B CN201811346112.7A CN201811346112A CN109172607B CN 109172607 B CN109172607 B CN 109172607B CN 201811346112 A CN201811346112 A CN 201811346112A CN 109172607 B CN109172607 B CN 109172607B
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donkey
placenta
deodorization
donkey placenta
fresh
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CN109172607A (en
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潘玉柱
董金豹
何涛
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Hello Donkey Tianjin International Trade Co ltd
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Hello Donkey Tianjin International Trade Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/48Reproductive organs
    • A61K35/50Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K8/00Cosmetics or similar toilet preparations
    • A61K8/18Cosmetics or similar toilet preparations characterised by the composition
    • A61K8/96Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/981Cosmetics or similar toilet preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of mammals or bird
    • A61K8/982Reproductive organs; Embryos, Eggs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00Drugs for immunological or allergic disorders
    • A61P37/02Immunomodulators
    • A61P37/04Immunostimulants
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00General protective or antinoxious agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00General protective or antinoxious agents
    • A61P39/06Free radical scavengers or antioxidants
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILET PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D11/00Solvent extraction
    • B01D11/02Solvent extraction of solids
    • B01D11/0215Solid material in other stationary receptacles
    • B01D11/0253Fluidised bed of solid materials
    • B01D11/0257Fluidised bed of solid materials using mixing mechanisms, e.g. stirrers, jets
    • BPERFORMING OPERATIONS; TRANSPORTING
    • B01PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01DSEPARATION
    • B01D36/00Filter circuits or combinations of filters with other separating devices

Abstract

The invention discloses a process and a method for extracting donkey placentin from a fresh donkey placenta, belonging to the technical field of biology. The invention quarantines the live donkey before the raw material is obtained, directly takes out the fresh donkey placenta tissue in the slaughtering process of the live donkey, the fresh donkey placenta tissue has high biological activity and is easy to extract the placentin, meanwhile, the invention is always carried out under the aseptic condition, which is favorable for maintaining the biological activity of trypsin and micromolecule components in the placenta extract, the method of the invention is provided with a deodorization and filtration integrated device, the fishy smell removing device and the filter box are arranged in the tank body, the supernatant is poured into the fishy smell removing device through the feeding hole, the donkey placenta extract is filtered by the loose cotton and the activated carbon to remove fishy smell and then enters a filter box, a vacuum suction filter pump is used for suction filtration, and supernatant liquid after fishy smell removal can be filtered by a microporous filter membrane to obtain clarified liquid.

Description

Process and method for extracting placenta from fresh donkey placenta
Technical Field
The invention relates to the technical field of biology, in particular to a process and a method for extracting placentin from fresh donkey placenta.
Background
Placentin, colloquially, is divided into two types, namely, animal-derived placentin and human-derived placentin, and in practice, the animal-derived placentin is also called sheep placenta. However, human placental peptide can only be obtained from human placenta. They are not specific names of products, but are just general names of certain types of products. Placental peptide is a commonly used health product, is a small molecular compound with immunological activity extracted from placenta, and is also called placenta peptide. The theory of traditional Chinese medicine considers that the placenta has the functions of tonifying qi, nourishing blood, replenishing vital essence and the like, has health-care effect on skin, has great influence on immunity and motion ability, and has the functions of resisting fatigue, resisting aging, improving cellular immunity and the like.
Due to the many effects of placenta, the extraction and isolation of its active ingredients (polypeptides, SOD, ATP, HASF, EGF, etc.) using modern biotechnology methods and modern chemical methods has become a hot point of research. The method of extracting sheep placenta by dialysis and ultrafiltration and processing the sheep placenta into high-tech products applied to clinical medicine, skin care cosmetics and the like has been reported for a long time. The placenta extract generally contains main components including proteins, nucleic acids, phospholipids, polysaccharides, amino acids, minerals, vitamins and the like, and essence of the placenta extract is small-molecular peptide substances and nucleic acid fragments which are also basic elements for activating cells, so that the placenta extract is extracted while impurities are removed and the activity of effective components is kept. In the prior art, the placenta raw material is usually pretreated by adopting a cooking method, the high temperature can destroy the effective components in the placenta extract, and the deodorization and filtration treatment process of the placenta tissue concentrated solution is complicated and has poor deodorization effect.
