CN115557869A - Deep eutectic solvent and method for extracting astaxanthin - Google Patents
Deep eutectic solvent and method for extracting astaxanthin Download PDFInfo
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Abstract
Embodiments of the present application provide a deep eutectic solvent comprising at least two of DL-menthol, thymol, camphor, and alpha-terpineol, and a method of extracting astaxanthin. The deep eutectic solvent provided by the embodiment of the application contains terpenoid compounds, realizes the high-efficiency extraction of astaxanthin by utilizing a similar compatibility principle, has the advantages of high extraction efficiency, no toxicity, small damage to the astaxanthin structure, environmental friendliness and the like, is simple and safe to operate, has low cost, and is suitable for industrial mass production.
Description
Technical Field
The application relates to the technical field of natural product extraction, in particular to a deep eutectic solvent and a method for extracting astaxanthin.
Background
Astaxanthin is an important secondary carotenoid, has extremely strong oxidation resistance and excellent coloring function, and is widely applied to the aspects of food, chemical industry, medical treatment, aquaculture and the like. The chemical synthesis and natural extraction are two ways for obtaining astaxanthin, wherein the antioxidant activity of natural astaxanthin is obviously higher than that of synthetic astaxanthin, and the natural astaxanthin can be combined with protein or lipid and has better stability, so that the natural extraction is a main effective way for obtaining astaxanthin.
In the prior art, organic solvents such as dichloromethane, acetone and ethanol are mostly adopted for extracting astaxanthin, but the dosage of the reagent is large, the use of volatile toxic reagents such as acetone has great potential safety and health hazards, the green chemistry principle is not met, and some organic solvents can damage the structure of the astaxanthin, so that the activity of the astaxanthin is reduced, and the stability is reduced.
Therefore, there is a need in the art to provide a green substitute for astaxanthin extraction that can replace traditional organic solvents.
Disclosure of Invention
It is an object of the present application to provide a green and efficient method for extracting astaxanthin from an astaxanthin source material, in particular from haematococcus pluvialis. The above object of the present application is at least achieved by providing a deep eutectic solvent.
Objects of the present application are not limited to the above objects, and other objects and advantages of the present application, which are not mentioned above, can be understood from the following description and more clearly understood through embodiments of the present application. Further, it is easily understood that the objects and advantages of the present application can be achieved by the features disclosed in the claims and the combinations thereof.
In order to achieve the above object, the following technical solutions are adopted in the present application:
the present application first provides a natural deep eutectic solvent comprising terpenoids, the solvent comprising at least two of DL-menthol, thymol, camphor and alpha-terpineol.
In some of these embodiments, the deep eutectic solvent comprises DL-menthol and thymol, or DL-menthol and camphor, or DL-menthol and alpha-terpineol, or thymol and camphor, or thymol and alpha-terpineol, or camphor and alpha-terpineol.
In some of these embodiments, the deep eutectic solvent comprises DL-menthol and thymol in a molar ratio of 1.
In some of these embodiments, the deep eutectic solvent is in a liquid state at room temperature.
The preparation method of the deep eutectic solvent comprises the following steps:
uniformly mixing the raw materials according to the preset dosage at 70-90 ℃ so as to form the liquid deep eutectic solvent.
The application also provides the use of the deep eutectic solvent as described above in the extraction of astaxanthin.
The application also provides a method for extracting astaxanthin, which comprises the following steps:
mixing raw materials including astaxanthin source materials and the deep eutectic solvent so as to obtain an astaxanthin extracting solution.
In some embodiments, the solid-to-liquid ratio of the astaxanthin source material to the deep eutectic solvent is 1.
In some embodiments, the raw materials including the astaxanthin source material and the deep eutectic solvent are mixed and then subjected to a shaking treatment;
optionally, the oscillation speed is 800 to 1200rmp, preferably 1200rmp, and the time is 0.5 to 3 hours;
optionally, the shaking treatment is performed at a temperature of 40 to 80 ℃.
In some of these embodiments, the astaxanthin source material comprises Haematococcus pluvialis.
