CN102488713A - Method for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrated solution - Google Patents

Abstract

The invention discloses a method for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate by using sheep placenta, belonging to the technical field of sheep placenta utilization. The invention uses commercially available freeze-dried sheep placenta as a raw material, first prepares sheep placenta powder, then prepares a crude extract of sheep placenta, and then prepares sheep placenta and sheep placenta hydrolyzed collagen concentrated liquid products. The invention has the advantages of simple method, simple operation, simple equipment, mild reaction conditions, and energy saving; while the sheep placenta is prepared by the method of the invention, various products such as sheep placenta hydrolyzed collagen concentrate and feed additives are also produced, realizing the realization of Comprehensive utilization of sheep placenta; full use of material resources in the production process, no "three wastes" discharge, conducive to environmental protection; low production cost, easy to popularize and apply. The invention can be widely used for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate, and the products prepared by adopting the invention can be widely used in industries such as cosmetics and health care products.

Description

A method for preparing sheep placenta and sheep placenta hydrolyzed collagen concentrate

1. Technical field

The invention belongs to the technical field of utilization of sheep placenta, and in particular relates to a method for preparing sheep placenta element and sheep placenta hydrolyzed collagen concentrate by using sheep placenta.

2. Background technology

Sheep placenta refers to the general term for a variety of active substances extracted from sheep placenta, including a variety of proteins, essential amino acids, nucleic acids, hormones, collagen, immune factors, calcium, iron and zinc and other nutrients. Kidney, regulating immunity, anti-aging, whitening and rejuvenating skin, maintaining body shape and many other health functions. It is clinically used to prevent liver cirrhosis, improve human immunity, treat viral hepatitis, leukopenia, myasthenia gravis and other immune diseases and malignant tumors. It is also a raw material for cosmetics and tonics with magical effects.

Existing methods for preparing sheep placenta, such as the patent "Method for Extracting Sheep Placenta from Sheep Placenta Using Liquid Nitrogen Low-temperature Exhaust Oxygen" with the publication number 101658534A published on March 3, 2010, the disclosed method is: Liquid nitrogen is added to the sheep placenta at the same time, and the ambient temperature is 24 ° C to 25 ° C; the sheep placenta slurry is subjected to enzymatic hydrolysis for 60 minutes at a temperature of 40 ° C, and then extracted with physiological saline, filtered with inorganic membranes, and ultra-high pressure sterilized. The main disadvantages of this method are: 1. The waste residue and waste liquid produced in the production process are not used, which is not conducive to environmental protection; The natural peptide actives in sheep placenta, such as hormones, collagen, and immune factors, are separated and then enzymatically hydrolyzed, and the protease also hydrolyzes the above-mentioned actives, so this method destroys the above-mentioned peptide actives, and the produced products have low biological activity; ④ This method only produces one product of sheep placenta, and the remaining substances are not fully utilized, and the utilization rate of raw materials is not high.

3. Contents of the invention

The purpose of the present invention is to provide a method for preparing sheep placenta and sheep placenta hydrolyzed collagen concentrated solution by using sheep placenta to solve the shortcomings of existing sheep placenta preparation. The method has the characteristics of simple processing technology, common and easy-to-obtain equipment, simple and practical operation steps, high comprehensive utilization rate of resources, environmental protection and no pollution, and the like.

Mechanism of the present invention is: the dried sheep placenta that freezes contains a large amount of nutrients such as water-soluble hormone, collagen, immune factor, essential amino acid, nucleic acid, and calcium, iron and zinc, and can be extracted with aqueous solution; Large molecular weight collagen in the extract, Immune factors have poor stability and are prone to denaturation and precipitation; ultrafiltration is a new technology for separation based on molecular weight differences, which can separate solutes with molecular weight differences exceeding a certain range; when macromolecules in aqueous solution pass through narrow holes under high pressure, they will be sheared, Impact and other forces break into smaller and uniform fragments, which can exist stably in aqueous solution; a large amount of tissue protein remains in the residue of placenta extraction, which can be specifically hydrolyzed into water-soluble collagen by protease; it contains placenta The extract solution of nutrient substance and the aqueous solution containing collagen are vacuum-concentrated to prepare corresponding concentrated solutions respectively.

