CN111925408B - Preparation method of periplaneta americana polypeptide extract taking water as medium - Google Patents
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- CN111925408B CN111925408B CN202010819339.XA CN202010819339A CN111925408B CN 111925408 B CN111925408 B CN 111925408B CN 202010819339 A CN202010819339 A CN 202010819339A CN 111925408 B CN111925408 B CN 111925408B
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Abstract
The invention belongs to the technical field of extraction and preparation of natural components, and relates to a preparation method of a periplaneta americana polypeptide extract, in particular to a preparation method of a periplaneta americana polypeptide extract taking water as a medium, which comprises the following steps of: mixing the periplaneta americana medicinal material with activated carbon powder, and crushing to obtain a crushed material; dispersing the crushed material in water to obtain a dispersion, and grinding the dispersion to obtain an extraction mixture; and purifying and separating the periplaneta americana polypeptide extract from the extraction mixture. The method takes water as a medium, avoids organic solvent residues, complete extraction of polypeptide and low extraction cost, adopts activated carbon to adsorb grease components, avoids the difficult separation of the emulsion of the extracting solution, ensures the smoothness of the process, removes the fishy smell of the product through the adsorption of the activated carbon, improves the quality of the product, and has good market application prospect and potential huge economic benefit.
Description
Technical Field
The invention belongs to the technical field of extraction and preparation of natural components, relates to a preparation method of a periplaneta americana polypeptide extract, and in particular relates to a preparation method of a periplaneta americana polypeptide extract taking water as a medium.
Background
Modern researches show that the American cockroach, also called as Blatta seu Bulgaria, has the main active ingredients of polypeptide and biological enzyme, and has obvious effects of promoting tissue repair and regeneration, resisting tumor and resisting hepatitis. The marketed medicines prepared from the periplaneta americana as the main raw material comprise new rehabilitation liquid, xinmailong, ganlong capsules and the like, and have good economic and social benefits.
The periplaneta americana is an animal medicinal material, is rich in grease, is generally prepared by degreasing an organic solvent (petroleum ether, normal hexane and the like) and then using a high-polarity solvent (alcohol or alcohol-water mixture) as an extraction solvent, and has the defects that: the cost of using the organic solvent is high; the residue of the oil component is large; the fishy smell of the extract is large and is not easy to remove; partial polypeptides have low solubility in organic solvents resulting in incomplete extraction. If water is used for replacing an organic solvent as an extraction solvent, although the solubility of the polypeptide in the extraction solvent can be improved, the solubility of the grease in the water is extremely low, the content of the grease reaches a certain critical value, the phenomenon of emulsification of an extracting solution in the extraction process is extremely easy to occur, and target components (protein, polypeptide, amino acid and the like) cannot be effectively separated from a large amount of grease in medicinal materials, so that the product quality cannot be effectively ensured.
Disclosure of Invention
The invention aims to provide a preparation method of a periplaneta americana polypeptide extract taking water as a medium, which is used for solving the technical problem of emulsification phenomenon when water is used as an extraction solvent.
In order to solve the technical problems, the technical scheme of the invention is as follows:
the preparation method of the periplaneta americana polypeptide extract taking water as a medium comprises the following steps in sequence:
s1: mixing the periplaneta americana medicinal material with activated carbon powder, and crushing to obtain a crushed material;
s2: dispersing the crushed material in water to obtain a dispersion liquid, and homogenizing the dispersion liquid to obtain an extraction mixture;
s3: and purifying and separating the periplaneta americana polypeptide extract from the extraction mixture.
