Method for extracting isochlorogenic acid C from folium artemisiae argyi
By using a deep eutectic solvent-enzyme composite cell disruption method coupled with magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP, the problems of high energy consumption and poor selectivity in the traditional process of extracting isochlorogenic acid C from Artemisia argyi have been solved. This method achieves efficient and low-cost extraction and purification of isochlorogenic acid C, significantly improving product purity and recovery rate, and meeting green production standards.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-11
- Publication Date
- 2026-03-27
AI Technical Summary
Traditional processes for extracting isochlorogenic acid C from Artemisia argyi are energy-intensive, have poor selectivity, involve highly toxic solvents, are cumbersome, result in low product purity, and cause easy degradation of active ingredients, making it difficult to achieve efficient and low-cost large-scale production.
A method combining deep eutectic solvent-enzyme composite cell disruption with magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP was adopted to achieve efficient extraction and purification of isochlorogenic acid C through synergistic cell disruption, selective adsorption, and low-temperature desorption.
It achieves low-temperature, high-efficiency, and highly selective extraction and purification, with product purity ≥95% and recovery rate ≥92%, reducing energy consumption and solvent consumption, and meeting the requirements of green production.
Abstract
Description
Technical Field
[0001] This invention relates to the technical field of plant extracts, and more particularly to a method for extracting isochlorogenic acid C from Artemisia argyi leaves. Background Technology
[0002] Isochlorogenic acid C is a highly representative polyphenolic acid active ingredient in Artemisia argyi, possessing multiple physiological activities such as antioxidant, anti-inflammatory, hepatoprotective, and glucose and lipid metabolism regulation, and is widely used in functional foods, herbal medicines, and cosmetic formulations. However, the cell walls of Artemisia argyi are composed of a dense cellulose-lignin network, and isochlorogenic acid C exists in both bound and free states within the mesophyll cells. Traditional extraction methods typically employ high-temperature reflux or prolonged extraction with a single organic solvent, which is not only energy-intensive and time-consuming but also highly prone to polyphenol oxidation, isomerization, and degradation, resulting in low recovery rates of the target component and significant batch-to-batch variations. Subsequent purification steps using liquid-liquid extraction, macroporous resins, or column chromatography are cumbersome, consume large amounts of organic solvents, and present significant challenges in solvent residue and recovery. Furthermore, the lack of highly selective separation materials for isochlorogenic acid C makes it difficult to efficiently remove other chlorogenic acid isomers and co-extractants such as flavonoids and polysaccharides, thus limiting product purity. Furthermore, scaling up traditional extraction processes significantly increases equipment investment and operating costs, and also raises the pressure of solvent waste treatment, which is detrimental to green production and energy conservation and emission reduction requirements. Therefore, developing an isochlorogenic acid C extraction and purification technology that is mild, highly selective, requires less solvent, and is easy to scale up has become a key common problem that urgently needs to be solved in the high-value utilization of Artemisia argyi. Summary of the Invention
[0003] In view of the shortcomings of the prior art, the present invention provides a method for extracting isochlorogenic acid C from Artemisia argyi leaves, so as to solve the technical problems of high energy consumption, poor selectivity, high solvent toxicity, cumbersome steps, low product purity and easy degradation of active ingredients in traditional high temperature reflux extraction.