The purpose of the present invention is achieved in this way: a method for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate: using commercially available freeze-dried sheep placenta as raw material, first preparing sheep placenta powder, and then preparing sheep placenta crude extract, Then prepare sheep placenta extract and sheep placenta hydrolyzed collagen concentrate. Its specific method steps are as follows:

(1) Prepare sheep placenta powder

Using commercially available freeze-dried sheep placenta as raw material, first place the freeze-dried sheep placenta in a pulverizer with a cooling jacket, and then pour cooling water at 4 to 10°C into the jacket of the pulverizer, at a speed of 1000 The pulverization is carried out for 5-15 minutes under the condition of ~5000 rpm. After the pulverization is completed, sieve with a 40-80 mesh sieve, and collect the sifted powder and unsifted particles respectively. The collected sieved powder is used in the next step to prepare the crude sheep placenta extract; the unsifted particles are returned to the pulverizer for further pulverization.

(2) Preparation of Sheep Placenta Crude Extract

After the (1) step was completed, the mass (g) of crossing the 40～80 mesh sieve powder collected according to the step (1): mass concentration was that the volume (mL) ratio of the NaCl solution of 0.05～0.5% was 1: Add the sieved powder into the NaCl solution at a ratio of 50 to 80, stir and mix evenly, and then carry out stirring and extraction at a temperature of 4 to 8°C for 4 to 5 hours. After the extraction is completed, the extract is pumped into a tubular centrifuge, centrifuged at a speed of 6,000 to 10,000 rpm, and the centrifuged liquid and centrifuged sediment are collected respectively. The collected centrifugal supernatant is used to prepare sheep placenta; the collected centrifugal precipitate is used to prepare sheep placenta hydrolyzed collagen concentrate.

(3) Preparation of sheep placenta

After step (2) is completed, pump the centrifuge liquid collected in step (2) into an ultrafiltration machine with a molecular weight cut-off of 2000-10000 Da, and perform ultrafiltration separation at 0.05-0.2 MPa until the volume of the centrifuge liquid is When it is reduced to 1/3-1/6 of the original volume, collect the ultrafiltration filtrate and ultrafiltration retentate respectively. The collected ultrafiltration filtrate can be reused; pump the collected ultrafiltration retentate into a high-pressure homogenizer, perform high-pressure homogenization at 20-60 MPa, collect the pumped liquid, and prepare a high-pressure homogenizer liquid;

After the collected pump out liquid (i.e. high-pressure homogeneous liquid) is combined with the collected ultrafiltration filtrate, it is pumped into the decolorization kettle, according to the quality (g) of activated carbon: the volume (mL) ratio of the combined solution is 1 : the ratio of 10 to 30, adding activated carbon to the combined solution, stirring and decolorizing at room temperature for 10 to 20 minutes, then pumping it into a vacuum filter, performing vacuum filtration, and collecting the filtrate and filter residue respectively. The collected filter residue is sent to the activated carbon regeneration plant for regeneration and can be reused;

Then the collected filtrate is pumped into the reverse osmosis device for reverse osmosis concentration until the soluble solids content in the reverse osmosis retentate reaches 5-8%, and the reverse osmosis permeate and reverse osmosis retained liquid. For the collected reverse osmosis permeate, it can be used for (2) step to extract sheep placenta again; The volume ratio is 1: 0.05-0.08: 0.08-0.1: 0.005-0.01. Add hyaluronic acid, glycerin and antiseptic preservative to the reverse osmosis retentate, stir and mix evenly, and then pump it into the microporous filter. Use a 0.22-0.45um microporous membrane for microfiltration to remove microorganisms and insoluble impurities. After microfiltration is completed, the microfiltration filtrate and microfiltration retentate are collected separately. The collected microfiltration retentate is discharged after being treated up to the standard; the collected microfiltration filtrate is the prepared sheep placenta.