By adopting the technical scheme, the technical principle and the beneficial effects are as follows:
in the process of crushing the medicinal materials, active carbon powder (200-500 meshes) is used for wrapping the powder of the animal medicinal materials, the oil on the micro-powder surface of the animal medicinal materials is isolated and adsorbed, and the crushed materials are obtained after crushing treatment. The crushed materials comprise active carbon powder and crushed periplaneta americana medicinal materials. After the crushing is finished, extracting crushed materials (including active carbon powder) by using water as a medium, dispersing the crushed materials in water to obtain dispersion liquid, and continuously grinding and homogenizing the periplaneta americana medicinal material in the extraction process, so that the medicinal material raw material powder continuously releases oil components and polypeptide components, the oil components are adsorbed by the active carbon (the active carbon also has the effect of assisting in grinding medicinal materials), and the polypeptide components enter the water. And (3) obtaining an extraction mixture after grinding and extracting, wherein the extraction mixture is a dispersion liquid subjected to grinding treatment. Separating residue (containing active carbon powder) and liquid phase from the extract mixture after extraction, and purifying to obtain periplaneta americana polypeptide extract. In the scheme, homogenization refers to homogenizing materials, and the materials are subjected to splitting, crushing, blending and other processes in a container by rotating a rotary knife driven by a motor so as to be stirred and smashed. Besides the rotary knife driven by the motor, the dispersion liquid can be ground by a grinder in the prior art, so that the materials are stirred and smashed, and the homogenization of the materials can be realized. The key point of the scheme is that active carbon powder and animal medicine powder are required to exist simultaneously in the homogenization process, and the active carbon powder plays a role in preventing system emulsification.
In the processes of medicinal material crushing and polypeptide extraction, the addition of the activated carbon powder can fully absorb grease in medicinal materials, so that the phenomenon of emulsification of an extracting solution is prevented, the efficiency of polypeptide extraction is improved, and the product quality is ensured. The periplaneta americana polypeptide extract prepared by the method has the advantages of high yield, high content of functional components and the like. The active carbon is usually used for removing impurities (lipids, pigments and the like) in the extracting solution (after extraction is completed), but in the scheme, the active carbon powder is added into the process in the processes of crushing and extracting the medicinal materials, so that unexpected technical effects are brought, namely the aims of preventing the emulsifying phenomenon of an extracting system and improving the quality of products are achieved.
The invention provides a preparation method for extracting periplaneta americana polypeptide by using water as an extraction medium, which uses water as the medium, avoids organic solvent residues, has complete polypeptide extraction and low extraction cost, uses active carbon to adsorb grease components, avoids the difficult separation of emulsion of an extracting solution, ensures the smoothness of the process, removes the fishy smell of the product by the adsorption of the active carbon, improves the quality of the product, and has good market application prospect and potential huge economic benefit.
Further, in S1, the temperature of the mixture of the periplaneta americana medicinal material raw material and the activated carbon powder is-20-0 ℃.
By adopting the technical scheme, the quality of the protein or the polypeptide can be ensured at low temperature, and the degradation of the protein or the polypeptide is prevented; the occurrence of the rotting phenomenon of the periplaneta americana can be avoided at low temperature.
Further, in S1, the mass ratio of the periplaneta americana medicinal material raw material to the activated carbon powder is 1:1-1:2.
By adopting the technical scheme, the crushed material contains the activated carbon powder as the grease adsorbent, so that the adhesion of the material powder is prevented. The active carbon powder fully acts with oil substances in the process of crushing and extracting medicinal materials, so as to prevent the occurrence of emulsification. Optimization research shows that the ratio of the medicinal materials to the active carbon is lower than 1:1, the mixed powder is easy to adhere to and block the screen (the active carbon powder has the function of preventing the screen from being blocked after the medicinal materials are crushed), and a slight emulsification phenomenon still occurs in the extraction process; and if the ratio is higher than 1:2, the effects of crushing the medicines and adsorbing grease are not improved remarkably, and the waste of the activated carbon materials is caused.
Further, in S1, the periplaneta americana medicinal material and the activated carbon powder are mixed, crushed, and then sieved to obtain a sieved material; the mesh number of the sieve is greater than or equal to 80 mesh.
By adopting the technical scheme, the periplaneta americana medicinal material raw material is crushed to be smaller than a certain particle size, so that medicinal material particles are fully contacted with medium water, and the dissolution efficiency of water to target components is improved.
