CN111925408A - Preparation method of periplaneta americana polypeptide extract by using water as medium - Google Patents
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- CN111925408A CN111925408A CN202010819339.XA CN202010819339A CN111925408A CN 111925408 A CN111925408 A CN 111925408A CN 202010819339 A CN202010819339 A CN 202010819339A CN 111925408 A CN111925408 A CN 111925408A
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Abstract
The invention belongs to the technical field of natural component extraction and preparation, relates to a preparation method of a periplaneta americana polypeptide extract, and particularly relates to a preparation method of a periplaneta americana polypeptide extract by taking water as a medium, which comprises the following steps in sequence: mixing the periplaneta americana medicinal material raw material with activated carbon powder, and then crushing to obtain a crushed material; dispersing the pulverized 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 residue, completely extracts polypeptide, has low extraction cost, adopts activated carbon to adsorb oil components, avoids difficult separation of the emulsification of the extracting solution, ensures the smoothness of the process, removes the fishy smell of the product through the activated carbon adsorption, improves the product quality, and has good market application prospect and potential huge economic benefit.
Description
Technical Field
The invention belongs to the technical field of natural component extraction and preparation, relates to a preparation method of a periplaneta americana polypeptide extract, and particularly relates to a preparation method of a periplaneta americana polypeptide extract by taking water as a medium.
Background
The modern research shows that the main active components of the periplaneta americana, also called blatta, are polypeptide and biological enzyme thereof, and have obvious effects of promoting tissue repair and regeneration, resisting tumors and hepatitis. The medicine on the market prepared by taking the periplaneta americana as the main raw material comprises a rehabilitation new liquid, a Xinmailong capsule, a liver dragon capsule and the like, and generates good economic and social benefits.
The periplaneta americana is an animal medicinal material, is rich in grease, and is generally prepared by degreasing an organic solvent (petroleum ether, n-hexane and the like) and then using a high-polarity solvent (alcohol or an alcohol-water mixture) as an extraction solvent to extract polypeptide from the periplaneta americana, wherein the method has the following defects: the cost of using organic solvents is high; the residue of greasy component is large; the fishy smell of the extract is not easy to remove; the low solubility of part of the polypeptide in organic solvents leads to incomplete extraction. If water is used as an extraction solvent instead of an organic solvent, although the solubility of the polypeptide in the extraction solvent can be improved, the solubility of the oil in water is extremely low, the oil content reaches a certain critical value, and the phenomenon of emulsification of an extracting solution in the extraction process is easily caused, so that target components (protein, polypeptide, amino acid and the like) cannot be effectively separated from a large amount of oil in medicinal materials, and the product quality cannot be effectively guaranteed.
Disclosure of Invention
The invention aims to provide a preparation method of a periplaneta americana polypeptide extract by taking water as a medium, which is used for solving the technical problem of emulsification when water is used as an extraction solvent.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a preparation method of a periplaneta americana polypeptide extract with water as a medium comprises the following steps in sequence:
s1: mixing the periplaneta americana medicinal material raw material with activated carbon powder, and then 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 surface of the micro powder is isolated and adsorbed, and the crushed materials are obtained after crushing treatment. The crushed material comprises active carbon powder and crushed American cockroach medicinal material raw materials. After the grinding is finished, water is used as a medium to extract the ground materials (including the activated carbon powder), the ground materials are dispersed in water to obtain a dispersion liquid, the American cockroach medicinal material is continuously ground and homogenized 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 activated carbon (the activated carbon also has the effect of assisting in grinding the medicinal material), and the polypeptide components enter the water. And grinding and extracting to obtain an extraction mixture, wherein the extraction mixture is the ground dispersion liquid. After extraction, dregs of a decoction (the dregs of a decoction contain activated carbon powder) and a liquid phase are separated from the extraction mixture, and the periplaneta americana polypeptide extract can be obtained after subsequent steps such as purification treatment and the like. In the scheme, homogenizing refers to homogenizing materials, and the materials are subjected to splitting, crushing, blending and other processes in a container by rotating and driving a rotary cutter through a motor, so that the materials are stirred and smashed. Except using motor rotation drive rotary cutter, also can use the grinder among the prior art to grind the dispersion, make the material stir and smash to pieces, also can realize the homogenate to the material. The key point of the scheme is that the active carbon powder and the animal medicinal material powder are required to exist simultaneously in the homogenizing process, and the active carbon powder plays a role in preventing system emulsification.