(4) Preparation of sheep placenta hydrolyzed collagen concentrate

After the (3) step is completed, according to the mass (g) of the centrifugal precipitate collected in the (2) step: the volume (mL) ratio of pure water is 1: 3～8 ratio, in the described centrifugal precipitate, add pure water, and pumped into a high-speed homogenizer for beating, and then sieved with a 40-80 mesh sieve to collect the sieved liquid and the unsifted slag (ie, the fascia of sheep placenta). For the unsifted slag, according to the ratio of the mass (g) of the unsifted slag: the volume (mL) of pure water detergent is 1:30, the unsifted slag is washed with pure water, respectively Collect the washing liquid and washing residue. The washing slag collected at the end is discharged after being treated up to the standard; finally, the sieved liquid and the washing liquid are combined to prepare a placental residue homogenate.

Then pump the placenta residue homogenate into the reaction kettle, stir and heat up to 50-60°C, and adjust its pH value to 8-9 with sodium hydroxide solution, then according to the volume of the placenta residue homogenate pumped into the reaction kettle (mL ): the mass (g) of alkaline protease: the mass (g) of neutral protease: the mass (g) ratio of acidic protease is the ratio of 1: 0.02～0.03: 0.02～0.03: 0.02～0.03, successively in the reactor Add alkaline protease, neutral protease and acid protease respectively, and stir and hydrolyze at a constant temperature of 50-60° C. for 40-60 minutes respectively. After the continuous hydrolysis is completed, the hydrolyzate is pumped into the tubular centrifuge, and centrifuged at a speed of 3000-8000 rpm, and the centrifuge supernatant and centrifuge sediment are collected respectively. For the collected centrifuge, it can be used as feed additive after being dried and pulverized; for the collected centrifuge, it is pumped into the decolorization kettle, according to the mass (g) of activated carbon: the volume (mL) ratio of the combined solution is 1: Add activated carbon at a ratio of 10 to 30, and heat up to 80 to 100°C, stir and decolorize for 10 to 20 minutes, then pump the decolorized solution into a suction filter, perform vacuum filtration, and collect the filtrate and filter residue separately. The collected filter residue is sent to the activated carbon regeneration plant for regeneration and can be reused.

Finally, pump the collected filtrate into a vacuum concentrator, and carry out vacuum concentration at a vacuum degree of 0.04-0.08MPa and a temperature of 60-100°C until the concentration of soluble solids in the concentrate reaches 20%-25% %, collect the vacuum concentrate. For the collected vacuum concentrate, pump it into the preparation kettle, according to the volume ratio of vacuum concentrate: glycerin: 100 mold antiseptics: sodium carboxymethyl cellulose is 1: 0.08～0.1: 0.005～0.01: 0.01～0.03 According to the ratio, glycerin, chlorothalonide preservative and sodium carboxymethyl cellulose are added into a mixing kettle, stirred and mixed evenly, and the sheep placenta hydrolyzed collagen concentrated solution with a short peptide content of 15% to 20% is prepared.

After the present invention adopts above-mentioned technical scheme, mainly have following effect:

1. The method of the present invention uses simple techniques such as NaCl solution extraction, centrifugation, enzymatic hydrolysis, microporous filtration, ultrafiltration and vacuum concentration in the production process, easy to operate, simple equipment, mild reaction conditions, energy saving, low production cost, and convenient Promote apps.

2. The method of the present invention realizes the comprehensive utilization of sheep placenta in the production process. When the sheep placenta extract is prepared in the invention, by-products such as sheep placenta hydrolyzed collagen concentrate and feed additives are also obtained, the obtained products are many, the yield is high, and the production cost is further reduced.

3. The materials in the process of the method of the present invention are all recycled, without the discharge of "three wastes", and the by-products are recycled or discharged after being treated up to the standard, which not only further reduces the production cost, but also protects the environment.

4. The sheep placenta produced by the production process of the present invention retains active substances such as hormones, collagen, and immune factors, and can be used to prepare skin products such as facial masks and eye patches. Function; the produced sheep placenta hydrolyzed collagen concentrate can be added to health care products to enhance resistance and immunity.

The invention can be widely used for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate. The product prepared by the method of the invention can be widely used in industries such as cosmetics and health care products.

4. Specific implementation

The present invention will be further described below in combination with specific embodiments.

Example 1

A method for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate by using sheep placenta, the specific steps are as follows.