Further, in S2, the crushed material is dispersed in water to obtain a dispersion liquid, and the dispersion liquid is subjected to a homogenization treatment until the crushed material in the dispersion liquid reaches 200 to 500 meshes, to obtain an extraction mixture.
By adopting the technical scheme, the materials are homogenized until the crushed materials in the dispersion liquid reach 200-500 meshes, so that the functional components can be fully dissolved out, and the purpose components can be fully extracted.
Further, in S2, the homogenization treatment is performed in an environment of 0 to 10 ℃.
By adopting the technical scheme, the animal medicinal material particles have certain granularity hardness in the temperature range, do not soften and agglomerate, and ensure the homogenization efficiency; meanwhile, the liquid medicine is not frozen, so that excessive abrasion of equipment caused by the occurrence of ice particles is avoided.
Further, in S2, the water is used in an amount of: 5-10L of water is used as 1kg of periplaneta americana medicinal material raw material.
By adopting the technical scheme, the sufficient water can fully dissolve out the functional components (substances such as polypeptide and the like) and improve the yield of the target components.
Further, in S3, the extract mixture is first subjected to solid-liquid separation and a liquid phase is taken to obtain a crude extract; then adding active carbon into the crude product of the extracting solution, stirring, and filtering to obtain a liquid phase to obtain a periplaneta americana polypeptide extracting solution; concentrating and drying the periplaneta americana polypeptide extract to obtain the periplaneta americana polypeptide extract.
By adopting the technical scheme, the activated carbon can further remove impurities such as grease, pigment and the like. The activated carbon herein may be a conventional activated carbon for adsorbing impurities, and the aforementioned activated carbon powder is not necessarily used.
Further, in S3, the extract mixture is subjected to solid-liquid separation by centrifugation at 8000 to 25000rpm.
By adopting the technical scheme, the centrifugal rotating speed can lead the solid and the liquid to be fully separated, and particularly the active carbon powder and the target component to be separated.
Further, in S3, the amount of the activated carbon added into the crude extract is 4-6% of the mass of the raw material of the periplaneta americana; adding active carbon into the crude product of the extract, stirring for 30min, and filtering to obtain liquid phase to obtain the periplaneta americana polypeptide extract.
By adopting the technical scheme, the dosage of the active carbon and the stirring time ensure the sufficient removal of impurities such as grease, pigment and the like.
Detailed Description
Example 1
Taking 5kg of periplaneta americana medicinal material, adding 5kg of activated carbon (200-300 meshes, specifically 300 meshes in the embodiment), freezing to-20 ℃, and crushing by a crusher (the temperature of the material is kept below 0 ℃ in the crushing process) to obtain crushed materials (the crushed materials are obtained by sieving with a 80-mesh sieve, and the undersize is obtained). 25L of pure water was added to the pulverized material, and the pulverized material was dispersed in pure water to obtain a dispersion. Homogenizing and extracting (maintaining the temperature of the material at 0-10deg.C during homogenizing) with a homogenizer at 10000rpm until the powder obtained after homogenizing the crushed material reaches 200 mesh to obtain an extraction mixture (i.e. homogenized dispersion). Centrifuging and filtering twice, wherein the first centrifuging speed is set to 8000rpm, the second centrifuging speed is set to 16000rpm, centrifuging and filtering, and collecting liquid phase to obtain crude extract. Degreasing and deodorizing the crude product of the extracting solution twice by using active carbon (200 meshes) (5% of active carbon of the mass of the raw material of the periplaneta americana is added each time and stirred for 30 min), and filtering to obtain a liquid phase to obtain the periplaneta americana polypeptide extracting solution. Concentrating the periplaneta americana polypeptide extract at 70 ℃ under reduced pressure to a small volume, and then drying at 70 ℃ under reduced pressure in vacuum to obtain 952g of periplaneta americana polypeptide extract.
