CN110934889A - Efficient and comprehensive extraction method of effective components in amber - Google Patents
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Abstract
The invention belongs to the technical field of natural product extraction, and particularly relates to a high-efficiency comprehensive extraction method of effective components in amber. The method comprises grinding Succinum, namely grinding the sea amber of Bordeaux by using a grinder, and sieving the ground Succinum by using a sieve with more than 100 meshes to obtain Succinum powder; performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing a supercritical carbon dioxide fluid, wherein the extraction temperature is 40-80 ℃, and the extraction pressure is 30-70MPa, so as to obtain a crude amber extract; and (5) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, and distilling the eluent under reduced pressure to obtain the amber extract. The invention utilizes the supercritical carbon dioxide extraction method to extract the active ingredients in the amber under the low temperature condition, thereby preventing the problem that the active ingredients in the amber are damaged due to overhigh temperature.
Description
Technical Field
The invention belongs to the technical field of natural product extraction, and particularly relates to a high-efficiency comprehensive extraction method of effective components in amber.
Background
Amber is a mineral with values comparable to diamond, jade, and jade. Is a precious stone formed by geological movement for hundreds of millions of years on the ground bottom layer. The baltic sea amber is an amber produced along the coasts of the baltic sea and is an important source of the amber in the world, and the baltic sea amber is mostly in a faint yellow color and can be divided into a Jieper amber, a lump amber, a turbid amber, a bubble amber, a bone amber and other varieties according to the internal characteristics. In the prior art, the active ingredients in the amber are extracted by using a dry distillation method, but the extraction temperature of the dry distillation method is extremely high, and certain substances in the amber can be chemically changed under the high-temperature condition; in the prior art, a method of extracting amber at normal temperature by using ethanol exists, and although effective substances finally extracted from amber can be dissolved in ethanol, the method of directly extracting amber solid by using ethanol has extremely low efficiency, and the types of effective components which can be directly extracted are also extremely limited.
For example, the chinese invention patent application discloses an amber essence, its extraction method and application [ application No.: 201910930835.X ], the invention patent comprises the following steps: (1) preparing raw materials of amber extraction medicament: according to the mass ratio of oleic acid to purified water of 5: 3 preparing an oleic acid solution, wherein the mass ratio of stearic acid to purified water is 2: 1, preparing a stearic acid solution, wherein the mass ratio of lauric acid to purified water is 2: 1, preparing a lauric acid solution, wherein the mass ratio of propylene glycol to purified water is 3: 1, preparing a propylene glycol solution, wherein the mass ratio of the auxiliary material 1 to the purified water is 1: 1, preparing an auxiliary material 1 solution, wherein the mass ratio of the auxiliary material 2 to the purified water is 1: 1 preparing an auxiliary material solution 2; (2) preparation of amber extract: adding purified water into a stainless steel barrel, and heating to 70-80 deg.C; then adding triethanolamine, the oleic acid solution, the stearic acid solution, the lauric acid solution, the propylene glycol solution, the auxiliary material 1 solution and the auxiliary material 2 solution into the mixture by stirring to prepare an amber extracting agent; wherein the mass ratio of the purified water, the triethanolamine, the oleic acid solution, the stearic acid solution, the lauric acid solution, the propylene glycol solution, the auxiliary material 1 solution and the auxiliary material 2 solution is 10: 5: 4: 9: 9: 10: 2: 1; (3) preparing fine amber powder: cleaning the waste material of the amber leftover, drying and grinding the waste material to be below 500 meshes to obtain fine amber powder for later use; (4) extraction: under the condition of ultrasonic wave, adding the amber extraction agent into a stainless steel barrel, heating to 80-90 ℃, slowly adding the amber fine powder, keeping the ultrasonic wave and heating condition, and stirring for 1 hour to obtain an extracting solution; the mass ratio of the amber extraction medicament to the fine amber powder is 5: 1; (5) and (3) filtering: filtering the extractive solution with microporous membrane of 3.0-10.0 μm to obtain filtrate, i.e. Succinum essence.
The method provided by the invention has the advantages of simple method, good extraction effect and good product quality, but the problems are not solved yet.
Disclosure of Invention
The invention aims to solve the problems and provide a high-efficiency comprehensive extraction method for effective components in amber, which can extract the effective components in amber relatively comprehensively.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for efficiently and comprehensively extracting active ingredients from amber comprises the following steps:
the method comprises the following steps: grinding Succinum, grinding Polioephalus ambrosioides, and sieving with 100 mesh sieve to obtain Succinum powder;
step two: performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing a supercritical carbon dioxide fluid, wherein the extraction temperature is 40-80 ℃, and the extraction pressure is 30-70MPa, so as to obtain a crude amber extract;
step three: and (5) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, and distilling the eluent under reduced pressure to obtain the amber extract.