(1) Prepare sheep placenta powder

Using commercially available freeze-dried sheep placenta as raw material, first place the freeze-dried sheep placenta in a pulverizer with a cooling jacket, and then pour cooling water at 4°C into the jacket of the pulverizer, at a speed of 1000 rpm The pulverization was carried out for 5 minutes under the condition of minute. After the pulverization is completed, sieve with a 40-mesh sieve, and collect the sieved powder and unsifted particles respectively. The collected sieved powder is used in the next step to prepare the crude sheep placenta extract; the unsifted particles are returned to the pulverizer for further pulverization.

(2) Preparation of Sheep Placenta Crude Extract

After the (1) step was completed, the mass (g) of crossing the 40 mesh sieve powder collected according to the (1) step: the volume (mL) ratio of the NaCl solution with a mass concentration of 0.05% was a ratio of 1:50, The sieved powder was added into the NaCl solution, stirred and mixed evenly, and then stirred and extracted at a temperature of 4° C. for 4 hours. After the extraction is completed, the extract is pumped into a tubular centrifuge, and centrifuged at a speed of 6000 rpm to collect the centrifuge supernatant and centrifuge sediment respectively. The collected centrifugal supernatant is used to prepare sheep placenta; the collected centrifugal precipitate is used to prepare sheep placenta hydrolyzed collagen concentrate.

(3) Preparation of sheep placenta

After step (2) is completed, pump the centrifuge liquid collected in step (2) into an ultrafiltration machine with a molecular weight cut-off of 2000Da, and perform ultrafiltration separation at 0.05MPa until the volume of the centrifuge liquid is reduced to the original volume Up to 1/3 of the time, the ultrafiltration filtrate and the ultrafiltration retentate were collected respectively. The collected ultrafiltration filtrate can be reused; pump the collected ultrafiltration retentate into a high-pressure homogenizer, perform high-pressure homogenization at 20MPa, collect the pumped liquid, and prepare a high-pressure homogeneous liquid;

After the collected pump out liquid (i.e. high-pressure homogeneous liquid) is combined with the collected ultrafiltration filtrate, it is pumped into the decolorization kettle, according to the quality (g) of activated carbon: the volume (mL) ratio of the combined solution is 1 : ratio of 10, add activated carbon to the combined solution, and stir and decolorize at room temperature for 10 minutes. Then pump the decolorized liquid into a vacuum suction filter, carry out vacuum suction filtration, and collect the filtrate and filter residue respectively. The collected filter residue is sent to the activated carbon regeneration plant for regeneration and can be reused;

Then the collected filtrate is pumped into the reverse osmosis device for reverse osmosis concentration until the soluble solid content in the reverse osmosis retentate reaches 5%, and the reverse osmosis permeate and the reverse osmosis retentate are collected respectively. For the collected reverse osmosis permeate, it can be used for (2) step to extract sheep placenta again; The volume ratio is 1: 0.05: 0.08: 0.005. Add hyaluronic acid, glycerin and antiseptic preservative to the reverse osmosis retentate, stir and mix evenly, pump into the microporous filter, and use 0.22um microporous The filter membrane is used for microfiltration to remove microorganisms and insoluble impurities. After microfiltration is completed, the microfiltration filtrate and microfiltration retentate are collected separately. The collected microfiltration retentate is discharged after being treated up to the standard; the collected microfiltration filtrate is the prepared sheep placenta.

(4) Preparation of sheep placenta hydrolyzed collagen concentrate

After the 3rd step is finished, according to the mass (g) of the centrifuge sediment collected in the step (2): the volume (mL) ratio of pure water is the ratio of 1: 3, add pure water in the centrifuge sediment, and pump into After beating in a high-speed homogenizer, sieve with a 40-mesh sampling sieve, and collect the sieved liquid and unsifted slag (ie, the fascia of sheep placenta) respectively. For the unsifted slag, according to the ratio of the mass (g) of the unsifted slag: the volume (mL) of pure water detergent is 1:30, the unsifted slag is washed with pure water, respectively Collect the washing liquid and washing residue. The washing slag collected at the end is discharged after being treated up to the standard; finally, the sieved liquid and the washing liquid are combined to prepare a placental residue homogenate.