Example 2
Taking 20kg of periplaneta americana medicinal material, adding 30kg of active carbon (400-500 meshes, specifically 500 meshes in the embodiment), freezing to-10 ℃, and crushing into powder by a crusher (the temperature is kept at-5 ℃ in the crushing process) to obtain a crushed material (sieving with a 80-mesh sieve, and taking a screen-undersize product). 200L of water is added into the crushed materials, the mixture is mixed evenly to obtain dispersion liquid, a homogenizer is used for homogenization and extraction (the temperature is kept between 0 and 10 ℃ in the homogenization process) at 10000rpm until the powder reaches 400 meshes, and an extraction mixture (namely the dispersion liquid after grinding treatment) is obtained. The mixture was centrifuged and filtered (rotation speed was set at 25000 rpm) to obtain a liquid phase and a crude extract was obtained. Degreasing and deodorizing the crude product of the extracting solution twice by using active carbon (each time adding active carbon accounting for 6% of the mass of the periplaneta americana medicinal material for removing, stirring for 30 min), filtering, concentrating the filtrate under reduced pressure at 70 ℃ to a small volume, and then freeze-drying to obtain 3660g of periplaneta americana polypeptide extract.
Example 3
Taking 2kg of periplaneta americana medicinal material, adding 4kg of active carbon (400-500 meshes, specifically 500 meshes in the embodiment), freezing to-20 ℃, grinding into powder by a grinder (keeping the temperature below-10 ℃ in the grinding process), and obtaining a crushed material (sieving by a 200-mesh sieve and taking the undersize). Adding 20L of water into the crushed material, uniformly mixing to obtain a dispersion liquid, homogenizing and extracting (maintaining 0-10 ℃ in the homogenizing process) by using a homogenizer at 10000rpm until the powder reaches 500 meshes, and obtaining an extraction mixture (namely the dispersion liquid subjected to grinding treatment). The extract mixture was centrifuged and filtered twice, the first rotation speed was set at 16000rpm and the second rotation speed was set at 25000rpm, and the liquid phase was taken to obtain a crude extract. Degreasing and deodorizing the crude product of the extracting solution twice by using active carbon (4% of active carbon of the mass of the periplaneta americana medicinal material is added each time, stirring is carried out for 30 min), filtering, concentrating the filtrate at 70 ℃ under reduced pressure to a small volume, and then freeze-drying to obtain 406g of periplaneta americana polypeptide extract.
Example 4
Taking 100kg of periplaneta americana medicinal material, adding 100kg of active carbon (200-300 meshes, specifically 300 meshes in the embodiment), freezing to-10 ℃, and crushing into medicinal powder (the medicinal powder is kept below 0 ℃ in the crushing process) to obtain a crushed material (the crushed material is obtained by sieving with a 80-mesh sieve and taking a screen lower). Adding 800L of water into the crushed materials, uniformly mixing to obtain a dispersion liquid, carrying out homogenizing extraction (the temperature is kept between 0 and 10 ℃ in the homogenizing process) by using a homogenizer at the rotating speed of 10000rpm until the powder reaches 200 meshes, centrifuging and filtering (the rotating speed is set to 20000 rpm), and taking a liquid phase to obtain a crude extract product. Degreasing and deodorizing the crude product of the extracting solution twice by using active carbon (each time adding active carbon accounting for 6% of the mass of the periplaneta americana medicinal material for removing, stirring for 30 min), filtering, concentrating the filtrate under reduced pressure at 70 ℃ to a small volume, and drying under reduced pressure in vacuum to obtain 18.7kg of periplaneta americana polypeptide extract.