In the process of crushing medicinal materials and extracting polypeptide, the added active carbon powder can fully absorb grease in the medicinal materials and prevent the phenomenon of emulsification of the extracting solution, so that the efficiency of extracting the polypeptide 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 common function of the activated carbon is to remove impurities (lipids, pigments and the like) in the extract (after extraction is finished), but in the scheme, the activated carbon powder is added into the process in the process of crushing and extracting medicinal materials, so that unexpected technical effects are brought, namely the purposes of preventing the emulsification phenomenon of an extraction system and improving the product quality are achieved.
The invention provides a preparation method for extracting periplaneta americana polypeptide by using water as an extraction medium, which takes water as the medium, avoids organic solvent residue, completely extracts the polypeptide, has low extraction cost, adsorbs grease components by using activated carbon, avoids difficult separation of emulsion of an extracting solution, ensures the process smoothness, removes the fishy smell of a product by activated carbon adsorption, improves the product quality, and has better market application prospect and potential huge economic benefit.
Further, in S1, the temperature for crushing 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 polypeptide can be ensured at low temperature, and the degradation of the protein or polypeptide is prevented; the low temperature can also avoid the occurrence of the phenomenon of the decay of the periplaneta americana.
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 pulverized 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 reacts with oil substances in the processes of crushing and extracting the medicinal materials to prevent 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 and block a screen (the active carbon powder has the function of preventing the medicinal materials from blocking the screen after being crushed), and a slight emulsification phenomenon still occurs in the extraction process; and when the ratio is higher than 1:2, the effects of medicine crushing and oil absorption are not improved more remarkably, and the waste of the activated carbon material is also caused.
Further, in S1, mixing the periplaneta americana medicinal material with activated carbon powder, crushing, and sieving to obtain undersize materials; the mesh number of the sieve is greater than or equal to 80 meshes.
By adopting the technical scheme, the raw material of the periplaneta americana is crushed to be smaller than a certain particle size, so that the particles of the medicinal material can be fully contacted with medium water, and the dissolution efficiency of water on target components is improved.
Further, in S2, dispersing the pulverized material in water to obtain a dispersion, and homogenizing the dispersion until the pulverized material in the dispersion reaches 200 to 500 mesh, to obtain an extraction mixture.
By adopting the technical scheme, the homogenate is carried out until the crushed materials in the dispersion liquid reach 200-500 meshes, so that the functional components can be fully dissolved out, and the full extraction of the target components can be realized.
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 particle hardness in the temperature range, and are not softened and agglomerated, so that the homogenization efficiency is ensured; meanwhile, the liquid medicine is not frozen, so that excessive wear of equipment caused by ice particles is avoided.
Further, in S2, the amount of water used is: 5-10L of water is used for 1kg of periplaneta americana medicinal material.
By adopting the technical scheme, sufficient water can fully dissolve out functional components (polypeptide and other substances) and improve the yield of target components.
Further, in S3, first, the extraction mixture is subjected to solid-liquid separation and the liquid phase is taken out to obtain a crude extract; then adding activated carbon into the crude extract, stirring, filtering and taking a liquid phase to obtain a periplaneta americana polypeptide extract; concentrating and drying the periplaneta americana polypeptide extracting solution 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 used herein may be one conventionally used for adsorbing impurities, and the above-mentioned activated carbon powder is not necessarily used.
Further, in S3, the extraction mixture is subjected to solid-liquid separation by centrifugation at 8000 to 25000 rpm.
By adopting the technical scheme, the centrifugal rotating speed can fully separate solid from liquid, and particularly separate the active carbon powder from target components.
Further, in S3, the amount of the active carbon added into the crude extract is 4-6% of the mass of the raw materials of the periplaneta americana medicinal material; adding active carbon into the crude extract, stirring for 30min, and filtering to obtain liquid phase to obtain Periplaneta americana polypeptide extract.
By adopting the technical scheme, the using amount of the active carbon and the stirring time ensure that impurities such as grease, pigment and the like are fully removed.