Then placenta dregs homogenate is pumped into the reactor, stirred and heated to 50°C, and its pH value is adjusted to 8 with sodium hydroxide solution, then according to the volume (mL) of the placenta dregs homogenate pumped into the reactor: alkaline The quality (g) of protease: the quality (g) of neutral protease: the mass (g) ratio of acidic protease is the ratio of 1: 0.02: 0.02: 0.02, adds alkaline protease, neutral protease, acid protease, and hydrolyzed at a constant temperature of 50°C for 40 minutes. After the continuous hydrolysis is completed, the hydrolyzate is pumped into a tubular centrifuge, and centrifuged at a speed of 3000 rpm to collect the centrifuge supernatant and centrifuge sediment respectively. For the collected centrifuge, it can be used as feed additive after being dried and pulverized; for the collected centrifuge, it is pumped into the decolorization kettle, according to the mass (g) of activated carbon: the volume (mL) ratio of the combined solution is 1: Add activated carbon at a ratio of 10, and raise the temperature to 80°C, stir and decolorize for 10 minutes, then pump the decolorized solution into a suction filter, perform vacuum filtration, and collect the filtrate and filter residue respectively. The collected filter residue is sent to the activated carbon regeneration plant for regeneration and can be reused.

Finally, the collected filtrate is pumped into a vacuum concentrator, and vacuum concentrated at a vacuum of 0.04MPa and a temperature of 60°C until the concentration of soluble solids in the concentrated solution reaches 20%. Concentrate. For the collected vacuum concentrate, it is pumped into the blending kettle, and according to the volume ratio of vacuum concentrate: glycerol: 100 mold preservatives: sodium carboxymethyl cellulose is 1: 0.08: 0.005: 0.01, in the blending kettle Adding glycerin, chlorothalonid preservative and sodium carboxymethyl cellulose, stirring and mixing evenly, the sheep placenta hydrolyzed collagen concentrated solution with a short peptide content of 15% is prepared.

Example 2

A method for preparing sheep placenta extract and sheep placenta hydrolyzed collagen concentrate with sheep placenta, same as embodiment 1, wherein:

In step (1), the cooling water temperature is 8° C., and the pulverization time is 10 minutes. After the pulverization is completed, pass through a 60-mesh sieve.

In the (2) step, the mass (g) of the sieved powder collected in the (1) step: the volume (mL) ratio of 0.2% NaCl solution is 1: 60, the extraction temperature is 6° C., and the extraction time is 4.5 hours. The centrifuge speed is 8000 rpm.

In the (3) step, the molecular weight cut-off of the ultrafiltration membrane is 5000Da, and the ultrafiltration pressure is 0.1MPa. When the ultrafiltration is reduced to 1/5 of the original volume, the high-pressure homogeneous pressure is 40MPa.

The mass (g) of activated carbon: the volume (mL) ratio of the combined liquid is 1:20, the temperature is raised to 90°C, and the mixture is stirred for 15 minutes to decolorize.

The soluble solids content in the reverse osmosis retentate is 7%, the volume ratio of reverse osmosis retentate: 1% hyaluronic acid: glycerin: balmicide preservative is 1: 0.06: 0.09: 0.008, and the microporous membrane aperture is 0.45 um.

In the (4) step, the mass (g) of the centrifuged sediment: the volume (mL) ratio of pure water is 1: 5, and is sieved with a 60-mesh sieve.

Stirring is warming up to 55 DEG C, pH is 8.5, the volume (mL) of the homogenate liquid of the placenta residue that pumps into reactor: the quality (g) of alkaline protease: the quality (g) of neutral protease: the quality (g) of acid protease The ratio of 1:0.025:0.025:0.025 was stirred and hydrolyzed at a constant temperature of 55° C. for 50 minutes, and the speed of the centrifuge was 5000 rpm. The mass (g) of activated carbon: the volume (mL) ratio of the centrifugal supernatant was 1:20, the temperature was raised to 90° C., and the color was decolorized for 15 minutes.

Carrying out the vacuum tightness of vacuum concentration is 0.06MPa, and temperature is 80 ℃, till when the soluble solids concentration reaches 22% in the concentrated solution; The ratio is 1:0.09:0.008:0.02, and the short peptide content in the prepared sheep placenta hydrolyzed collagen concentrate is 17%.