[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows: A method for extracting isochlorogenic acid C from Artemisia argyi leaves, the method comprising the following steps: S1: Grind dried mugwort leaves into powder of 20-80 mesh to obtain mugwort powder; S2: The Artemisia argyi powder and the deep eutectic solvent-enzyme composite cell-wall breaking solution system are mixed at a material-liquid ratio of 1:10 to 1:30 g / mL to perform synergistic cell-wall breaking, and a cell-wall breaking mixture is obtained. S3: Add magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP (IsoC-MIP refers to isochlorogenic acid C molecularly imprinted polymer) to the cell wall-breaking mixture to selectively adsorb isochlorogenic acid C, and then perform magnetic separation with a permanent magnet to obtain magnetic particles loaded with isochlorogenic acid C. S4: Desorb the magnetic particles with an eluent, collect the eluent, concentrate under reduced pressure, and freeze-dry to obtain isochlorogenic acid C product with a purity ≥95%. As a preferred technical solution, the deep eutectic solvent-enzyme composite cell-wall breaking solution system consists of the following components: a) a deep eutectic solvent, which is a low eutectic mixture formed by choline chloride and glycerol in a molar ratio of 1:1.5 to 1:3, with a water content of 15% to 35%; b) cellulase and / or pectinase, with a total enzyme activity of 5 to 20 U / mL; c) 0.05% to 0.2 wt% antioxidant, selected from ascorbic acid or citric acid. As a preferred technical solution, the pH of the deep eutectic solvent-enzyme composite cell-wall breaking solution system is 4.0-5.5, adjusted by an acetate-sodium acetate buffer solution. As a preferred technical solution, the preparation method of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is as follows: Fe3O4@SiO2 magnetic nanoparticles with vinyl groups on the surface are dispersed in acetonitrile, template molecule isochlorogenic acid C, functional monomer, crosslinking agent and initiator are added, nitrogen is purged to remove oxygen and polymerized at 50-70℃ for 6-24h, and then the template molecule is eluted with methanol-acetic acid mixture to obtain the magnetic molecularly imprinted polymer with isochlorogenic acid C specific recognition site. As a preferred technical solution, the Fe3O4@SiO2 magnetic nanoparticles with vinyl content on the surface are obtained by coating SiO2 with Fe3O4 core through sol-gel method and then modifying it with a vinyl-containing silane coupling agent.
[0005] As a preferred technical solution, the functional monomer is selected from at least two of acrylamide, 4-vinylpyridine, and methacrylic acid.
[0006] As a preferred technical solution, the crosslinking agent is a polyethylene glycol dimethacrylate type crosslinking agent having C2-C6 polyoxyethylene segments and terminal dimethacrylate functional groups, selected from at least one of ethylene glycol dimethacrylate, diethylene glycol dimethacrylate, and triethylene glycol dimethacrylate.
[0007] As a preferred technical solution, in step S3, the Fe3O4@IsoC-MIP is added at a rate of 5-20 mg / mL of the cell-breaking mixture, the oscillation speed is 100-200 rpm, the adsorption temperature is 20-35℃, and the adsorption time is 10-30 min.
[0008] As a preferred technical solution, the reduced pressure concentration temperature is ≤45℃, and the freeze-drying conditions are -50℃~-40℃, 10~30Pa.
[0009] As a preferred technical solution, in step S4, the eluent is a methanol-acetic acid mixture of 95:5 to 90:10 v / v, the elution temperature is 20 to 35°C, and the elution time is 2 to 10 min.
[0010] The beneficial effects of this invention are: This invention provides a method for extracting isochlorogenic acid C from Artemisia argyi leaves. It innovatively couples "deep eutectic solvent-enzyme synergistic cell disruption" with "selective adsorption by magnetically imprinted polymers" into a one-step green extraction platform. This overcomes the technical bottlenecks of high energy consumption, solvent toxicity, and poor selectivity associated with the traditional high-temperature reflux-multi-step extraction-column chromatography route, achieving low-temperature, high-efficiency, high-purity, and recyclable continuous extraction. Specifically, Step S1, by pulverizing dried Artemisia argyi leaves into a narrow particle size range of 20-80 mesh, significantly increases the specific surface area and breakage rate of the cell walls, allowing the subsequent solvent-enzyme system to fully penetrate into the cells. On the other hand, it avoids the problems of high viscosity, difficulty in filtration, and excessive oxidation of active ingredients caused by excessively fine powder. This improves the accessibility and extraction efficiency of isochlorogenic acid C at the source and provides a stable physical basis for subsequent homogenization and repeatable cell wall disruption. It solves the technical problem of low mass transfer efficiency and large batch-to-batch differences caused by excessively wide particle size distribution in traditional extraction.
[0011] Step S2 utilizes a choline chloride-glycerol eutectic solvent in synergy with cellulase / pectinase. By leveraging the "swelling-dissolving" effect of the eutectic solvent on the cell wall cellulose-lignin network, which complements the enzyme's directional shearing of glycosidic bonds, rapid cell wall disruption can be achieved under mild conditions. This significantly reduces energy consumption, avoids high-temperature thermosensitive degradation, and maintains enzyme activity through a buffer environment. This allows isochlorogenic acid C to be efficiently released and stably present in the system during the cell wall disruption stage. This overcomes the drawbacks of traditional high-temperature reflux or single organic solvent cell wall disruption, such as high energy consumption, enzyme activity loss, and easy oxidation of polar polyphenols. It achieves a green, efficient, and mild pretreatment effect.