Disclosure of Invention
The invention aims to solve the problems that the method for extracting the placentin by pretreating the placenta at high temperature in the prior art can destroy effective components in the placentin, and the fishy smell removing treatment process is simple and the fishy smell removing effect is poor in the prior art, and provides a process and a method for extracting the donkey placentin from fresh donkey placenta.
In order to achieve the purpose, the invention adopts the following technical scheme:
a process and method for extracting donkey placentin from fresh donkey placenta comprise the following steps:
s1, quarantining the live donkeys and then sending the live donkeys into a large slaughter house for slaughtering, after slaughtering the live donkeys, sending the live donkeys into a food-grade production and processing environment, quickly taking down fresh donkey placenta tissues from the abdomen of a mother donkey and cleaning the fresh donkey placenta tissues in the slaughtering process, and then putting the donkey placenta tissues into a vacuum freeze dryer for freeze preservation for later use;
s2, taking out the donkey placenta tissue freeze-dried in S1, and crushing the donkey placenta tissue by using a tissue crusher under the aseptic condition to prepare donkey placenta tissue raw powder;
s3, under the aseptic condition, weighing a proper amount of donkey placenta tissue raw powder in the S2, adding a proper amount of sterilized deionized water into the donkey placenta tissue raw powder according to the mass ratio of 1:1-1:5, and homogenizing by a homogenizer to prepare donkey placenta tissue homogenate;
s4, opening the high-speed centrifuge for precooling, placing the donkey fetal disc tissue homogenate liquid prepared in the S3 in the precooled high-speed centrifuge, setting the rotation speed of the centrifuge as 8000-;
s5, taking out the centrifugate in the S4, pouring out the supernatant, and adding a proper amount of trypsin into the supernatant for enzymolysis according to the proportion of 1-5% of the mass of the donkey placenta tissue raw powder to prepare an enzymolysis liquid;
s6, adding a proper amount of TCA-DOC trace protein precipitation concentration reagent into the enzymatic hydrolysate prepared in the S5 according to the proportion that the solid-to-liquid ratio is 1-5% to prepare a donkey placenta tissue protein concentrated solution, placing the tissue concentrated solution into a precooled high-speed centrifuge for centrifugation, setting the rotation speed of the centrifuge as 8000-12000r/min, setting the centrifugation time as 20-30min, and pouring out the supernatant after the centrifugation is finished;
s7, performing fishy smell removing and filtering treatment on the donkey placenta tissue protein concentrated supernatant obtained in the S6 by adopting a fishy smell removing and filtering integrated device, adding 95% ethanol with the volume being 3 times that of the filtered filtrate, performing reduced pressure concentration on the mixed solution by adopting a rotary evaporator, and removing an organic solvent to obtain yellowish transparent liquid;
s8, placing the yellowish transparent liquid prepared in the S7 into a vacuum freeze dryer for freeze drying treatment, and freeze drying for 24-48h to obtain the donkey placentin crystal.
A method for removing fishy smell and filtering donkey placenta tissue protein concentrated solution comprises the following steps:
a1, pouring the supernatant prepared in the S6 into a deodorization device from a feeding hole, and directly flowing the supernatant into a filter box after the adsorption treatment of loose cotton, activated carbon and filter paper;
a2, opening a vacuum suction pump for suction filtration, wherein in the vacuum suction filtration process, the vacuum suction filtration pipeline generates airflow which drives a fan to start rotating, so that a rotating shaft is driven to start rotating, a worm wheel fixedly connected with the rotating shaft starts rotating, a worm meshed with the worm wheel starts rotating, the worm drives a stirring rod to rotate, and filtrate is stirred in the filtrate suction filtration process;
and A3, pouring the filtrate after stirring and suction filtration into a liquid storage tank, opening a liquid discharge valve, and discharging the filtrate from a liquid discharge port.
