Separation method of flavonoid compounds in perilla leaves
Through the foam separation method and aminosilane-modified nanofiber separation aid, the problems of low flavonoid extraction rate and high cost in the existing technology are solved, and an efficient and environmentally friendly flavonoid separation effect is achieved.
Patent Information
- Application Number
- CN202510851545.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-24
- Publication Date
- 2025-09-26
AI Technical Summary
The existing technology has problems such as solvent residue in the enriched flavonoid extract, low extraction rate, long production cycle and high cost, making it difficult to achieve an efficient, pollution-free and low-cost separation method.
The foam separation method was adopted, and aminosilane-modified nanofibers were used as separation aids to separate flavonoids from the water extract of Perilla leaves through a foam separation tower.
The method achieves efficient separation of flavonoids from perilla leaves, is simple to operate, low in cost, and environmentally friendly, and improves the extraction rate and purity.
Smart Images
Figure CN120695482A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of foam separation, in particular to a method for separating flavonoid compounds in perilla leaves. Background Art
[0002] Flavonoids are a class of secondary metabolites widely found in the plant kingdom and belong to the polyphenolic class of compounds. They have a unique chemical structure, namely a basic C6-C3-C6 skeleton composed of two benzene rings (Ring A and Ring B) connected by a three-carbon chain (Ring C). This structure gives flavonoids rich chemical properties and diverse biological activities. In plants, flavonoids play a variety of important physiological functions. They can effectively resist the invasion of the external environment. For example, their antioxidant effect can scavenge free radicals produced by metabolic activities in plants and protect cells from oxidative damage. Their anti-ultraviolet function can help plants resist ultraviolet radiation and reduce the damage of ultraviolet rays to plant cells. In addition, flavonoids have anti-pathogenic effects, which can enhance the plant's immunity and resist the invasion of pathogens such as bacteria and fungi.
[0003] Flavonoids not only play a key role in plant physiological processes but also have extensive applications in a wide range of fields, including medicine, food, and cosmetics. In the pharmaceutical field, flavonoids have attracted considerable attention for their diverse biological activities. They possess significant antioxidant capacity, enabling them to scavenge free radicals, slow cellular aging, and prevent the onset of numerous chronic diseases. Their anti-inflammatory properties offer potential applications in the treatment of inflammatory diseases, such as arthritis, by reducing inflammation and alleviating symptoms. Furthermore, flavonoids exhibit anti-tumor activity, inhibiting tumor cell proliferation and inducing apoptosis, offering new insights into cancer prevention and treatment. Their antibacterial and antiviral properties make them of great significance in the development of antimicrobial and antiviral drugs, helping to address the growing challenges of bacterial resistance and viral infections. Their cardiovascular protective effects, including lowering blood lipids, regulating blood pressure, and improving endothelial function, play a positive role in the prevention of cardiovascular disease.
[0004] Among the existing methods for enriching flavonoids in flavonoid extracts, solvent extraction uses solvents to dissolve flavonoids, but this has problems such as solvent residue, low extraction rate, and long production cycles. Precipitation precipitates flavonoids by changing their solubility or adding reagents, but this can lead to loss of active ingredients, making the process time-consuming and costly. Macroporous adsorption resin adsorption of flavonoids by resin has good enrichment effects, but requires high resin selection and regeneration conditions and is relatively expensive. Supercritical fluid extraction has high extraction efficiency and no solvent residue, but the equipment is expensive and difficult to apply on a large scale. Therefore, there is a need to establish a green enrichment method that is efficient, pollution-free, and low-cost.
[0005] In view of this, the present invention is proposed. Summary of the Invention
[0006] The object of the present invention is to provide a method for separating flavonoids from perilla leaves. The method is simple to operate and can efficiently separate flavonoids from perilla leaves to solve the above technical problems.
[0007] In order to achieve the above objectives, the following technical solutions are adopted:
[0008] The present invention provides a method for separating flavonoid compounds from perilla leaves, which uses a foam separation method to separate the flavonoid compounds from the perilla leaf water extract;
[0009] The separation aid for foam separation is aminosilane-modified nanofiber.