In the above method for extracting effective components from amber completely and efficiently, the flow rate of the supercritical carbon dioxide fluid in the second step is 20-30 kg/h.
In the efficient and comprehensive extraction method of the effective components in the amber, the extraction time in the second step is 1-3 h.
In the efficient and comprehensive extraction method of the effective components in the amber, the elution and separation operation of the macroporous adsorption resin column in the third step comprises the following steps:
step A: loading the column, namely loading the macroporous adsorption resin into the column by a wet method, and pretreating to remove oligomers and pore-forming agents contained in the macroporous adsorption resin;
and B: preparing a sample, namely adding the crude amber extract into absolute ethyl alcohol, and stirring until the crude amber extract is completely dissolved to obtain a sample to be purified;
and C: and D, eluting, namely adding the sample to be purified obtained in the step B into a macroporous adsorption resin column, eluting by using absolute ethyl alcohol as an eluent, and collecting the eluent.
In the efficient and comprehensive extraction method of the active ingredients in the amber, the aperture of the macroporous adsorption resin is 13-14 nm.
In the above method for extracting effective components from amber, the pretreatment in step a comprises the following steps: washing the macroporous adsorption resin with 95% ethanol at a flow rate of 2BV/h until the effluent is clear and transparent, washing with water at a flow rate of 2BV/h for 1-3h, washing with a hydrochloric acid solution with a mass fraction of 4% at a flow rate of 4BV/h, and finally washing away the hydrochloric acid with water until the pH value of the effluent is 6-8.
In the efficient and comprehensive extraction method of the active ingredients in the amber, the mass of the absolute ethyl alcohol for dissolving the crude amber extract in the step B is 2-3 times of that of the crude amber extract.
In the efficient and comprehensive extraction method of the active ingredients in the amber, the height of the macroporous adsorption resin column in the step C is more than 10 times of the inner diameter of the macroporous adsorption resin column.
In the method for efficiently and comprehensively extracting the active ingredients from the amber, the vacuum degree of the reduced pressure distillation in the step three is 40-60Pa, the temperature is 100-120 ℃, and the reduced pressure distillation time is 10-20 min.
Compared with the prior art, the invention has the advantages that:
1. the invention utilizes the supercritical carbon dioxide extraction method to extract the active ingredients in the amber under the low temperature condition, thereby preventing the problem that the active ingredients in the amber are damaged due to overhigh temperature.
2. According to the invention, the ambrosia is ground into powder with a fine particle size before extraction, so that the extraction efficiency is improved, and the comprehensiveness of the extracted effective components is improved.
3. The method has the advantages of simple operation method and high extraction efficiency, and is suitable for large-scale popularization and application.
Drawings
FIG. 1 is a GC-MS spectrum of the amber extract obtained in application example 1;
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and specific embodiments.
Example 1
The embodiment provides a method for efficiently and comprehensively extracting active ingredients from amber, which comprises the following steps:
the method comprises the following steps: grinding the amber, namely grinding the bolate sea amber by using a grinder, and sieving the ground bolate sea amber by using a 200-mesh sieve to obtain amber powder;
step two: performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing a supercritical carbon dioxide fluid, wherein the extraction temperature is 80 ℃, the extraction pressure is 70MPa, the flow rate of the supercritical carbon dioxide fluid is 30kg/h, and the extraction time is 3h to obtain a crude amber extract;
step three: and (3) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, wherein the eluting and separating operation comprises the following steps:
step A: loading the macroporous adsorption resin with the aperture of 14nm into a column by a wet method, washing the macroporous adsorption resin by 95% ethanol at the flow rate of 2BV/h until the effluent liquid is clear and transparent, then washing for 3h by water at the flow rate of 2BV/h, then washing by a hydrochloric acid solution with the mass fraction of 4% at the flow rate of 4BV/h, and finally washing away the hydrochloric acid by water until the pH value of the effluent liquid is 8 so as to remove oligomers and pore-forming agents contained in the macroporous adsorption resin;
and B: preparing a sample, namely adding the crude amber extract into absolute ethyl alcohol with the mass being 3 times of that of the crude amber extract, and stirring until the crude amber extract is completely dissolved to obtain a sample to be purified;
and C: eluting, namely adding the sample to be purified obtained in the step B into a macroporous adsorption resin column, wherein the column height of the macroporous adsorption resin column is 15 times of the inner diameter of the macroporous adsorption resin column, eluting by using absolute ethyl alcohol as an eluent, and collecting the eluent;
distilling the eluate under reduced pressure to obtain amber extract, wherein the vacuum degree of the reduced pressure distillation is 60Pa, the temperature is 120 ℃, and the reduced pressure distillation time is 20 min.