Further optimized, the freeze-drying time of the fresh donkey placenta tissue in the S1 is not more than 48 h.
Further optimally, the high-speed centrifuge precools the temperature to 4 ℃ in the S4.
Further preferably, the time for enzymolysis by the trypsin added in S5 is 2 h.
And further optimally, the pore diameter of the microporous filter membrane in the S7 is less than or equal to 5 mu m.
Further preferably, the temperature of the decompression concentration of the mixed solution in the S7 is not more than 37 ℃.
The equipment is applied to any method for extracting donkey placenta from a fresh donkey placenta, the deodorization and filtration integrated device comprises a tank body, a filter box and a flip cover, and further comprises a deodorization device, a liquid discharge port is formed in the right side of the lower end of the tank body, a liquid discharge valve is fixedly mounted on the right side of the liquid discharge port, a liquid storage tank is arranged at the lower end inside the tank body, the middle of the upper end of the liquid storage tank is fixedly connected with the bottom of the filter box, the top end of the filter box is fixedly connected with the bottom of the deodorization device, a bearing is arranged between the upper side of the filter box and the lower side of the deodorization device, a feed inlet is formed in the right side of the upper end of the tank body, the lower end of the feed inlet penetrates through the wall surface of the right side of the tank body and is fixedly connected with the right side of the upper end;
loose cotton is arranged at the upper end inside the deodorization device, active carbon is arranged in the middle inside the deodorization device, the lower end in the deodorization device is provided with filter paper, the upper end of the right side of the liquid storage tank is provided with a vacuum pumping and filtering pump, the left side of the vacuum filtration pump is fixedly connected with the right side of the vacuum filtration pipeline, the left side of the vacuum filtration pipeline penetrates through the middle part of the filtration tank and is fixedly connected with the upper end of the left side of the liquid storage tank, the bottom end in the filter box is provided with a microporous filter membrane, the middle in the vacuum filtration pipeline is provided with a fan, the middle part of the fan is fixedly connected with one end of a rotating shaft, the other end of the rotating shaft is fixedly connected with the middle part of a worm wheel, the right side of the worm wheel is meshed and connected with the left side of the worm, the upper end of the worm is movably connected with the axle center of the lower end of the bearing, the lower end of the worm penetrates through the middle of the vacuum filtration pipeline and extends to the lower end inside the filter box, and stirring rods are fixedly mounted on the left side and the right side of the lower end of the worm.