Optionally, the haematococcus pluvialis is subjected to cell disruption treatment before being mixed with the deep eutectic solvent;
optionally, hydrochloric acid is adopted for cell disruption treatment, the concentration of the hydrochloric acid is 2-5M, the mass ratio of haematococcus pluvialis to the hydrochloric acid is 1 (5-15), the treatment time is 2min, and the treatment temperature is 50-80 ℃.
The technical scheme provided by the embodiment of the application can have the following beneficial effects:
the deep eutectic solvent provided by the embodiment of the application contains terpenoid compounds, realizes the high-efficiency extraction of astaxanthin by utilizing a similar compatibility principle, has the advantages of high extraction efficiency, no toxicity, small damage to the astaxanthin structure, environmental friendliness and the like, is simple and safe to operate, has low cost, and is suitable for industrial mass production.
Drawings
Other features, objects and advantages of the present application will become more apparent upon reading of the following detailed description of non-limiting embodiments thereof, made with reference to the accompanying drawings in which:
FIG. 1 is a graph showing the results of the extraction yields of astaxanthin from deep eutectic solvents obtained by mixing DL-menthol and thymol at different molar ratios.
FIG. 2 is a graph of the results of various solvent extractions of astaxanthin on DPPH radical scavenging.
Detailed Description
The present application will be described in further detail with reference to examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not to be construed as limiting the invention.
It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict.
It is noted that the endpoints of the ranges and any values disclosed herein are not limited to the precise range or value, and that such ranges or values are understood to encompass values close to those ranges or values. For ranges of values, between the endpoints of each of the ranges and the individual points, and between the individual points may be combined with each other to give one or more new ranges of values, and these ranges of values should be considered as specifically disclosed herein.
The examples, where specific techniques or conditions are not indicated, are to be construed according to the techniques or conditions described in the literature in the art or according to the product specifications. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.
In the description of the present specification, reference to the description of "one embodiment," "some embodiments," "an example," "a specific example," or "some examples" or the like means that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present application. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Furthermore, various embodiments or examples and features of different embodiments or examples described in this specification can be combined and combined by one skilled in the art without contradiction.
I. Term(s)
The term "deep eutectic solvent" is also called a eutectic solvent, and is a eutectic mixture formed by two or more components through Hydrogen bonding and in a liquid state at normal temperature, and the components of the eutectic mixture include a Hydrogen Bond Acceptor (HBA) and a Hydrogen Bond Donor (HBD). The eutectic solvent may be formed by any method known in the art. For example, a compound that is solid at 25 ℃ and in a solid state (e.g., in a powder state) can be mixed directly, then heated slightly to melt and fully homogenize the solid, followed by cooling, and the resulting eutectic solvent remains in a liquid state at 25 ℃. Additional liquid ingredients (e.g., water or glycerol) may be added to the aforementioned compounds to promote eutectic formation.
The term "Haematococcus pluvialis" as used herein refers to a freshwater species of the phylum Chlorophyceae, which belongs to the class Chlorophyceae.
The term "cell disruption" as used herein refers to a process of disrupting the cellular integrity of H.pluvialis, and in particular to disrupting the cell wall of H.pluvialis such that the components within it are released, for example astaxanthin stored within the cell.
II. Detailed description of the preferred embodiments
As mentioned above, the existing organic solvents for extracting astaxanthin have the disadvantages of high toxicity, high use cost, influence on the stability and activity of astaxanthin and the like. In view of this, the present inventors have conducted extensive studies on a solvent which can be used as an astaxanthin extractant and an extraction technique thereof.
The research of the application discovers that the deep eutectic solvent for extracting the astaxanthin can be formed by mixing at least two of DL-menthol, thymol, camphor and alpha-terpineol according to a certain stoichiometric ratio, and the deep eutectic solvent for extracting the astaxanthin has the advantages of high extraction efficiency, no toxicity, lower use cost, good selectivity, high separation efficiency and capability of recycling and reusing. The present application has been completed based on the above findings.