The amber with different origin areas has slightly different components, and screening shows that the ambrosia parvifolia can extract more comprehensive effective components by using the method.
The amber is crushed to a smaller particle size before extraction, the extraction effect is improved, and the comprehensiveness of the extraction of effective components can be improved at the same time, when the particle size of the amber powder is larger, for example, more than 200 microns, a plurality of effective components such as isopropyl benzene (see table 1, components with the retention time of 6.912 min), d-camphor (see table 1, components with the retention time of 10.877 min), 2, 6-dimethyl tetrazine (see table 1, components with the retention time of 13.849 min) and the like cannot be extracted after extraction, and the three components are only taken as an example and do not represent the difference.
By using the supercritical carbon dioxide extraction method, the active ingredients in the amber are extracted under the low temperature condition, so that the problem that the active ingredients in the amber are damaged due to overhigh temperature can be prevented.
Preferably, the gas-liquid separation can be realized by using a flash tank in the process of reduced pressure distillation, so that the load and loss of the heating furnace are obviously reduced, the heating efficiency is improved, and the energy-saving effect is good.
Example 2
The embodiment provides a method for efficiently and comprehensively extracting active ingredients from amber, which comprises the following steps:
the method comprises the following steps: grinding the amber, namely grinding the bolate sea amber by using a grinder, and sieving the ground bolate sea amber by using a 150-mesh sieve to obtain amber powder;
step two: performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing supercritical carbon dioxide fluid, wherein the extraction temperature is 40 ℃, the extraction pressure is 30MPa, the flow rate of the supercritical carbon dioxide fluid is 20kg/h, and the extraction time is 1h to obtain a crude amber extract;
step three: and (3) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, wherein the eluting and separating operation comprises the following steps:
step A: loading a column, namely loading the macroporous adsorption resin with the aperture of 13nm into the column by a wet method, washing the macroporous adsorption resin by 95% ethanol at the flow rate of 2BV/h until an effluent liquid is clear and transparent, then washing for 1h by water at the flow rate of 2BV/h, then washing by a hydrochloric acid solution with the mass fraction of 4% at the flow rate of 4BV/h, and finally washing away hydrochloric acid by water until the pH value of the effluent liquid is 6 to remove oligomers and pore-forming agents contained in the macroporous adsorption resin;
and B: preparing a sample, namely adding the crude amber extract into absolute ethyl alcohol with the mass 2 times of that of the crude amber extract, and stirring until the crude amber extract is completely dissolved to obtain a sample to be purified;
and C: eluting, namely adding the sample to be purified obtained in the step B into a macroporous adsorption resin column, wherein the column height of the macroporous adsorption resin column is 12 times of the inner diameter of the macroporous adsorption resin column, eluting by using absolute ethyl alcohol as an eluent, and collecting the eluent;
distilling the eluate under reduced pressure to obtain amber extract, wherein the vacuum degree of the reduced pressure distillation is 40Pa, the temperature is 100 ℃, and the reduced pressure distillation time is 10 min.
Example 3
The embodiment provides a method for efficiently and comprehensively extracting active ingredients from amber, which comprises the following steps:
the method comprises the following steps: grinding the amber, namely grinding the bolate sea amber by using a grinder, and sieving the ground bolate sea amber by using a 100-mesh sieve to obtain amber powder;
step two: performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing a supercritical carbon dioxide fluid, wherein the extraction temperature is 60 ℃, the extraction pressure is 50MPa, the flow rate of the supercritical carbon dioxide fluid is 25kg/h, and the extraction time is 2h to obtain a crude amber extract;
step three: and (3) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, wherein the eluting and separating operation comprises the following steps:
step A: loading a column, namely loading the macroporous adsorption resin with the aperture of 13nm into the column by a wet method, washing the macroporous adsorption resin by 95% ethanol at the flow rate of 2BV/h until an effluent liquid is clear and transparent, then washing for 2h by water at the flow rate of 2BV/h, then washing by a hydrochloric acid solution with the mass fraction of 4% at the flow rate of 4BV/h, and finally washing away hydrochloric acid by water until the pH value of the effluent liquid is 7 so as to remove oligomers and pore-forming agents contained in the macroporous adsorption resin;
and B: preparing a sample, namely adding the crude amber extract into absolute ethyl alcohol with the mass being 2.5 times of that of the crude amber extract, and stirring until the crude amber extract is completely dissolved to obtain a sample to be purified;
and C: eluting, namely adding the sample to be purified obtained in the step B into a macroporous adsorption resin column, wherein the column height of the macroporous adsorption resin column is 10 times of the inner diameter of the macroporous adsorption resin column, eluting by using absolute ethyl alcohol as an eluent, and collecting the eluent;
distilling the eluate under reduced pressure to obtain amber extract, wherein the vacuum degree of the reduced pressure distillation is 50Pa, the temperature is 110 ℃, and the reduced pressure distillation time is 15 min.