Compared with the prior art, the invention provides a donkey skin treatment method and a process flow before donkey-hide gelatin decoction, and the method has the following beneficial effects:
(1) the method adopts the fresh donkey placenta, and the placentin has the characteristics of high purity, high quality and high safety, so that in the process of extracting the placentin, firstly, the safety inspection needs to be carried out on the raw materials to ensure that the raw materials are not polluted and do not contain any possible pathogenic factors, such as virus, syphilis, HIV and other dangerous factors, therefore, the method carries out quarantine on the live donkey before the raw materials are obtained, the live donkey placenta is directly taken out for tissue in the process of killing the live donkey, the biological activity of the tissue of the fresh donkey placenta is high, the placentin is easy to extract, the donkey placenta is washed clean and then frozen and dried for later use, the method is always carried out under the aseptic condition, and the biological activity of trypsin is favorably kept;
(2) the method of the invention adopts the common freeze-drying method to pretreat the donkey placenta tissue, the main components of the placenta generally comprise protein, nucleic acid, phospholipids, polysaccharose, amino acid, mineral substances, vitamins and the like, the essence of the placenta is micromolecular peptide substances and nucleic acid fragments, and the placenta essence is also the basic element of activated cells, therefore, impurities are required to be removed while extracting the placenta, and the activity of effective components is kept, the placenta raw materials are pretreated by adopting a cooking method in the prior art, the effective components in the placenta are damaged by high temperature, therefore, the method of the invention adopts the common freeze-drying method to pretreat the donkey placenta tissue, and precools a high-speed centrifuge in the extraction process of the donkey placenta, which is beneficial to solid-liquid separation, after the deodorization treatment, a rotary evaporator is adopted to perform reduced pressure concentration on the treatment liquid, the extraction efficiency of donkey placentin can be effectively improved;
(3) the method adopts a deodorization and filtration integrated device to carry out deodorization and filtration treatment on the donkey placenta tissue protein concentrated supernatant, and the traditional placenta extraction deodorization operation is mainly that active carbon is directly added into the donkey placenta tissue protein concentrated supernatant, then the active carbon is filtered out, and then filtrate is filtered by a microporous filter membrane, so that the operation is complicated, the time is wasted, and the used active carbon can not be reused, thereby causing the waste of resources, therefore, the method is provided with the deodorization and filtration integrated device, the prepared donkey placenta tissue protein concentrated supernatant is poured into the deodorization device from a feed inlet, the supernatant directly flows into a filter box through the adsorption treatment of bulky cotton, the active carbon and filter paper, a vacuum suction pump is opened for suction filtration, in the vacuum suction filtration process, a vacuum suction filtration pipeline generates air flow, and the air flow drives a fan to start rotating, the rotating shaft is driven to rotate, the worm wheel fixedly connected with the rotating shaft is driven to rotate, the worm meshed with the worm wheel starts to rotate, the worm drives the stirring rod to rotate, filtrate is stirred in the filtrate filtering process, the filtrate subjected to stirring and filtering flows into the liquid storage tank, the liquid discharge valve is opened, and the filtrate is discharged from the liquid discharge port.
Drawings
FIG. 1 is a schematic flow chart of a process and a method for extracting donkey placentin from fresh donkey placenta according to the present invention;
FIG. 2 is a schematic structural diagram of the deodorization and filtration integrated device of the process and method for extracting donkey placenta from fresh donkey placenta according to the present invention;
fig. 3 is a perspective structure schematic diagram of the deodorization and filtration integrated device of the process and method for extracting donkey placenta from fresh donkey placenta provided by the invention.
The reference numbers in the figures illustrate:
1 tank body, 2 liquid discharging ports, 3 liquid discharging valves, 4 filter boxes, 5 activated carbon, 6 material inlet ports, 7 ultraviolet disinfection lamps, 8 turning covers, 9 fluffy cotton, 10 filter paper, 11 microporous filter membranes, 12 deodorization devices, 13 worm wheels, 14 rotating shafts, 15 fans, 16 worms, 17 stirring rods, 18 vacuum pumping pumps, 19 vacuum pumping pipelines, 20 bearings and 21 liquid storage tanks.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
In the description of the present invention, it is to be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are merely for convenience in describing the present invention and simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the present invention.
Example 1: a process and method for extracting donkey placentin from fresh donkey placenta comprise the following steps:
s1, quarantining the live donkeys and then sending the live donkeys into a large slaughter house for slaughtering, after slaughtering the live donkeys, sending the live donkeys into a food-grade production and processing environment, quickly taking down fresh donkey placenta tissues from the abdomen of a mother donkey and cleaning the fresh donkey placenta tissues in the slaughtering process, and then putting the donkey placenta tissues into a vacuum freeze dryer for freeze preservation for later use;
s2, taking out the donkey placenta tissue freeze-dried in S1, and crushing the donkey placenta tissue by using a tissue crusher under the aseptic condition to prepare donkey placenta tissue raw powder;
s3, under the aseptic condition, weighing a proper amount of donkey placenta tissue raw powder in the S2, adding a proper amount of sterilized deionized water into the donkey placenta tissue raw powder according to the mass ratio of 1:1-1:5, and homogenizing by a homogenizer to prepare donkey placenta tissue homogenate;
s4, opening the high-speed centrifuge for precooling, placing the donkey fetal disc tissue homogenate liquid prepared in the S3 in the precooled high-speed centrifuge, setting the rotation speed of the centrifuge as 8000-;
s5, taking out the centrifugate in the S4, pouring out the supernatant, and adding a proper amount of trypsin into the supernatant for enzymolysis according to the proportion of 1-5% of the mass of the donkey placenta tissue raw powder to prepare an enzymolysis liquid.