[0012] Step S3 introduces a magnetic molecularly imprinted polymer, Fe3O4@IsoC-MIP, based on surface-vinylened Fe3O4@SiO2. Utilizing the specific cavities on its surface to recognize isochlorogenic acid C, the target molecule can be selectively adsorbed from a complex cell-wall-broken liquid in one step under mild conditions. Magnetic separation rapidly completes solid-liquid separation, avoiding the drawbacks of traditional macroporous resins or liquid-liquid extraction, which require multiple centrifugation, filtration, and large solvent consumption. This significantly improves separation efficiency and product purity. Furthermore, the magnetic particles can be reused more than 20 times, reducing raw material and waste liquid treatment costs and solving common industry problems such as poor selectivity, cumbersome procedures, and high solvent consumption.
[0013] Step S4 employs methanol-acetic acid (95:5~90:10, v / v) for rapid low-temperature desorption, which quantitatively elutes isochlorogenic acid C. Subsequently, it is concentrated under reduced pressure at ≤45℃ and freeze-dried under low-temperature vacuum, avoiding the thermal degradation of phenolic acids and solvent residues caused by high-temperature concentration. The final product has an HPLC purity of ≥95% and a recovery rate of ≥92%. Moreover, no toxic organic extractants are used in the entire process, which meets the quality requirements for green pharmaceuticals and the production of functional food ingredients. This solves the technical bottlenecks of low product purity, high solvent residue, and large activity loss in traditional extraction-purification routes, and achieves a clean production effect with high purity, high recovery, and scalability. Detailed Implementation
[0014] The following description is intended to disclose the invention and enable those skilled in the art to implement it. The preferred embodiments described below are merely examples, and other obvious variations will occur to those skilled in the art.
[0015] Example 1 This embodiment describes a method for extracting isochlorogenic acid C from Artemisia argyi leaves, which includes the following steps: S1: After drying the dried mugwort leaves in a 50℃ forced-air drying oven until constant weight, pulverize them with a high-speed universal pulverizer and pass them through a 60-mesh sieve to obtain mugwort powder with uniform particle size distribution. S2: Weigh 100g of Artemisia argyi powder according to a material-liquid ratio of 1:20g / mL, add 2000mL of a pre-prepared deep eutectic solvent-enzyme complex cell wall breaking solution. This solution is made by mixing choline chloride and glycerol in a molar ratio of 1:2, adding water to adjust the water content to 25%, and then adjusting the pH to 5.0 with acetate-sodium acetate buffer. Then add a complex enzyme of cellulase and pectinase with a total enzyme activity of 10U / mL (mass ratio of 1:1) and 0.1wt% ascorbic acid, and place it in a constant temperature shaker at 35℃ and 150rpm for synergistic cell wall breaking for 30min to obtain a cell wall breaking mixture. S3: The cell wall-breaking mixture was directly transferred into the adsorption container, and 30g of Fe3O4@IsoC-MIP magnetic molecularly imprinted polymer (15mg / mL) was added. After adsorption by shaking at 25℃ and 150rpm for 20min, magnetic separation was completed within 30s using a permanent magnet. The supernatant was discarded to obtain magnetic particles loaded with isochlorogenic acid C. S4: Elute with 200 mL of methanol-acetic acid (95:5, v / v) at 25 °C and 150 rpm for 5 min, shake and desorb, collect the eluent by magnetic separation, repeat the elution once, combine the eluents, concentrate to near dryness under reduced pressure at 40 °C and -0.09 MPa, and then dry in a freeze dryer at -45 °C and 20 Pa for 12 h to obtain pale yellow isochlorogenic acid C lyophilized powder.