Example 3

A method for preparing sheep placenta and sheep placenta peptide with sheep placenta, same as embodiment 1, wherein:

In the (1) step, the temperature of the cooling water is 10° C., and the pulverization time is 15 minutes. After the pulverization is completed, pass through an 80-mesh sieve.

In the 2nd step, the mass (g) of the powder of sieving: 0.5%NaCl solution volume (mL) ratio is 1: 80, extraction temperature 8 ℃, extraction time 5 hours, centrifuge speed is 10000 rpm.

In the (3) step, the molecular weight cut-off of the ultrafiltration membrane is 10000Da, and the ultrafiltration pressure is 0.2MPa. When the ultrafiltration is reduced to 1/6 of the original volume, the high-pressure homogeneous pressure is 60MPa.

The mass (g) of activated carbon: the volume (mL) ratio of the combined solution is 1:30, the temperature is raised to 100°C, and the mixture is stirred for 20 minutes to decolorize.

The soluble solids content in the reverse osmosis retentate is 8%, the volume ratio of reverse osmosis retentate: 1% hyaluronic acid: glycerin: balmicide preservative is 1: 0.08: 0.1: 0.01, and the pore size of the microporous membrane is 0.45 um.

In the (4) step, the mass (g) of the centrifuged sediment: the volume (mL) ratio of pure water is 1: 8, and is sieved with 80 mesh sieves.

Stirring is heated up to 60 DEG C, pH is 9, the volume (mL) of the placental residue homogenate pumped into reaction kettle: the quality (g) of alkaline protease: the quality (g) of neutral protease: the quality (g) of acid protease The ratio of 1:0.03:0.03:0.03 was stirred and hydrolyzed at a constant temperature of 60° C. for 60 minutes. The rotating speed of the centrifuge is 8000 rpm, the mass (g) of activated carbon: the volume (mL) ratio of the supernatant liquid is 1:30, the temperature is raised to 100° C., and the decolorization takes 20 minutes.

Carrying out the vacuum degree of vacuum concentration is 0.08MPa, and temperature is 100 ℃, till when the concentration of soluble solids in the concentrated solution reaches 25%; The ratio is 1:0.1:0.01:0.03, and the short peptide content in the prepared sheep placenta hydrolyzed collagen concentrate is 20%.

Claims (1)

1. one kind prepares the method for the plain and Placenta caprae seu ovis hydrolytic collagen concentrated solution of Placenta caprae seu ovis with Placenta caprae seu ovis, it is characterized in that the concrete steps of said method are following:

(1) preparation Placenta caprae seu ovis powder

With commercially available lyophilizing Placenta caprae seu ovis is raw material; Earlier the lyophilizing Placenta caprae seu ovis is placed the pulverizer of band cooling jacket; In the chuck of pulverizer, feeding 4～10 ℃ cooling water again, is to pulverize 5～15 minutes under 1000～5000 rev/mins the condition at rotating speed, powder is pure accomplish after; Sieve with 40～80 purpose sub-sieves, collect powder and the unsifted granule that sieves respectively;

(2) the plain crude extract of preparation Placenta caprae seu ovis

After (1) step accomplished, the quality of mistake 40～80 order sub-sieve powder of collecting according to (1) step: mass concentration was that the volume ratio of 0.05～0.5% NaCl solution is the ratio of 1g: 50～80mL, and the powder that will sieve joins in the NaCl solution; After mixing; Under 4～8 ℃ temperature, stir and extracted 4～5 hours again, after extraction is accomplished; Extracting solution is pumped in the tube centrifuge; Under 6000～10000 rev/mins rotating speed, carry out centrifugalize, collect centrifugal clear liquid and centrifugation respectively;

(3) the preparation Placenta caprae seu ovis is plain

(2) step went on foot the centrifugal clear liquid of collecting with (2) and pumps in the ultrafilter that molecular cut off is 2000～10000Da, under 0.05～0.2MPa after accomplishing; Carry out ultra-filtration and separation, till the centrifugal clear liquid volume is reduced to 1/3 of original volume～1/6 o'clock, collect ultrafiltration filtered solution and ultrafiltration trapped fluid respectively; With the ultrafiltration trapped fluid of collecting; Pump in the high pressure homogenizer, under 20～60MPa, carry out high pressure homogenize, collecting pump fluid;