The freeze-drying time of the fresh donkey placenta tissues in the S1 is not more than 48 h.
In S4, the temperature is precooled to 4 ℃ by a high-speed centrifuge.
The time for enzymolysis by trypsin added in S5 is 2 h.
The aperture of the microporous filter membrane in S7 is less than or equal to 5 μm.
The concentration temperature of the mixed solution in the S7 is not more than 37 ℃ under reduced pressure.
The method adopts the fresh donkey placenta, and the placentin has the characteristics of high purity, high quality and high safety, so that in the process of extracting the placentin, firstly, the safety inspection needs to be carried out on the raw materials to ensure that the raw materials are not polluted and do not contain any possible pathogenic factors, such as virus, syphilis, AIDS virus and other dangerous factors, therefore, the method carries out quarantine on the live donkey before the raw materials are obtained, the live donkey placenta is directly taken out for tissue in the process of killing the live donkey placenta, the biological activity of the tissue of the fresh donkey placenta is high, the placentin is easy to extract, the donkey placenta is washed clean and then is frozen and dried for later use, the method is always carried out under the aseptic condition, and the biological activity of trypsin is favorably maintained.
Example 2: based on the embodiment 1, but different from the embodiment, according to the proportion that the solid-to-liquid ratio is 1%, adding a proper amount of TCA-DOC micro-protein precipitation concentration reagent into the prepared enzymatic hydrolysate to prepare a donkey placenta tissue protein concentrated solution, placing the tissue concentrated solution into a precooled high-speed centrifuge for centrifugation, setting the rotating speed of the centrifuge to be 8000r/min and the centrifugation time to be 20min, pouring out supernatant after the centrifugation is finished, adopting a deodorization and filtration integrated device to carry out deodorization treatment on the prepared supernatant, carrying out ultrafiltration on the filtrate obtained after the treatment by using a microporous filter membrane, adding 95% ethanol with the volume being 3 times of that of the filtrate obtained after the ultrafiltration, adopting a rotary evaporator to carry out reduced pressure concentration on the mixed solution, removing an organic solvent to obtain yellowish transparent liquid, placing the obtained yellowish transparent liquid into a vacuum freeze dryer for freeze drying, freeze drying for 24 hr to obtain donkey placentin crystal.
The method of the invention adopts the common freeze-drying method to pretreat the donkey placenta tissue, the main components of the placenta generally comprise protein, nucleic acid, phospholipids, polysaccharose, amino acid, mineral substances, vitamins and the like, the essence of the placenta is micromolecular peptide substances and nucleic acid fragments, and the placenta essence is also the basic element of activated cells, therefore, impurities are required to be removed while extracting the placenta, and the activity of effective components is kept, the placenta raw materials are pretreated by adopting a cooking method in the prior art, the effective components in the placenta are damaged by high temperature, therefore, the method of the invention adopts the common freeze-drying method to pretreat the donkey placenta tissue, and precools a high-speed centrifuge in the extraction process of the donkey placenta, which is beneficial to solid-liquid separation, after the deodorization treatment, a rotary evaporator is adopted to perform reduced pressure concentration on the treatment liquid, can effectively improve the extraction efficiency of donkey placentin.