Detailed Description
Example 1
Taking 5kg of periplaneta americana medicinal material, adding 5kg of activated carbon (200-300 meshes, in the embodiment, 300 meshes is specifically selected), freezing to-20 ℃, and crushing by using a crusher (keeping the temperature of the material below 0 ℃ in the crushing process) to obtain a crushed material (sieving by using a 80-mesh sieve and taking undersize materials to obtain the material). 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 at 10000rpm by a homogenizer (keeping the material temperature at 0-10 ℃ in the homogenizing process) until the powder obtained after homogenizing the crushed material reaches 200 meshes to obtain an extraction mixture (namely the dispersion liquid after homogenizing treatment). Centrifuging and filtering twice, setting the first centrifugation rotation speed at 8000rpm and the second centrifugation rotation speed at 16000rpm, centrifuging and filtering, and collecting liquid phase to obtain crude extract. And (3) degreasing and deodorizing the crude extract by using active carbon (200 meshes) twice (adding active carbon accounting for 5% of the mass of the raw material of the periplaneta americana medicinal material each time, and stirring for 30min), and filtering to obtain a liquid phase to obtain the periplaneta americana polypeptide extract. Concentrating the periplaneta americana polypeptide extract at 70 ℃ under reduced pressure to a small volume, and then drying at 70 ℃ under reduced pressure to obtain 952g periplaneta americana polypeptide extract.
Example 2
Taking 20kg of periplaneta americana medicinal material, adding 30kg of activated carbon (400-500 meshes, in the embodiment, 500 meshes is specifically selected), freezing to-10 ℃, crushing into powder by using a crusher (keeping the temperature at-5 ℃ in the crushing process), and obtaining a crushed material (sieving by using a 80-mesh sieve and taking undersize materials). Adding 200L of water into the crushed materials, uniformly mixing to obtain a dispersion, homogenizing and extracting by using a homogenizer at the rotation speed of 10000rpm (keeping the temperature of 0-10 ℃ in the homogenizing process) until the powder reaches 400 meshes to obtain an extraction mixture (namely the dispersion subjected to grinding treatment). Centrifuging and filtering (rotating speed set at 25000rpm) to extract the mixture, and collecting the liquid phase to obtain crude extract. And (3) degreasing and deodorizing the crude extract by using active carbon twice (6% of the mass of the periplaneta americana medicinal material is added each time, stirring is carried out for 30min), filtering is carried out, the filtrate is subjected to reduced pressure concentration at 70 ℃ to a small volume, and then freeze drying is carried out, so that 3660g of the periplaneta americana polypeptide extract is obtained.
Example 3
Taking 2kg of American cockroach medicinal material, adding 4kg of activated carbon (400-500 meshes, in the embodiment, 500 meshes is specifically selected), freezing to-20 ℃, grinding into powder by using a grinder (keeping the temperature below-10 ℃ in the grinding process), and obtaining a crushed material (sieving by using a 200-mesh sieve and taking undersize materials). Adding 20L of water into the crushed materials, uniformly mixing to obtain a dispersion, homogenizing and extracting by using a homogenizer at the rotating speed of 10000rpm (keeping the temperature of 0-10 ℃ in the homogenizing process) until the powder reaches 500 meshes to obtain an extraction mixture (namely the dispersion subjected to grinding treatment). Centrifuging and filtering the extract mixture twice, setting the first rotation speed at 16000rpm and the second rotation speed at 25000rpm, and collecting the liquid phase to obtain crude extract. And (3) degreasing and deodorizing the crude extract by using active carbon twice (adding active carbon accounting for 4% of the mass of the periplaneta Americana medicinal material each time for removing, stirring for 30min), filtering, concentrating the filtrate at 70 ℃ under reduced pressure to a small volume, and freeze-drying to obtain 406g of the periplaneta Americana polypeptide extract.
Example 4
Taking 100kg of periplaneta americana medicinal material, adding 100kg of activated carbon (200-300 meshes, in the embodiment, 300 meshes is specifically selected), freezing to-10 ℃, and crushing into medicinal powder (keeping the temperature below 0 ℃ in the crushing process) to obtain a crushed material (sieving by a 80-mesh sieve and taking undersize materials to obtain the powder). Adding 800L of water into the crushed materials, uniformly mixing to obtain a dispersion, performing homogenate extraction by using a homogenizer at the rotation speed of 10000rpm (keeping the temperature of 0-10 ℃ in the homogenate process) until the powder reaches 200 meshes, centrifuging and filtering (setting the rotation speed to 20000rpm), and taking a liquid phase to obtain a crude extract. And (3) degreasing and deodorizing the crude extract by using active carbon twice (6% of the mass of the periplaneta americana medicinal material is added each time, stirring is carried out for 30min), filtering is carried out, the filtrate is concentrated to a small volume at 70 ℃ under reduced pressure, and vacuum drying is carried out to obtain 18.7kg of periplaneta americana polypeptide extract.