[0016] The magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is prepared as follows: Fe3O4@SiO2 magnetic nanoparticles with vinyl groups on their surface are dispersed in acetonitrile, and template molecules isochlorogenic acid C, functional monomers, crosslinking agents and initiators are added. After nitrogen purging to remove oxygen, the polymer is polymerized at 50-70℃ for 6-24 hours. Then, the template molecules are eluted with a methanol-acetic acid mixture to obtain a magnetic molecularly imprinted polymer with specific recognition sites of isochlorogenic acid C. Specifically, firstly, FeCl3·6H2O (0.05 mol) and FeCl3·4H2O (0.025 mol) were dissolved in 200 mL of deionized water at a molar ratio of 2:1. The mixture was heated to 80 °C under nitrogen protection, and 25 mL of 25% NH3·H2O was added at once, instantly forming a black precipitate. The mixture was stirred for another 30 min. After the reaction was completed, the mixture was magnetically separated, washed with deionized water until neutral, and vacuum dried to obtain Fe3O4 nanoparticles with a particle size of 10-20 nm and a saturation magnetization of ≥55 emu / g. Next, the Fe3O4 nanoparticles were washed with deionized water until neutral and then dispersed in a mixture of ethanol / water / ammonia (volume ratio 80:15:5). Tetraethyl orthosilicate (mass ratio of Fe3O4 to Fe3O4 1.2:1) was added dropwise under stirring at room temperature to form a sol-gel coating. After reacting for 12 h, Fe3O4@SiO2 was obtained. Subsequently, the product was redispersed in anhydrous toluene, and γ-methacryloyloxypropyltrimethoxysilane (mass ratio of SiO2 0.3:1) was added under nitrogen protection. The mixture was refluxed at 110 °C for 18 h, washed with ethanol, and vacuum dried to obtain Fe3O4@SiO2 magnetic nanoparticles with vinyl groups on the surface. Finally, 1 g of the above vinylated Fe3O4@SiO2 was weighed and dispersed in 50 mL of acetonitrile. Then, 0.5 mmol of the template molecule isochlorogenic acid C, 1 mmol each of the functional monomers acrylamide and 4-vinylpyridine (molar ratio 1:1), 4 mmol of the crosslinking agent triethylene glycol dimethacrylate, and 20 mg of the initiator AIBN were added sequentially. After ultrasonic degassing for 10 min, nitrogen was purged for 30 min, and the mixture was sealed and placed in a 60 °C water bath for polymerization for 12 h. After the reaction was completed, magnetic separation was performed, and Soxhlet extraction was performed with methanol:acetic acid = 9:1 (v / v) until no template leakage was detected by HPLC. The mixture was then dried under vacuum at 60 °C to obtain brownish-yellow Fe3O4@IsoC-MIP with a particle size of 180-220 nm.
[0017] Test results show that the purity of the final product isochlorogenic acid C reaches 96.2%, the recovery rate is 93.5%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 90% after 20 cycles.
[0018] Example 2 This embodiment describes a method for extracting isochlorogenic acid C from Artemisia argyi leaves, which includes the following steps: S1: After drying the dried mugwort leaves in a vacuum drying oven at 55℃ until constant weight, pulverize them with a high-speed universal pulverizer and pass them through a 40-mesh sieve to obtain mugwort powder with uniform particle size distribution. S2: Weigh 100g of Artemisia argyi powder according to a material-liquid ratio of 1:15g / mL, add 1500mL of a pre-prepared deep eutectic solvent-enzyme complex cell wall breaking solution. This solution is made by mixing choline chloride and glycerol in a molar ratio of 1:2.5, then adding water to adjust the water content to 30%, and then adjusting the pH to 4.5 with acetate-sodium acetate buffer. Subsequently, add a complex enzyme of cellulase and pectinase with a total enzyme activity of 15U / mL (mass ratio of 2:1) and 0.15wt% citric acid, and place it in a constant temperature shaker at 32℃ and 180rpm for 25min to synergistically break the cell wall and obtain the cell wall breaking mixture. S3: The cell wall-breaking mixture was directly transferred into the adsorption container, and 15g of 10mg / mL magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP was added. After adsorption by shaking at 30℃ and 180rpm for 15min, magnetic separation was completed within 30s using a permanent magnet. The supernatant was discarded to obtain magnetic particles loaded with isochlorogenic acid C. S4: Elute with 150 mL of methanol-acetic acid (92:8, v / v) at 30 °C and 180 rpm for 3 min, shake and desorb, collect the eluent by magnetic separation, repeat the elution once, combine the eluents, concentrate to near dryness under reduced pressure at 38 °C and -0.095 MPa, and then dry in a freeze dryer at -42 °C and 15 Pa for 10 h to obtain pale yellow isochlorogenic acid C lyophilized powder.