Example 3: a method for removing fishy smell and filtering donkey placenta tissue protein concentrate comprises the following steps:
a1, pouring the supernatant prepared in the S6 into a deodorization device from a feeding hole, and directly flowing the supernatant into a filter box after the adsorption treatment of loose cotton, activated carbon and filter paper;
a2, opening a vacuum suction pump for suction filtration, wherein in the vacuum suction filtration process, the vacuum suction filtration pipeline generates airflow which drives a fan to start rotating, so that a rotating shaft is driven to start rotating, a worm wheel fixedly connected with the rotating shaft starts rotating, a worm meshed with the worm wheel starts rotating, the worm drives a stirring rod to rotate, and filtrate is stirred in the filtrate suction filtration process;
and A3, pouring the filtrate after stirring and suction filtration into a liquid storage tank, opening a liquid discharge valve, and discharging the filtrate from a liquid discharge port.
A device for extracting donkey placenta from a fresh donkey placenta disc comprises a tank body 1, a filter tank 4, a flip cover 8 and a deodorization device 12, wherein a liquid discharging port 2 is formed in the right side of the lower end of the tank body 1, a liquid discharging valve 3 is fixedly mounted on the right side of the liquid discharging port 2, a liquid storage tank 21 is arranged at the lower end inside the tank body 1, the middle of the upper end of the liquid storage tank 21 is fixedly connected with the bottom of the filter tank 4, the top end of the filter tank 4 is fixedly connected with the bottom end of the deodorization device 12, a bearing 20 is arranged between the upper side of the filter tank 4 and the lower side of the deodorization device 12, a feed inlet 6 is formed in the right side of the upper end of the tank body 1, the lower end of the feed inlet 6 penetrates through the right side wall surface of the tank body 1 and is fixedly;
the upper end in the fishy smell removing device 12 is provided with bulky cotton 9, the middle in the fishy smell removing device 12 is provided with activated carbon 5, the lower end in the fishy smell removing device 12 is provided with filter paper 10, the upper end on the right side of the liquid storage tank 3 is provided with a vacuum suction pump 18, the left side of the vacuum suction pump 18 is fixedly connected with the right side of a vacuum suction filtering pipeline 19, the left side of the vacuum suction filtering pipeline 19 penetrates through the middle of the filter tank 4 and is fixedly connected with the upper end on the left side of the liquid storage tank 21, the bottom end in the filter tank 4 is provided with a microporous filter membrane 11, the middle in the vacuum suction filtering pipeline 19 is provided with a fan 15, the middle of the fan 15 is fixedly connected with one end of a rotating shaft 14, the other end of the rotating shaft 14 is fixedly connected with the middle of a worm wheel 13, the right side of the worm wheel 13 is meshed with the left side of a.
The deodorization and filtration integrated device adopted by the method of the invention is used for deodorization and filtration treatment of donkey placenta tissue protein concentrated supernatant, and the traditional placenta extraction and deodorization operation mainly comprises the steps of directly adding active carbon into the donkey placenta tissue protein concentrated supernatant, filtering the active carbon out, and then filtering filtrate by using a microporous filter membrane, so that the operation is complicated, the time is wasted, and the used active carbon can not be recycled, thereby causing the waste of resources.