Comparative example 1
This comparative example is substantially the same as example 1 except that no activated carbon was added thereto, and the specific procedure was as follows: 5kg of periplaneta americana medicinal material is taken, frozen to-20 ℃, and crushed into powder by a conventional medicinal material crusher (the temperature of the material is kept below 0 ℃ in the crushing process), so as to obtain a crushed material. 25L of pure water is added into the crushed materials, and the crushed materials are dispersed in the pure water (the oil layer floats on the water surface, the materials are easy to agglomerate, and the dispersibility is poor). Homogenizing and extracting (maintaining the temperature of the material at 0-10deg.C during homogenizing) with a homogenizer at 10000rpm until the powder obtained after homogenizing the crushed material reaches 200 mesh (i.e. the powder can pass through 200 mesh sieve) to obtain an extraction mixture. Centrifuging and filtering twice, wherein the first centrifuging speed is set to 8000rpm, the second centrifuging speed is set to 16000rpm, centrifuging and filtering, and collecting liquid phase to obtain crude extract. After 24 hours, no layering is found, and oil-water separation is not possible. Then adding 1kg of active carbon, stirring to form paste, and still not separating oil from water. The above experiment shows that the active carbon powder has important functions of dispersing and adsorbing grease in the process, and is a key material for preventing the extraction liquid from emulsifying and realizing the extraction of the American cockroach polypeptide by water.
Comparative example 2
Ethanol was used as the extraction solvent in this comparative example. The specific operation process is as follows: taking 5kg of periplaneta americana medicinal material raw material, and crushing the raw material into coarse powder at the temperature of below 0 ℃ to obtain periplaneta americana coarse powder. Adding 25L of ethanol solution with the volume fraction of 90% into the periplaneta americana coarse powder, and uniformly mixing to obtain a dispersion liquid. Homogenizing and extracting (maintaining the temperature of the material at 0-10deg.C during homogenizing) with a homogenizer at 10000rpm until the powder obtained after homogenizing the crushed material reaches 200 mesh (i.e. the powder can pass through 200 mesh sieve) to obtain an extraction mixture (i.e. the dispersion liquid after grinding treatment). Degreasing and deodorizing the crude product of the extracting solution twice by using active carbon (5% of active carbon of the American cockroach medicinal material mass is added each time and stirred for 30 min), and filtering to obtain a liquid phase to obtain the American cockroach polypeptide extracting solution. Concentrating the periplaneta americana polypeptide extract at 70deg.C under reduced pressure to small volume, and vacuum drying at 70deg.C under reduced pressure to obtain 1850g periplaneta americana polypeptide extract (flowable oil). The above experiments show that the active carbon can not adsorb and remove most of the grease in the ethanol extract because the ethanol is similar to the grease.
Comparative example 3
Taking the flowable oily periplaneta americana polypeptide extract in comparative example 2, adding 8 times of water, slightly stirring, standing for 24 hours, removing a floating grease layer on the liquid surface, pouring out the water solution (slightly emulsifying), adding equal amount of activated carbon (based on the amount of the extract), stirring for 30 minutes, filtering to obtain clear and transparent extract, adding equal amount of activated carbon (based on the amount of the extract), stirring for 30 minutes, filtering, concentrating the filtrate to a small volume at 70 ℃ under reduced pressure, and drying under reduced pressure in vacuum to obtain 813g of the periplaneta americana polypeptide extract. The above experiments show that the organic solvent ethanol extraction of the periplaneta americana extract can not remove most of grease through activated carbon adsorption, and after the obtained extract is added with water, the water layer containing polypeptide is easily emulsified in the dispersing and dissolving process due to the existence of a large amount of grease, and the amount of the obtained polypeptide extract is obviously lower than that of the method of the invention due to the fact that the ethanol organic solvent has low solubility to part of high molecular weight polypeptide, so that the polypeptide extraction is incomplete.
Experimental example
The periplaneta americana polypeptide extracts of examples 1 to 4 and comparative examples 1 to 3 were examined, and the examination items and examination results are shown in table 1. The detection method of the experimental example is as follows: GB/T5009.124-2003 (determination of amino acids in foods) and GB5009.5-2010 (determination of proteins in foods of national standards for food safety), data representing the mass (g/100 g) of a detection object per 100g of a detection object (periplaneta americana polypeptide extract); GB 5009.6-2016 (determination of fat in food safety national standard food), data expressed in terms of mass (g/100 g) of detection target per 100g of detection object (periplaneta americana polypeptide extract); the product yield is the mass of the periplaneta americana polypeptide extract divided by the mass of the periplaneta americana medicinal material raw material multiplied by 100%.