In a first aspect of the present application, the present application provides a deep eutectic solvent comprising at least two of DL-menthol, thymol, camphor and alpha-terpineol, for example comprising DL-menthol and thymol, or comprising DL-menthol and camphor, or comprising DL-menthol and alpha-terpineol, or comprising thymol and camphor, or comprising thymol and alpha-terpineol, or comprising camphor and alpha-terpineol, or comprising DL-menthol, thymol and camphor, or comprising DL-menthol, thymol and alpha-terpineol, or comprising DL-menthol, camphor and alpha-terpineol, or comprising thymol, camphor and alpha-terpineol, or comprising DL-menthol, thymol, camphor and alpha-terpineol.
In the above embodiments of the present application, a component may act as a hydrogen bond acceptor in one specific combination and a hydrogen bond donor in another specific combination, or may act as a hydrogen bond acceptor donor with another component in one specific combination. Illustratively, when the deep eutectic solvent comprises DL-menthol and thymol, the DL-menthol is a hydrogen bond acceptor and the thymol is a hydrogen bond donor; when DL-menthol and camphor are included, DL-menthol is a hydrogen bond donor and camphor is a hydrogen bond acceptor; when DL-menthol and alpha-terpineol are included, they are hydrogen bonded to each other to provide an acceptor; when thymol and camphor are included, thymol is a hydrogen bond donor and camphor is a hydrogen bond acceptor; when thymol and alpha-terpineol are included, thymol is a hydrogen bond donor and alpha-terpineol is a hydrogen bond acceptor.
The components are selected from natural components and can be used as additives of foods, cosmetics and the like, the prepared natural deep eutectic solvent is non-toxic, has similar chemical properties to natural astaxanthin, can achieve better extraction effect, has small damage to the structure of the astaxanthin, and can replace the traditional organic solvent to be used as a novel green extraction agent of the astaxanthin. The extraction method using the solvent is safer, can maintain the activity of the astaxanthin to the greatest extent, and relieves the defects that the solvent needs to be removed after the extraction by using the traditional solvent, the process is complicated, the time consumption is long, and the method is not environment-friendly.
Further, in some preferred embodiments herein, the deep eutectic solvent comprises DL-menthol and thymol, or DL-menthol and camphor, or DL-menthol and alpha-terpineol, or thymol and camphor, or thymol and alpha-terpineol, or camphor and alpha-terpineol.
Further, in some preferred embodiments herein, the deep eutectic solvent comprises DL-menthol and thymol in a molar ratio of 1.
The molar ratio range of the hydrogen bond donor and the hydrogen bond donor is optimized, more appropriate solvent viscosity can be obtained, the formed hydrogen bond is stable, and higher extraction efficiency is obtained.
Further, in some preferred embodiments of the present application, the deep eutectic solvent is in a liquid state at room temperature. Wherein, the room temperature is 20-35 ℃.
Further, in some preferred embodiments of the present application, the deep eutectic solvent is prepared as follows:
uniformly mixing the raw materials according to the preset dosage at 70-90 ℃ so as to form the liquid deep eutectic solvent.
In a second aspect of the present application, there is provided the use of a deep eutectic solvent as described above in the extraction of astaxanthin, as an extractant for the extraction of astaxanthin from astaxanthin-rich materials, such as shrimp crab shells, haematococcus pluvialis, phaffia rhodozyma and the like.
In a third aspect of the present application, there is provided a method for extracting astaxanthin comprising the steps of:
mixing raw materials including astaxanthin source materials and the deep eutectic solvent so as to obtain an astaxanthin extracting solution.
The astaxanthin-derived material refers to a material rich in natural astaxanthin, and may be, for example, shells of shrimps and crabs, haematococcus pluvialis, phaffia rhodozyma, etc.
The mixing is not particularly limited in this application and may be carried out by procedures well known to those skilled in the art.
Further, in some preferred embodiments of the present application, the solid-to-liquid ratio of the astaxanthin source material and the deep eutectic solvent is 1.
Typical but non-limiting solid-to-liquid ratios of astaxanthin source material to deep eutectic solvent are, for example, 1.
The proportion of astaxanthin source materials and the deep eutectic solvent is optimized, the better extraction efficiency can be achieved, the extraction is more thorough, and the cost of the materials serving as raw materials is saved.