Application example 1
20kg of Porphyra tenera amber was weighed, an amber extract was obtained by the method described in example 3, and the components were analyzed by GC, and the obtained spectrum was shown in FIG. 1, and the compounds corresponding to the respective peak positions were shown in Table 1:
TABLE 1
And (4) analyzing results: as can be seen from the above table, the extraction method provided by the present invention can efficiently and comprehensively extract the effective components in amber, thereby achieving the intended purpose of the present invention.
The specific embodiments described herein are merely illustrative of the spirit of the invention. Various modifications or additions may be made to the described embodiments or alternatives may be employed by those skilled in the art without departing from the spirit or ambit of the invention as defined in the appended claims.
Claims (9)
1. A method for efficiently and comprehensively extracting active ingredients from amber is characterized by comprising the following steps:
the method comprises the following steps: grinding Succinum, grinding Polioephalus ambrosioides, and sieving with 100 mesh sieve to obtain Succinum powder;
step two: performing supercritical carbon dioxide extraction, namely adding the amber powder obtained in the step one into an extraction kettle of a supercritical extraction device, and then introducing a supercritical carbon dioxide fluid, wherein the extraction temperature is 40-80 ℃, and the extraction pressure is 30-70MPa, so as to obtain a crude amber extract;
step three: and (5) refining, namely eluting and separating the crude amber extract obtained in the step two by using a macroporous adsorption resin column, and distilling the eluent under reduced pressure to obtain the amber extract.
2. The method for efficiently and comprehensively extracting active ingredients from amber as claimed in claim 1, wherein the method comprises the following steps: the flow rate of the supercritical carbon dioxide fluid in the second step is 20-30 kg/h.
3. The method for efficiently and comprehensively extracting active ingredients from amber as claimed in claim 1, wherein the method comprises the following steps: the extraction time in the second step is 1-3 h.
4. The method for efficiently and comprehensively extracting active ingredients from amber as claimed in claim 1, wherein the method comprises the following steps: the elution and separation operation of the macroporous adsorption resin column in the third step comprises the following steps:
step A: loading the column, namely loading the macroporous adsorption resin into the column by a wet method, and pretreating to remove oligomers and pore-forming agents contained in the macroporous adsorption resin;
and B: preparing a sample, namely adding the crude amber extract into absolute ethyl alcohol, and stirring until the crude amber extract is completely dissolved to obtain a sample to be purified;
and C: and D, eluting, namely adding the sample to be purified obtained in the step B into a macroporous adsorption resin column, eluting by using absolute ethyl alcohol as an eluent, and collecting the eluent.
5. The method for extracting effective components from amber as claimed in claim 4, wherein the method comprises the following steps: the pore diameter of the macroporous adsorption resin is 13-14 nm.
6. The method for extracting effective components from amber as claimed in claim 4, wherein the method comprises the following steps: the pretreatment in the step A comprises the following operations: washing the macroporous adsorption resin with 95% ethanol at a flow rate of 2BV/h until the effluent is clear and transparent, washing with water at a flow rate of 2BV/h for 1-3h, washing with a hydrochloric acid solution with a mass fraction of 4% at a flow rate of 4BV/h, and finally washing away the hydrochloric acid with water until the pH value of the effluent is 6-8.
7. The method for extracting effective components from amber as claimed in claim 4, wherein the method comprises the following steps: the mass of the absolute ethyl alcohol for dissolving the crude amber extract in the step B is 2-3 times of that of the crude amber extract.
8. The method for extracting effective components from amber as claimed in claim 4, wherein the method comprises the following steps: and C, the height of the macroporous adsorption resin column in the step C is more than 10 times of the inner diameter of the macroporous adsorption resin column.
9. The method for efficiently and comprehensively extracting active ingredients from amber as claimed in claim 1, wherein the method comprises the following steps: the vacuum degree of the reduced pressure distillation in the third step is 40-60Pa, the temperature is 100-120 ℃, and the reduced pressure distillation time is 10-20 min.
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