Example 4: based on the embodiments 1, 2 and 3, but different from the embodiments, live donkeys are quarantined and then sent to a large slaughter house for slaughtering, live donkeys are slaughtered and then enter a food-grade production and processing environment, in the slaughtering process, fresh donkey placenta tissues are quickly taken down from the abdomen of a mother donkey and cleaned, the fresh donkey placenta tissues are crushed by a tissue crusher under the aseptic condition to prepare donkey placenta tissue raw powder, a proper amount of sterilized deionized water is added into the donkey placenta tissue raw powder according to the mass ratio of 1:3, a homogenizer is adopted for homogenizing to prepare donkey placenta tissue homogenate, the donkey placenta tissue homogenate is placed in a precooled high-speed centrifuge after the donkey placenta tissue homogenate is placed in the precooled high-speed centrifuge, the rotation speed of the centrifuge is set to 10000r/min, the centrifugation time is 25min for full centrifugation, supernatant is poured out, a proper amount of trypsin is added into the supernatant according to the mass ratio of 2% of the donkey placenta tissue raw powder for enzymolysis, preparing an enzymatic hydrolysate, adding a proper amount of TCA-DOC trace protein precipitation concentration reagent into the prepared enzymatic hydrolysate according to the proportion that the solid-to-liquid ratio is 2%, preparing a donkey placenta tissue protein concentrated solution, placing the tissue concentrated solution into a precooled high-speed centrifuge for centrifugation, setting the rotation speed of the centrifuge to 10000r/min, setting the centrifugation time to 25min, pouring out a supernatant after the centrifugation is finished, adopting a deodorization and filtration integrated device to prepare the supernatant for deodorization treatment, carrying out ultrafiltration on the treated filtrate by using a microporous membrane, adding 95% ethanol with the volume of 3 times of that of the ultrafiltered filtrate, carrying out reduced pressure concentration on the mixed solution by using a rotary evaporator, removing an organic solvent to obtain a yellowish transparent liquid, placing the yellowish transparent liquid into a vacuum freeze dryer for freeze drying, and carrying out freeze drying for 30h to prepare the donkey placenta crystals.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims (7)

1. A method for extracting donkey placentin from a fresh donkey placenta is characterized by comprising the following steps:
s1, quarantining the live donkeys and then sending the live donkeys into a large slaughter house for slaughtering, after slaughtering the live donkeys, sending the live donkeys into a food-grade production and processing environment, quickly taking down fresh donkey placenta tissues from the abdomen of a mother donkey and cleaning the fresh donkey placenta tissues in the slaughtering process, and then putting the donkey placenta tissues into a vacuum freeze dryer for freeze preservation for later use;
s2, taking out the donkey placenta tissue freeze-dried in S1, and crushing the donkey placenta tissue by using a tissue crusher under the aseptic condition to prepare donkey placenta tissue raw powder;
s3, under the aseptic condition, weighing a proper amount of donkey placenta tissue raw powder in the S2, adding a proper amount of sterilized deionized water into the donkey placenta tissue raw powder according to the mass ratio of 1:1-1:5, and homogenizing by a homogenizer to prepare donkey placenta tissue homogenate;
s4, opening the high-speed centrifuge for precooling, placing the donkey fetal disc tissue homogenate liquid prepared in the S3 in the precooled high-speed centrifuge, setting the rotation speed of the centrifuge as 8000-;
s5, taking out the centrifugate in the S4, pouring out the supernatant, and adding a proper amount of trypsin into the supernatant for enzymolysis according to the proportion of 1-5% of the mass of the donkey placenta tissue raw powder to prepare an enzymolysis liquid;
s6, adding a proper amount of TCA-DOC trace protein precipitation concentration reagent into the enzymatic hydrolysate prepared in the S5 according to the proportion that the solid-to-liquid ratio is 1-5% to prepare a donkey placenta tissue protein concentrated solution, placing the tissue concentrated solution into a precooled high-speed centrifuge for centrifugation, setting the rotation speed of the centrifuge as 8000-12000r/min, setting the centrifugation time as 20-30min, and pouring out the supernatant after the centrifugation is finished;
s7, performing fishy smell removing and filtering treatment on the donkey placenta tissue protein concentrated supernatant obtained in the S6 by adopting a fishy smell removing and filtering integrated device, adding 95% ethanol with the volume being 3 times that of the filtered filtrate, performing reduced pressure concentration on the mixed solution by adopting a rotary evaporator, and removing an organic solvent to obtain yellowish transparent liquid;