Further, in some preferred embodiments of the present application, the raw materials including the astaxanthin source material and the deep eutectic solvent are mixed and then subjected to a shaking process;
optionally, the oscillation speed is 800-1200 rmp, and the time is 0.5-3 h;
optionally, the shaking treatment is performed at a temperature of 40 to 80 ℃.
Typical but non-limiting oscillation speeds are for example 800rmp, 900rmp, 1000rmp, 1100rmp, 1200rmp or any value in between two adjacent values. Typical but non-limiting oscillation times are for example: 0.5h, 0.8h, 1h, 1.2h, 1.5h, 2h, 2.5h, 3h or any value between two adjacent values; typical but non-limiting process temperatures are for example: 40 ℃, 45 ℃, 50 ℃, 55 ℃, 60 ℃, 65 ℃, 70 ℃, 75 ℃, 80 ℃ or any value between two adjacent values.
The vibration speed, time and temperature are optimized, the astaxanthin in the astaxanthin source material can be fully separated from the material and enter the deep eutectic solvent, and a better extraction effect and a better separation effect are achieved.
Further, in some preferred embodiments herein, the astaxanthin source material comprises Haematococcus pluvialis; optionally, the haematococcus pluvialis is subjected to a cell disruption treatment prior to mixing with the deep eutectic solvent.
The commonly used method for breaking the cell wall of haematococcus pluvialis comprises mechanical wall breaking, biological enzymatic wall breaking, chemical wall breaking and the like. The mechanical wall breaking method comprises grinding method, high pressure homogenizing method, ultrasonic method, freeze thawing temperature difference method, etc. The biological enzymolysis wall breaking comprises treatment with helicase, cellulase, and pectinase. Chemical wall breaking includes acid hydrolysis and alkaline hydrolysis, for example by treatment with lactic acid, hydrochloric acid or acetic acid. It is understood that the wall breaking can also be performed using emerging technologies such as pulsed electric field, high voltage microfluidization, ionic liquids, etc.
Further, in some preferred embodiments of the present application, hydrochloric acid is used for cell disruption treatment, the concentration of the hydrochloric acid is 2-5M, the mass ratio of haematococcus pluvialis to the hydrochloric acid is 1 (5-15), the treatment time is 2min, and the treatment temperature is 50-80 ℃. Preferably, the concentration of the hydrochloric acid is 4M, the mass ratio of the haematococcus pluvialis to the hydrochloric acid is 1.
The concentration of hydrochloric acid, the mass ratio of haematococcus pluvialis to the hydrochloric acid, the treatment time and the temperature are optimized, the cell wall of the haematococcus pluvialis can be fully broken, and a better extraction effect is achieved.
Further, in some preferred embodiments of the present application, the haematococcus pluvialis is subjected to cell disruption treatment with hydrochloric acid, the supernatant is removed by centrifugation, the precipitate is washed with deionized water to remove the acid, and the washed product is freeze-dried.
Further, in some preferred embodiments of the present application, after the shaking treatment, solid-liquid separation is performed to obtain the astaxanthin extraction solution.
The solid-liquid separation is not particularly limited, and is selected from any one of filtration, centrifugal separation, gravity settling and centrifugal settling, and centrifugal separation is preferred. The rotational speed of the centrifugal separation is preferably 2000 to 8000rmp, more preferably 5000rmp, and the time of the centrifugal separation is preferably 2 to 15min, more preferably 5min.
Further, in some preferred embodiments of the present application, the method further comprises a step of separating and purifying astaxanthin from the astaxanthin extracting solution.
The technical effects provided by the present invention will be described in detail with reference to the following examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Step 1): mixing DL-menthol and thymol according to a molar ratio of 1.
Step 2): mixing haematococcus pluvialis with hydrochloric acid according to a mass ratio of 1.
Step 3): and (3) mixing the wall-broken haematococcus pluvialis and the deep eutectic solvent according to the solid-liquid ratio of 1: mixing at 80g/mL, oscillating at 60 deg.C with metal bath at controlled oscillation speed of 1200r/min for 2 hr, centrifuging at 5000rmp for 5min, and separating solid and liquid to obtain astaxanthin extractive solution.