[0010] As a further technical solution, the preparation method of the perilla leaf water extract comprises the following steps:
[0011] The perilla leaves are crushed and mixed with water for water extraction. After the water extraction is completed, solid-liquid separation is performed, and the clear liquid is collected to obtain the perilla leaf water extract.
[0012] As a further technical solution, the water extraction is carried out under ultrasonic conditions;
[0013] The material-liquid ratio of the water extraction is 1g:10mL-1g:50mL, the temperature is 20-45°C, the ultrasonic power is 60-300W, and the extraction time is 50-100min.
[0014] As a further technical solution, the preparation method of the aminosilane-modified nanofibers is as follows:
[0015] An aminosilane reagent and nanofibers are mixed in a solution, and aminosilane-modified nanofibers are prepared after a modification reaction.
[0016] As a further technical solution, the aminosilane reagent includes 3-aminopropyltriethoxysilane (APTES).
[0017] As a further technical solution, the nanofibers are extracted from perilla straw.
[0018] As a further technical solution, the following steps are included:
[0019] The water extract of perilla leaves and aminosilane-modified nanofibers are mixed to obtain a feed liquid, which is then injected into a foam separation tower for foam separation. When the foam can no longer flow out of the top of the foam separation tower, ventilation is stopped to complete the separation of flavonoid compounds in the perilla leaves.
[0020] As a further technical solution, the concentration of the aminosilane-modified nanofibers in the feed solution is 100-600 mg / L.
[0021] As a further technical solution, the pH of the foam separation is 3-5.
[0022] As a further technical solution, the gas velocity of the foam separation is 300 to 500 mL / min.
[0023] Compared with the prior art, the present invention has the following beneficial effects:
[0024] The present invention provides a method for separating flavonoids from perilla leaves. The method uses aminosilane-modified nanofibers as a separation aid. The nanofibers specifically adsorb flavonoids and have excellent foaming properties, enabling separation of the flavonoids through foam separation. The method is simple to operate, low-cost, and environmentally friendly, enabling efficient separation of flavonoids from perilla leaves. BRIEF DESCRIPTION OF THE DRAWINGS
[0025] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.
[0026] Figure 1 The effect of solid-liquid ratio on the extraction of flavonoids from Perilla leaves;
[0027] Figure 2 The effect of temperature on the extraction of flavonoids from Perilla leaves;
[0028] Figure 3 The effect of extraction time on the extraction of flavonoids from Perilla leaves;
[0029] Figure 4 The effect of ultrasonic power on the extraction of flavonoids from Perilla leaves;
[0030] Figure 5 Response surface study for the preparation of perilla leaf extract;
[0031] Figure 6 This is an electron micrograph of aminosilane-modified nanofibers;
[0032] Figure 7 The effect of the concentration of aminosilane-modified nanofibers on the foam separation of flavonoids from Perilla leaves;
[0033] Figure 8The effect of pH on the foam separation of flavonoids from Perilla leaves;
[0034] Figure 9 The effect of gas velocity on foam separation of flavonoids from Perilla leaves;
[0035] Figure 10 This study is about the effect of extraction volume on foam separation of flavonoids from Perilla leaves. DETAILED DESCRIPTION
[0036] Below in conjunction with embodiment and example, embodiment of the present invention is described in detail, but those skilled in the art will appreciate that the following embodiment and example are only used to illustrate the present invention, and should not be considered as limiting the scope of the present invention. Based on the embodiment in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work premise all fall within the scope of protection of the present invention. Unspecified conditions are carried out according to the conditions of normal conditions or manufacturer's recommendations. Reagents used or instruments not specified by the manufacturer are conventional products that can be purchased commercially.
[0037] The present invention provides a method for separating flavonoid compounds from perilla leaves, which uses a foam separation method to separate the flavonoid compounds from the perilla leaf water extract;
[0038] The separation aid for foam separation is aminosilane-modified nanofiber.
[0039] The inventors have discovered that aminosilane-modified nanofibers can effectively adsorb and capture flavonoids and have excellent foaming properties, enabling the separation of flavonoids through foam separation. The separation method provided by the present invention, using aminosilane-modified nanofibers as a separation aid, is simple to operate, low-cost, and environmentally friendly, enabling the efficient separation of flavonoids from perilla leaves.