Comparative example 1
The comparative example is basically the same as example 1, and is different in that no activated carbon is added in the comparative example, and the specific operation process is as follows: taking 5kg of Periplaneta americana medicinal material, freezing to-20 ℃, and pulverizing into powder by using a conventional medicinal material pulverizer (keeping the temperature of the material below 0 ℃ in the pulverizing process) to obtain a pulverized material. Adding 25L of pure water into the pulverized material, and dispersing the pulverized material in the pure water (oil layer floating on the water surface, the material is easy to aggregate and has poor dispersibility). Homogenizing and extracting at 10000rpm by a homogenizer (keeping the material temperature at 0-10 ℃ in the homogenizing process) until the powder obtained after the grinding material is homogenized reaches 200 meshes (namely the powder can pass through a 200-mesh sieve), and obtaining an extraction mixture. Centrifuging and filtering twice, setting the first centrifugation rotation speed at 8000rpm and the second centrifugation rotation speed at 16000rpm, centrifuging and filtering, and collecting liquid phase to obtain crude extract. Standing for 24 hr, no separation, and no oil-water separation. Then 1kg of active carbon is added and stirred to form paste, and the oil-water separation still can not be carried out. The tests show that the activated carbon powder has important functions of dispersing and adsorbing grease in the process, and is a key material for preventing the emulsification of the extracting solution and realizing the extraction of the periplaneta americana polypeptide by water.
Comparative example 2
This comparative example used ethanol as the extraction solvent. The specific operation process is as follows: 5kg of periplaneta americana medicinal material raw material is taken, and the raw material is crushed into coarse powder at the temperature of below 0 ℃ to obtain the 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 at 10000rpm by a homogenizer (keeping the material temperature at 0-10 ℃ in the homogenizing process) until the powder obtained after homogenizing the crushed material reaches 200 meshes (namely the powder can pass through a 200-mesh sieve), and obtaining an extraction mixture (namely the dispersion liquid after grinding treatment). And (3) degreasing and deodorizing the crude extract by using active carbon twice (adding active carbon accounting for 5% of the mass of the periplaneta Americana medicinal material each time, and stirring for 30min), and filtering to obtain a liquid phase to obtain the periplaneta Americana polypeptide extract. The periplaneta americana polypeptide extract is concentrated to a small volume at 70 ℃ under reduced pressure, and then dried under reduced pressure at 70 ℃ under vacuum to obtain 1850g of periplaneta americana polypeptide extract (flowable oily matter). The tests show that the active carbon can not adsorb and remove most of oil in the ethanol extract because the ethanol is similar to and compatible with the oil.
Comparative example 3
Adding 8 times of water into the oily periplaneta americana polypeptide extract in the comparative example 2, slightly stirring to disperse, standing for 24 hours, removing floating oil layers on the liquid surface, pouring out water liquid (slight emulsification), adding an equal amount of activated carbon (based on the amount of the extract), stirring for 30 minutes, filtering to obtain a clear transparent extracting solution, adding an equal amount of activated carbon (based on the amount of the extract), stirring for 30 minutes, filtering, concentrating the filtrate at 70 ℃ under reduced pressure to a small volume, and drying under reduced pressure in vacuum to obtain 813g of the periplaneta americana polypeptide extract. The experiments show that the periplaneta americana extract extracted by the ethanol as the organic solvent can not remove most of oil through activated carbon adsorption, after the extract is added with water, emulsification is easily formed in the dispersion and dissolution process due to the existence of a large amount of oil, the water layer containing the polypeptide is subjected to activated carbon adsorption treatment, the amount of the obtained polypeptide extracting solution is obviously lower than that of the method, and the incomplete extraction of the polypeptide is caused due to the low solubility of the ethanol organic solvent on part of high molecular weight polypeptide.