[0019] The preparation method of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this example, the functional monomer is replaced by 2 mmol of acrylamide and methacrylic acid (molar ratio 1:1), the crosslinking agent is replaced by 3.5 mmol of diethylene glycol dimethacrylate, the polymerization temperature is set to 65℃, and the time is shortened to 10h; the resulting brownish-yellow Fe3O4@IsoC-MIP has a particle size of 190-230nm.
[0020] Test results show that the purity of the final product isochlorogenic acid C reaches 95.8%, the recovery rate is 92.1%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 90% after 20 cycles.
[0021] Example 3 This embodiment describes a method for extracting isochlorogenic acid C from Artemisia argyi leaves, which includes the following steps: S1: After drying the air-dried mugwort leaves in a 45℃ hot air drying oven until constant weight, pulverize them with a high-speed shear pulverizer and pass them through a 50-mesh sieve to obtain mugwort powder with uniform particle size. S2: Weigh 100g of Artemisia argyi powder according to a material-liquid ratio of 1:25g / mL, add 2500mL of a pre-prepared deep eutectic solvent-enzyme complex cell wall breaking solution. This solution is made by mixing choline chloride and glycerol in a molar ratio of 1:1.8, then adding water to adjust the water content to 20%, and then adjusting the pH to 4.2 with citrate-sodium citrate buffer. Subsequently, add a complex enzyme of cellulase and pectinase with a total enzyme activity of 12U / mL (mass ratio 1:1) and 0.08wt% ascorbic acid, and place it in a constant temperature shaker at 38℃ and 120rpm for 35min to synergistically break the cell wall and obtain a cell wall breaking mixture. S3: The cell wall-breaking mixture was directly transferred into the adsorption container, and 12 mg / mL of magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP25 g was added. After adsorption by shaking at 22℃ and 120 rpm for 25 min, magnetic separation was completed within 30 s using a permanent magnet. The supernatant was discarded to obtain magnetic particles loaded with isochlorogenic acid C. S4: Elute with 250 mL of methanol-acetic acid (92:8, v / v) at 22 °C and 120 rpm for 6 min, and collect the eluent by magnetic separation. Repeat the elution once and combine the eluents. Concentrate to near dryness under reduced pressure at 42 °C and -0.085 MPa, and then dry in a freeze dryer at -48 °C and 18 Pa for 14 h to obtain pale yellow isochlorogenic acid C lyophilized powder.
[0022] The preparation method of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this example, the functional monomer is 1.5 mmol of 4-vinylpyridine and methacrylic acid (molar ratio 2:1), the crosslinking agent is 4.5 mmol of ethylene glycol dimethacrylate, the initiator is 25 mg of benzoyl peroxide (BPO), the polymerization temperature is 70℃, and the time is 8 h; the final product particle size is 170-210 nm.
[0023] Test results show that the purity of the final product isochlorogenic acid C reaches 95.6%, the recovery rate is 92.8%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity retention rate of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 89% after 20 cycles.
[0024] Example 4 The method for extracting isochlorogenic acid C from Artemisia argyi in this embodiment is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this embodiment, ethylene glycol dimethacrylate is used instead of triethylene glycol dimethacrylate as a crosslinking agent.
[0025] Test results show that the purity of the final product isochlorogenic acid C reaches 95.3%, the recovery rate is 92.5%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity retention rate of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 90% after 20 cycles.
[0026] Example 5 The method for extracting isochlorogenic acid C from Artemisia argyi in this embodiment is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this embodiment, diethylene glycol dimethacrylate is used instead of triethylene glycol dimethacrylate as a crosslinking agent.
[0027] Test results show that the purity of the final product isochlorogenic acid C reaches 95.5%, the recovery rate is 93.0%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity retention rate of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 91% after 20 cycles.
[0028] Example 6 The method for extracting isochlorogenic acid C from Artemisia argyi in this embodiment is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this embodiment, citric acid is used instead of ascorbic acid as an antioxidant.