[0040] In some optional embodiments, the method for preparing the perilla leaf water extract comprises the following steps:
[0041] The perilla leaves are crushed and mixed with water for water extraction. After the water extraction is completed, solid-liquid separation is performed, and the clear liquid is collected to obtain the perilla leaf water extract.
[0042] In some optional embodiments, the water extraction is performed under ultrasonic conditions;
[0043] The material-liquid ratio of the water extraction can be, for example, but not limited to 1g:10mL, 1g:30mL or 1g:50mL, preferably 1g:30mL; the temperature can be, for example, but not limited to 20°C, 30°C or 45°C, preferably 35°C; the ultrasonic power can be, for example, but not limited to 60W, 150W or 300W, preferably 300W; the extraction time can be, for example, but not limited to 50min, 75min or 100min, preferably 80min.
[0044] Since aminosilane modification can not only change the hydrophilic properties of nanocellulose, but also enhance the selective adsorption capacity of nanocellulose to flavonoids (such as through the interaction between the amino functional group and the flavonoid hydroxyl group), the present invention uses a method for modifying nanocellulose with aminosilane. In some optional embodiments, the preparation method of the aminosilane-modified nanofiber is as follows:
[0045] An aminosilane reagent and nanofibers are mixed in a solution, and aminosilane-modified nanofibers are prepared after a modification reaction.
[0046] In some optional embodiments, the aminosilane reagent includes but is not limited to 3-aminopropyltriethoxysilane, or other aminosilane reagents well known to those skilled in the art.
[0047] In some optional embodiments, during the modification reaction, the final concentration of the aminosilane reagent is 4 wt%-20 wt%, and the final concentration of the nanocellulose is 0.01 g / mL.
[0048] In some optional embodiments, the nanofibers are extracted from Perilla frutescens straw.
[0049] Nanofibers are renewable, biocompatible, and mechanically strong. They are sourced from perilla straw, have low costs, and improve the utilization rate of perilla.
[0050] In some optional embodiments, the following steps are included:
[0051] The water extract of perilla leaves and aminosilane-modified nanofibers are mixed to obtain a feed liquid, which is then injected into a foam separation tower for foam separation. When the foam can no longer flow out of the top of the foam separation tower, ventilation is stopped to complete the separation of flavonoid compounds in the perilla leaves.
[0052] In some optional embodiments, the concentration of the aminosilane-modified nanofibers in the feed solution may be, for example, but not limited to, 100 mg / L, 300 mg / L, or 600 mg / L.
[0053] In some optional embodiments, the pH of the foam separation can be, for example, but not limited to, 3, 4, or 5.
[0054] In some optional embodiments, the gas velocity for foam separation may be, for example, but not limited to, 300 mL / min, 400 mL / min, or 500 mL / min.
[0055] The present invention is further described below by means of specific examples and comparative examples. However, it should be understood that these examples are merely provided for more detailed description and are not to be construed as limiting the present invention in any form.
[0056] Example 1 Effect of material-liquid ratio on the extraction of flavonoids from Perilla leaves
[0057] Perilla leaves were crushed to prepare perilla powder, which was then mixed with water at a ratio of 1:10-1:50 (g / mL) in a constant temperature water bath at 30°C. The mixture was extracted at an ultrasonic power of 180 W for 30 min, and the waste residue was removed by centrifugation at 10,000 rpm for 15 min. The perilla flavonoid extract was then filtered under reduced pressure, and the flavonoid concentration was detected by spectrophotometry.
[0058] The results are as follows Figure 1 As shown in the figure, when the solid-liquid ratio was 1:(20-40) (g / mL), the extraction rate of flavonoids was better.
[0059] Therefore, the solid-liquid ratio of the flavonoid extract used in the subsequent examples was 1:30 (g / mL).
[0060] Example 2 Effect of Temperature on the Extraction of Flavonoids from Perilla Leaves
[0061] Perilla leaves were crushed to prepare perilla powder, which was then mixed with water at a ratio of 1:30 (g / mL) and placed in a constant temperature water bath at 15-55°C. The mixture was extracted at an ultrasonic power of 180 W for 30 min, and the waste residue was removed by centrifugation at 10,000 rpm for 15 min. The perilla flavonoid extract was then filtered under reduced pressure and the flavonoid concentration was determined by spectrophotometry.