That will collect pumps liquid again, i.e. high pressure homogenize liquid is after the ultrafiltration filtered solution of collecting with (3) step merges; Pump in the decolouring still, according to the quality of active carbon: the volume ratio of amalgamation liquid is the ratio of 1g: 10～30mL, in amalgamation liquid, adds active carbon; At room temperature stir decolouring 10～20 minutes; Then destaining solution is pumped in the vacuum filtration machine, carry out vacuum filtration, collect filtered solution and filtering residue respectively;

Then the filtered solution of collecting is pumped in the counter-osmosis device; Carry out reverse osmosis concentration, till soluble solid content reached to the reverse osmosis trapped fluid, collect reverse osmosis infiltrate liquid and reverse osmosis trapped fluid respectively at 5～8% o'clock; For the reverse osmosis trapped fluid of collecting; According to the reverse osmosis trapped fluid: mass concentration is 1% hyaluronic acid: glycerol: the volume ratio of BAIMEISHA antiseptic is 1: 0.05～0.08: 0.08～0.1: 0.005～0.01 ratio, in the reverse osmosis trapped fluid, adds hyaluronic acid, glycerol and BAIMEISHA antiseptic, after mixing; Pump in the micropore filter; Microporous filter membrane with 0.22～0.45um carries out microfiltration, and microfiltration is collected microfiltration filtered solution and microfiltration trapped substance respectively after accomplishing;

(4) preparation Placenta caprae seu ovis hydrolytic collagen concentrated solution

After (3) step accomplished, the quality of the centrifugation of collecting according to (2) step: the volume ratio of pure water was the ratio of 1g: 3～8mL, in centrifugation, adds pure water; And pump into pull an oar in the high-speed homogenization machine after, sieve with 40～80 purpose sub-sieves, collect sieve liquid and unsifted slag respectively; For unsifted slag; Quality according to unsifted slag: the volume ratio of pure water is the ratio of 1g: 30mL, with pure water unsifted slag is washed, and collects cleaning mixture and washed-residue respectively; To sieve at last liquid and cleaning mixture merges, and promptly prepares Placenta Hominis slag homogenate;

Then Placenta Hominis slag homogenate is pumped in the agitated reactor, stir and be warming up to 50～60 ℃, and its pH value of use sodium hydroxide solution adjusting is 8～9; Again according to the volume that pumps into Placenta Hominis slag homogenate in the agitated reactor: the quality of alkaline protease: the quality of neutral protease: the mass ratio of acid protease is the ratio of 1mL: 0.02～0.03g: 0.02～0.03g: 0.02～0.03g, in agitated reactor, adds alkaline protease, neutral protease, acid protease successively respectively, and under 50～60 ℃ constant temperature, stirs hydrolysis 40～60 minutes respectively; Continuous hydrolysis pumps into hydrolyzed solution in the tube centrifuge, under 3000～8000 rev/mins rotating speed after accomplishing; Carry out centrifugalize, collect centrifugal clear liquid and lower sediment respectively, for the centrifugal clear liquid of collecting; Pump in the decolouring still; Quality according to active carbon: the volume ratio of amalgamation liquid is the ratio of 1mL: 10～30g, adds active carbon, and is warming up to 80～100 ℃; Stir decolouring after 10～20 minutes; Destaining solution is pumped in the sucking filtration machine, carry out vacuum filtration, collect filtered solution and filtering residue respectively;

At last the filtered solution of collecting is pumped in the vacuum decker; In vacuum is 0.04～0.08MPa, and temperature is under 60～100 ℃ the condition, to carry out vacuum concentration; Till soluble solid concentration reached 20%～25% o'clock in concentrated solution; Collect vacuum concentration liquid, the vacuum concentration liquid for collecting pumps in the compounding kettle; According to vacuum concentration liquid: glycerol: BAIMEISHA antiseptic: the volume ratio of sodium carboxymethyl cellulose is 1: 0.08～0.1: 0.005～0.01: 0.01～0.03 ratio; In compounding kettle, add glycerol, BAIMEISHA antiseptic, sodium carboxymethyl cellulose, mix, just prepare small peptide content and be 15%～20% Placenta caprae seu ovis hydrolytic collagen concentrated solution.