The astaxanthin content in the astaxanthin extracting solution is measured by a spectrophotometry. The extraction yield = (C × V)/M, wherein C is the concentration of astaxanthin in the astaxanthin extracting solution, and mg/mL; v is the volume of the astaxanthin extracting solution, mL; m is the mass of haematococcus pluvialis, g.
Repeating for three times, and calculating to obtain the extraction yield of 41.70 +/-0.4 mg/g.
Example 2
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and camphor were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 24.77 +/-1.8 mg/g.
Example 3
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and α -terpineol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 33.35 +/-0.6 mg/g.
Example 4
An astaxanthin extract was obtained by referring to the method of example 1 except that, in step 1), thymol and camphor were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 35.65 +/-2.8 mg/g.
Example 5
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), thymol and α -terpineol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 39.56 +/-2.3 mg/g.
Example 6
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), camphor and α -terpineol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 26.70 +/-2.1 mg/g.
Example 7
Referring to the method of example 1, except that, in step 3), the wall-broken haematococcus pluvialis and deep eutectic solvent are mixed according to a solid-to-liquid ratio of 1: mixing at 60g/mL, performing shaking treatment at 60 deg.C with metal bath at controlled shaking speed of 1200r/min for 2 hr, centrifuging at 5000rmp speed for 5min, and performing solid-liquid separation to obtain astaxanthin extractive solution.
Repeating for three times, and calculating to obtain the extraction yield of 42.22 +/-1.7 mg/g.
Example 8
Referring to the method of example 1, except that, in step 3), the wall-broken haematococcus pluvialis and the deep eutectic solvent are mixed according to a solid-to-liquid ratio of 1: mixing at 60g/mL, shaking at 70 deg.C with metal bath at controlled shaking speed of 1200r/min for 2 hr, centrifuging at 5000rmp for 5min, and separating solid and liquid to obtain astaxanthin extractive solution.
Repeating for three times, and calculating to obtain the extraction yield of 43.31 +/-0.4 mg/g.
Example 9
An astaxanthin extract was obtained by referring to the method of example 1, except that,
in the step 1), mixing DL-menthol and thymol according to a molar ratio of 1.
In the step 3), the haematococcus pluvialis subjected to wall breaking and the deep eutectic solvent are mixed according to the solid-to-liquid ratio of 1: mixing at 20g/mL, oscillating at 40 deg.C for 0.5 hr with metal bath at controlled oscillation speed of 800r/min, centrifuging at 5000rmp for 5min, and separating solid and liquid to obtain astaxanthin extractive solution.
Repeating for three times, and calculating to obtain the extraction yield of 40.15 +/-0.95 mg/g.
Example 10
An astaxanthin extract was obtained by referring to the method of example 1, except that,
in the step 1), mixing DL-menthol and thymol according to a molar ratio of 1.
In the step 3), the wall-broken haematococcus pluvialis and the deep eutectic solvent are mixed according to the solid-liquid ratio of 1: mixing at a ratio of 100g/mL, shaking at 80 deg.C with metal bath at controlled shaking speed of 1000r/min for 3 hr, centrifuging at 5000rmp for 5min, and separating solid and liquid to obtain astaxanthin extractive solution.
Repeating for three times, and calculating to obtain the extraction yield of 45.60 +/-1.87 mg/g.
Example 11
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 32.80 +/-2.95 mg/g.
Example 12
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 33.95 +/-0.62 mg/g.
Example 13
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 1.
Repeating for three times, and calculating to obtain the extraction yield of 33.08 +/-2.16 mg/g.
Example 14
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 1.5, shaken in a constant temperature metal bath at 80 ℃ to be clear and transparent, cooled and left to stand to room temperature to obtain a deep eutectic solvent.
Repeating for three times, and calculating to obtain the extraction yield of 34.60 +/-0.88 mg/g.
Example 15
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 1.5.
Repeating for three times, and calculating to obtain the extraction yield of 35.28 +/-0.96 mg/g.
Example 16
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed at a molar ratio of 2.
Repeating for three times, and calculating to obtain the extraction yield of 32.94 +/-1.83 mg/g.