Table 1: experimental example detection results (N/A did not detect product)
It can be seen from table 1 that the adsorption of the activated carbon on the oil during the extraction process is the key of the present invention, so that the oil is adsorbed on the surface of the activated carbon, the phenomenon of emulsification of the extract is prevented, and the smoothness of the process is ensured (examples 1 to 4). Comparative example 1 was extracted using water as a solvent, and the active ingredient could not be extracted, resulting in failure of the extraction process. On the other hand, the existing process for extracting periplaneta americana based on organic solvents such as ethanol (comparative example 2) extracts a great amount of grease in a liquid phase while extracting functional components such as polypeptide, and the like, wherein the ethanol is similar to the grease components and is compatible with the grease components, and the purpose of adsorbing the grease components in the ethanol cannot be achieved by the activated carbon. The periplaneta americana polypeptide extract obtained in comparative example 2 is a flowable oil, and has high macroscopic lipid content, so that the composition detection is not performed. Even if the product obtained in comparative example 2 was subjected to further degreasing treatment in comparative example 3, the lipid residue of the periplaneta americana polypeptide extract obtained in comparative example 3 was still higher than that of examples 1 to 4. Comparative example 3 shows that organic solvents such as ethanol have poor solubility for polypeptides, particularly high molecular weight polypeptides, and thus the extraction is incomplete, the yield of the product is low, and the content of total proteins and total amino acids in the product is low.
The foregoing is merely exemplary embodiments of the present invention, and specific structures and features that are well known in the art are not described in detail herein. It should be noted that modifications and improvements can be made by those skilled in the art without departing from the structure of the present invention, and these should also be considered as the scope of the present invention, which does not affect the effect of the implementation of the present invention and the utility of the patent. The protection scope of the present application shall be subject to the content of the claims, and the description of the specific embodiments and the like in the specification can be used for explaining the content of the claims.
Claims (7)
1. The preparation method of the periplaneta americana polypeptide extract taking water as a medium is characterized by comprising the following steps of:
s1: mixing the periplaneta americana medicinal material with activated carbon powder, crushing at the temperature of-20-0 ℃, sieving to obtain a screen material; the mesh number of the sieve is greater than or equal to 80 meshes; the mass ratio of the periplaneta americana medicinal material raw material to the activated carbon powder is 1:1-1:2;
s2: dispersing the crushed material in water to obtain a dispersion liquid, and homogenizing the dispersion liquid to obtain an extraction mixture;
s3: and purifying and separating the periplaneta americana polypeptide extract from the extraction mixture.
2. The method according to claim 1, wherein in S2, the pulverized material is dispersed in water to obtain a dispersion, and the dispersion is subjected to a homogenization treatment until the pulverized material in the dispersion reaches 200 to 500 mesh to obtain an extraction mixture.
3. The method according to claim 2, wherein in S2, the homogenization treatment is performed in an environment of 0 to 10 ℃.
4. A method according to claim 3, wherein in S2, the water is used in an amount of: 5-10L of water is used as 1kg of periplaneta americana medicinal material raw material.
5. The process according to any one of claims 1 to 4, wherein in S3, the extract mixture is first subjected to solid-liquid separation and a liquid phase is taken to obtain a crude extract; then adding active carbon into the crude product of the extracting solution, stirring, and filtering to obtain a liquid phase to obtain a periplaneta americana polypeptide extracting solution; concentrating and drying the periplaneta americana polypeptide extract to obtain the periplaneta americana polypeptide extract.
6. The method according to claim 5, wherein in S3, the extract mixture is subjected to solid-liquid separation by centrifugation at 8000 to 25000rpm.
7. The preparation method according to claim 6, wherein in S3, the amount of activated carbon added into the crude extract is 4% -6% of the raw material mass of periplaneta americana; adding active carbon into the crude product of the extract, stirring for 30min, and filtering to obtain liquid phase to obtain the periplaneta americana polypeptide extract.
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