the deodorization and filtration integrated device comprises a tank body (1), a filter box (4), a flip cover (8) and a deodorization device (12), wherein a liquid discharge port (2) is formed in the right side of the lower end of the tank body (1), a liquid discharge valve (3) is fixedly mounted on the right side of the liquid discharge port (2), a liquid storage tank (21) is arranged at the lower end inside the tank body (1), the middle part of the upper end of the liquid storage tank (21) is fixedly connected with the bottom of the filter box (4), the top end of the filter box (4) is fixedly connected with the bottom end of the deodorization device (12), a bearing (20) is arranged between the upper side of the filter box (4) and the lower side of the deodorization device (12), a feed inlet (6) is formed in the right side of the upper end of the tank body (1), the lower end of the feed inlet (6) penetrates through the right side wall surface of the tank body (1) and, the bottom of the flip cover (8) is provided with an ultraviolet disinfection lamp (7);
the deodorization device is characterized in that bulky cotton (9) is arranged at the upper end inside the deodorization device (12), activated carbon (5) is arranged in the middle inside the deodorization device (12), filter paper (10) is arranged at the lower end inside the deodorization device (12), a vacuum pumping pump (18) is arranged at the upper end of the right side of the liquid storage tank (21), the left side of the vacuum pumping pump (18) is fixedly connected with the right side of a vacuum pumping pipeline (19), the left side of the vacuum pumping pipeline (19) penetrates through the middle of the filter box (4) and is fixedly connected with the upper end of the left side of the liquid storage tank (21), a microporous filter membrane (11) is arranged at the bottom end inside the filter box (4), a fan (15) is arranged in the middle inside the vacuum pumping pipeline (19), the middle of the fan (15) is fixedly connected with one end of a rotating shaft (14), the other end of the rotating shaft (14) is fixedly connected with the middle of, the upper end of the worm (16) is movably connected with the axis position of the lower end of the bearing (20), the lower end of the worm (16) penetrates through the middle part of the vacuum filtration pipeline (19) and extends to the lower end inside the filter box (4), and stirring rods (17) are fixedly arranged on the left side and the right side of the lower end of the worm (16);
s8, placing the yellowish transparent liquid prepared in the S7 into a vacuum freeze dryer for freeze drying treatment, and freeze drying for 24-48h to obtain the donkey placentin crystal.
2. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which further comprises a method for deodorizing and filtering donkey placenta tissue protein concentrate, and the method comprises the following steps:
a1, pouring the supernatant prepared in the S6 into a deodorization device from a feeding hole, and directly flowing the supernatant into a filter box after the adsorption treatment of loose cotton, activated carbon and filter paper;
a2, opening a vacuum suction pump for suction filtration, wherein in the vacuum suction filtration process, the vacuum suction filtration pipeline generates airflow which drives a fan to start rotating, so that a rotating shaft is driven to start rotating, a worm wheel fixedly connected with the rotating shaft starts rotating, a worm meshed with the worm wheel starts rotating, the worm drives a stirring rod to rotate, and filtrate is stirred in the filtrate suction filtration process;
and A3, pouring the filtrate after stirring and suction filtration into a liquid storage tank, opening a liquid discharge valve, and discharging the filtrate from a liquid discharge port.
3. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which is characterized in that: the freeze-drying time of the fresh donkey placenta tissue in the S1 is not more than 48 h.
4. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which is characterized in that: and pre-cooling the temperature of the high-speed centrifuge in the S4 to 4 ℃.
5. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which is characterized in that: the enzymolysis time of the trypsin added in the S5 is 2 h.
6. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which is characterized in that: the aperture of the microporous filter membrane in the S7 is less than or equal to 5 mu m.
7. The method for extracting donkey placentin from fresh donkey placenta as claimed in claim 1, which is characterized in that: the concentration temperature of the mixed solution in the S7 is not more than 37 ℃ under reduced pressure.
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