Example 17
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed in a molar ratio of 3.
Repeating for three times, and calculating to obtain the extraction yield of 31.78 +/-2.11 mg/g.
Example 18
An astaxanthin extract was obtained by referring to the method of example 1, except that, in the step 1), DL-menthol and thymol were mixed at a molar ratio of 4.
Repeating for three times, and calculating to obtain the extraction yield of 29.78 +/-1.41 mg/g.
In the above examples, the extraction yield of astaxanthin from deep eutectic solvents obtained by mixing DL-menthol and thymol in different molar ratios is shown in fig. 1, and it can be seen that the extraction yield tends to increase and decrease with the increase of the molar ratio of DL-menthol to thymol, and the extraction effect is the best with a molar ratio of 1.
Comparative example 1
An astaxanthin extract was obtained by referring to the method of example 1 except that ethanol was used as an extractant. Repeating for three times, and calculating to obtain the extraction yield of 35.01 +/-1.40 mg/g.
Comparative example 2
An astaxanthin extract was obtained by referring to the method of example 1, except that caprylic/capric triglyceride (GTCC) was used as an extractant. Repeating for three times, and calculating to obtain the extraction yield of 44.08 +/-1.10 mg/g.
Comparative example 3
An astaxanthin extract was obtained by referring to the method of example 1, except that acetone was used as an extractant. Repeating for three times, and calculating to obtain the extraction yield of 40.43 +/-0.93 mg/g.
In conclusion, the deep eutectic solvent provided by the application has an excellent extraction effect on astaxanthin, the effect is equivalent to that of a common organic solvent and even superior to that of ethanol, the deep eutectic solvent can be used as a green solvent for extracting astaxanthin, and the deep eutectic solvent is non-toxic and simple in process.
Test example DPPH radical scavenging test
Weighing 4mg DPPH, diluting to 50mL with ethanol to obtain a solution with a concentration of 2X 10 -4 A solution of DPPH in mol/L. Then, the extract was diluted with ethanol to an appropriate concentration, and the extracts obtained in example 1 and comparative examples 1 to 3 were each diluted by the same factor. And sequentially adding 100 mu L of LDPPH solution and 100 mu L of diluted extracting solution into a 96-well plate, carrying out dark reaction for 30min at normal temperature, and then measuring the light absorption value Ax at the wavelength of 517nm by using a microplate reader. The absorbance A0 of the diluted extract was measured by using 100. Mu.L of the extract instead of the test solution, and the absorbance A1 of the diluted extract was measured by using 100. Mu.L of ethanol instead of DPPH. Each concentration measurement was performed 3 times in parallel and the clearance was calculated according to the formula:
and the clearance of the extract after being stored for 4, 5, 7, 8, 10, 11, 12, 13 and 15 days at 40 ℃ in a dark place is respectively measured. The results of the experiment are shown in FIG. 2.
Through calculation, the DPPH free radical clearance rate of the extracting solution obtained in the example 1 is 98.1%, the DPPH free radical clearance rate of the extracting solution obtained in the comparative example 1 is 76.7%, the DPPH free radical clearance rate of the extracting solution obtained in the comparative example 2 is 89.2%, and the DPPH free radical clearance rate of the extracting solution obtained in the comparative example 3 is 80.9%, which shows that the astaxanthin activity of the astaxanthin extracted by the deep eutectic solvent is high, and the antioxidant activity of the astaxanthin can be better reserved by the deep eutectic solvent. Further, as can be seen from fig. 2, the astaxanthin activity stability extracted by the deep eutectic solvent of the present application is high.
The above description is only a preferred embodiment of the application and is illustrative of the principles of the technology employed. It will be appreciated by those skilled in the art that the scope of the disclosure herein is not limited to the particular combination of features described above, but also encompasses other combinations of features described above or their equivalents without departing from the spirit of the disclosure. For example, the above features may be replaced with (but not limited to) features having similar functions disclosed in the present application.