Examples of the experiments
The periplaneta americana polypeptide extracts of examples 1 to 4 and comparative examples 1 to 3 were tested, and the test items and the test results are shown in table 1. The detection method of this experimental example is as follows: GB/T5009.124-2003 (determination of amino acids in food) and GB5009.5-2010 (determination of proteins in food products in national standards for food safety), the data being expressed in terms of the mass (g/100g) of the test object per 100g of test object (Periplaneta americana polypeptide extract); GB 5009.6-2016 (national food safety standard for fat determination) data is expressed in the form of the mass (g/100g) of the target substance per 100g of the substance (Periplaneta americana polypeptide extract); the product yield is the product of dividing the mass of the periplaneta americana polypeptide extract by the mass of the periplaneta americana medicinal material raw material and multiplying the product by 100 percent.
Table 1: experimental example test results (N/A No product test)
As can be seen from Table 1, the key point of the present invention is to fully utilize the adsorption of the activated carbon to the oil during the extraction process, so that the oil is adsorbed on the surface of the activated carbon, the emulsification of the extraction solution is prevented, and the smoothness of the process is ensured (examples 1-4). Comparative example 1 extraction using water as a solvent failed to extract effective ingredients, resulting in failure of the extraction process. On the other hand, in the existing periplaneta americana extraction process (comparative example 2) based on organic solvents such as ethanol, a large amount of grease is also extracted into a liquid phase while functional components such as polypeptide are extracted by the ethanol, the ethanol is similar to and compatible with the grease components, and the purpose of adsorbing the grease components in the ethanol cannot be achieved by activated carbon. The periplaneta americana polypeptide extract obtained in comparative example 2 is a fluid oily substance, and the content of lipid substances is high as seen by naked eyes, so that the component detection is not performed on the periplaneta americana polypeptide extract. Even if the product obtained in comparative example 2 was subjected to further lipid removal 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 1-4. Comparative example 3 shows that organic solvents such as ethanol have poor solubility to polypeptides, especially high molecular weight polypeptides, so that the extraction is incomplete, the product yield is low, and the total protein and total amino acid content in the product is low.
The foregoing is merely an example of the present invention and common general knowledge of known specific structures and features of the embodiments is not described herein in any greater detail. It should be noted that, for those skilled in the art, without departing from the structure of the present invention, several changes and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (10)
1. A preparation method of a periplaneta americana polypeptide extract with water as a medium is characterized by comprising the following steps in sequence:
s1: mixing the periplaneta americana medicinal material raw material with activated carbon powder, and then 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.
2. The preparation method according to claim 1, wherein in S1, the temperature for crushing the mixture of the periplaneta americana medicinal material raw material and the activated carbon powder is-20-0 ℃.
3. The preparation method according to claim 2, wherein in S1, the mass ratio of the periplaneta americana medicinal material raw material to the activated carbon powder is 1: 1-1: 2.
4. The preparation method according to claim 3, wherein in S1, Periplaneta americana medicinal material is mixed with activated carbon powder, and after the mixture is crushed, the mixture is sieved to obtain undersize material; the mesh number of the sieve is greater than or equal to 80 meshes.
5. The method according to claim 4, wherein the pulverized material is dispersed in water to obtain a dispersion liquid in S2, and the dispersion liquid is homogenized until the pulverized material in the dispersion liquid reaches 200-500 mesh to obtain the extraction mixture.
6. The method according to claim 5, wherein the homogenization treatment is performed in S2 at 0-10 ℃.
7. The method according to claim 6, wherein in S2, the amount of water used is: 5-10L of water is used for 1kg of periplaneta americana medicinal material.
8. The method according to any one of claims 1 to 7, wherein in S3, the extraction mixture is first subjected to solid-liquid separation and a liquid phase is taken to obtain a crude extract; then adding activated carbon into the crude extract, stirring, filtering and taking a liquid phase to obtain a periplaneta americana polypeptide extract; concentrating and drying the periplaneta americana polypeptide extracting solution to obtain the periplaneta americana polypeptide extract.
9. The method according to claim 8, wherein the extraction mixture is subjected to solid-liquid separation by centrifugation at 8000 to 25000rpm in S3.
10. The preparation method according to claim 9, wherein in S3, the amount of activated carbon added in the crude extract is 4-6% of the raw material mass of the periplaneta americana; adding active carbon into the crude extract, stirring for 30min, and filtering to obtain liquid phase to obtain Periplaneta americana polypeptide extract.
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|---|---|---|---|---|
| CN115429819A (en) * | 2022-10-19 | 2022-12-06 | 云南中医药大学 | Method for processing and preparing high-activity leech powder |
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| CN111925408B (en) | 2023-06-06 |
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