[0029] Test results show that the purity of the final product isochlorogenic acid C reaches 95.8%, the recovery rate is 93.2%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity retention rate of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 90% after 20 cycles.
[0030] Example 7 The method for extracting isochlorogenic acid C from Artemisia argyi in this embodiment is basically the same as that in Example 1 in terms of raw material composition and preparation steps. The difference is that in the preparation method of this embodiment, the functional monomer is composed of acrylamide, 4-vinylpyridine and methacrylic acid in a molar ratio of 1:1:1.
[0031] Test results show that the purity of the final product isochlorogenic acid C reaches 97.2%, the recovery rate is 93.5%, the magnetic separation process can be completed within 30 seconds, and the adsorption capacity retention rate of the magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is 92% after 20 cycles.
[0032] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.
Claims
1. A method for extracting isochlorogenic acid C from Artemisia argyi leaves, characterized in that, The method includes the following steps: S1: Grind dried mugwort leaves into powder of 20-80 mesh to obtain mugwort powder; S2: The Artemisia argyi powder and the deep eutectic solvent-enzyme composite cell-wall breaking solution system are mixed at a material-liquid ratio of 1:10 to 1:30 g / mL to perform synergistic cell-wall breaking, and a cell-wall breaking mixture is obtained. S3: Add magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP to the cell wall-breaking mixture to selectively adsorb isochlorogenic acid C, and then perform magnetic separation with a permanent magnet to obtain magnetic particles loaded with isochlorogenic acid C. S4: Desorb the magnetic particles with an eluent, collect the eluent, concentrate under reduced pressure, and freeze-dry to obtain isochlorogenic acid C product with a purity ≥95%.
2. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The deep eutectic solvent-enzyme composite cell-wall breaking solution system consists of the following components: a) a deep eutectic solvent, which is a low eutectic mixture formed by choline chloride and glycerol in a molar ratio of 1:1.5 to 1:3, with a water content of 15 to 35%; b) cellulase and / or pectinase, with a total enzyme activity of 5 to 20 U / mL; c) 0.05 to 0.2 wt% antioxidant, selected from ascorbic acid or citric acid.
3. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The pH of the deep eutectic solvent-enzyme composite cell-breaking solution system is 4.0–5.5, adjusted by an acetate-sodium acetate buffer solution.
4. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The magnetic molecularly imprinted polymer Fe3O4@IsoC-MIP is prepared as follows: Fe3O4@SiO2 magnetic nanoparticles with vinyl groups on their surface are dispersed in acetonitrile, and template molecules isochlorogenic acid C, functional monomers, crosslinking agents and initiators are added. After nitrogen purging to remove oxygen, the polymer is polymerized at 50-70℃ for 6-24 hours. Then, the template molecules are eluted with a methanol-acetic acid mixture to obtain a magnetic molecularly imprinted polymer with specific recognition sites of isochlorogenic acid C.
5. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The vinyl-containing Fe3O4@SiO2 magnetic nanoparticles are obtained by coating SiO2 onto Fe3O4 cores via a sol-gel method, followed by modification with a vinyl-containing silane coupling agent.
6. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The functional monomer is selected from at least two of acrylamide, 4-vinylpyridine, and methacrylic acid.
7. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The crosslinking agent is a polyethylene glycol dimethacrylate type crosslinking agent having C2-C6 polyoxyethylene segments and terminal dimethacrylate functional groups, selected from at least one of ethylene glycol dimethacrylate, diethylene glycol dimethacrylate, and triethylene glycol dimethacrylate.
8. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... In step S3, the Fe3O4@IsoC-MIP is added at a rate of 5–20 mg / mL of the cell-breaking mixture, the shaking speed is 100–200 rpm, the adsorption temperature is 20–35 °C, and the adsorption time is 10–30 min.
9. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... The vacuum concentration temperature is ≤45℃, and the freeze-drying conditions are -50℃~-40℃ and 10~30Pa.
10. The method for extracting isochlorogenic acid C from Artemisia argyi as described in claim 1, characterized in that... In step S4, the eluent is a methanol-acetic acid mixture of 95:5 to 90:10 v / v, the elution temperature is 20 to 35°C, and the elution time is 2 to 10 min.