Claims (10)
1. A deep eutectic solvent comprising at least two of DL-menthol, thymol, camphor and alpha-terpineol.
2. The deep eutectic solvent of claim 1, wherein the deep eutectic solvent comprises DL-menthol and thymol, or DL-menthol and camphor, or DL-menthol and alpha-terpineol, or thymol and camphor, or thymol and alpha-terpineol, or camphor and alpha-terpineol.
3. The deep eutectic solvent according to claim 2, characterized in that it comprises DL-menthol and thymol in a molar ratio of 1.
4. The deep eutectic solvent of any one of claims 1 to 3, wherein said deep eutectic solvent is in a liquid state at room temperature.
5. A process for preparing the deep eutectic solvent as claimed in any one of claims 1 to 4, wherein the raw materials are uniformly mixed at 70 to 90 ℃ in predetermined amounts to form the deep eutectic solvent in a liquid state.
6. Use of the deep eutectic solvent of any one of claims 1 to 4 for the extraction of astaxanthin.
7. A method for extracting astaxanthin, which comprises the following steps:
mixing raw materials including astaxanthin source materials and the deep eutectic solvent so as to obtain an astaxanthin extracting solution.
8. The method according to claim 7, wherein the solid-to-liquid ratio of the astaxanthin source material to the deep eutectic solvent is 1.
9. The method according to claim 7, wherein the raw materials including astaxanthin source material and the deep eutectic solvent are mixed and subjected to a shaking process;
optionally, the oscillation speed is 800-1200 rmp, and the time is 0.5-3 h;
optionally, the shaking treatment is performed at a temperature of 40 to 80 ℃.
10. The method according to any one of claims 7 to 8, wherein the source material of astaxanthin comprises Haematococcus pluvialis;
optionally, the haematococcus pluvialis is subjected to cell disruption treatment before being mixed with the deep eutectic solvent;
optionally, hydrochloric acid is adopted for cell disruption treatment, the concentration of the hydrochloric acid is 2-5M, the mass ratio of haematococcus pluvialis to the hydrochloric acid is 1 (5-15), the treatment time is 2min, and the treatment temperature is 50-80 ℃.
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| CN116042309A (en) * | 2023-02-03 | 2023-05-02 | 山东康境海洋生物工程有限公司 | Extraction method of astaxanthin-rich krill oil |
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| US20070196894A1 (en) * | 2006-02-21 | 2007-08-23 | SUNGKYUNKWAN UNIVERSITY, Foundation for Corporate Collaboration | Method for selective separation of free-astaxanthin from green algae haematococcus pluvialis |
| CN111359260A (en) * | 2020-03-30 | 2020-07-03 | 南京林业大学 | Hydrophobic deep eutectic solvent for removing ginkgolic acid in crude ginkgo leaf extract by liquid-liquid micro extraction, and preparation method and removal method thereof |
| CN113912527A (en) * | 2021-10-12 | 2022-01-11 | 自然资源部第三海洋研究所 | Method for extracting astaxanthin by adopting hydrophobic eutectic solvent |
| CN115043766A (en) * | 2022-07-13 | 2022-09-13 | 山西农业大学 | Method for continuously extracting beta-carotene by using eutectic solvent and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| US20070196894A1 (en) * | 2006-02-21 | 2007-08-23 | SUNGKYUNKWAN UNIVERSITY, Foundation for Corporate Collaboration | Method for selective separation of free-astaxanthin from green algae haematococcus pluvialis |
| CN111359260A (en) * | 2020-03-30 | 2020-07-03 | 南京林业大学 | Hydrophobic deep eutectic solvent for removing ginkgolic acid in crude ginkgo leaf extract by liquid-liquid micro extraction, and preparation method and removal method thereof |
| CN113912527A (en) * | 2021-10-12 | 2022-01-11 | 自然资源部第三海洋研究所 | Method for extracting astaxanthin by adopting hydrophobic eutectic solvent |
| CN115043766A (en) * | 2022-07-13 | 2022-09-13 | 山西农业大学 | Method for continuously extracting beta-carotene by using eutectic solvent and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116042309A (en) * | 2023-02-03 | 2023-05-02 | 山东康境海洋生物工程有限公司 | Extraction method of astaxanthin-rich krill oil |
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