[0062] The results are as follows Figure 2 As shown in the figure, when the temperature is 35℃, the extraction rate of flavonoids is better.
[0063] Therefore, the flavonoid extracts used in the subsequent examples were all extracted at 35°C.
[0064] Example 3 Effect of extraction time on the extraction of flavonoids from Perilla leaves
[0065] Perilla leaves were crushed to prepare perilla powder, which was then mixed with water at a ratio of 1:30 (g / mL) and placed in a 35°C constant temperature water bath. The mixture was extracted at an ultrasonic power of 180 W for 50-100 min, and the waste residue was removed by centrifugation at 10,000 rpm for 15 min. The perilla flavonoid extract was then filtered under reduced pressure and the flavonoid concentration was determined by spectrophotometry.
[0066] The results are as follows Figure 3 As shown in the figure, when the extraction time is 80 min, the extraction rate of flavonoids is better.
[0067] Therefore, the extraction time of the flavonoid extracts used in the subsequent examples was 80 min.
[0068] Example 4 Effect of Ultrasonic Power on the Extraction of Flavonoids from Perilla Leaves
[0069] Perilla leaves were crushed to prepare perilla powder, which was then mixed with water at a ratio of 1:30 (g / mL) and placed in a 35°C constant temperature water bath. The mixture was extracted for 80 min at an ultrasonic power of 60-300 W. The waste residue was removed by centrifugation at 10,000 rpm for 15 min, and the perilla flavonoid extract was obtained by filtration under reduced pressure. The flavonoid concentration was detected by spectrophotometry.
[0070] The results are as follows Figure 4 As shown in the figure, when the power is 300W, the extraction rate of flavonoids is better.
[0071] Therefore, the extraction power of the flavonoid extracts used in the subsequent examples was 300W.
[0072] In summary, the optimal extraction parameters of flavonoid extract are as follows:
[0073] Example 5 Response surface study of flavonoid extract preparation
[0074] The perilla leaves were crushed to prepare perilla powder, which was then mixed with water at a ratio of 1:20-1:40 (g / mL) and placed in a constant temperature water bath at 25-55°C. The mixture was extracted at an ultrasonic power of 180-300W for 70-90 minutes. The waste residue was removed by centrifugation at 10,000 rpm for 15 minutes, and the perilla flavonoid extract was obtained by vacuum filtration. The flavonoid concentration was detected by spectrophotometry.
[0075] The response surface results are as follows Figure 5 As shown, it can be seen that the optimal extraction conditions are as follows:
[0076] The solid-liquid ratio was 1:30 (g / mL), the extraction temperature was 35°C, the extraction time was 80 min, and the extraction power was 300W.
[0077] Therefore, the flavonoid extract used in the subsequent examples will be prepared by the following method:
[0078] Perilla leaves were crushed to produce perilla powder, which was then mixed with water at a ratio of 1:30 (g / mL) in a 35°C constant temperature water bath. Ultrasonic extraction was performed at 300W for 80 minutes, followed by centrifugation at 10,000 rpm for 15 minutes to remove waste residue. The extract was then filtered under reduced pressure to obtain a flavonoid extract, which was then assayed spectrophotometrically for flavonoid concentration. The extraction yield of the flavonoids in the flavonoid extract was 1.808%, with a purity of 10.38%, determined by dividing the mass of the extracted flavonoids by the mass of the perilla leaves.
[0079] Example 6 Preparation of Perilla Straw Cellulose CNF
[0080] Perilla frutescens straw planted in the experimental field of Jinzhong Industrial Technology Research Institute of North University of Shanxi Province was taken, dried and crushed, and passed through an 80-mesh sieve. The powder was then mixed with 4% NaOH solution in a ratio of 1:40 (g / mL), heated in a water bath at 80°C for 4 h, washed with deionized water until the supernatant was clear, and dried for later use.
[0081] Choline chloride (CHCL) and oxalic acid dihydrate (AD) were stirred at 80 ° C for 2 hours in a molar ratio of 1:1 to make it into a transparent liquid, namely DES. The powder dried in the previous step and DES were mixed in a ratio of 1:20 (g / mL), in an oil bath at 120 ° C for 4 hours, cooled and washed with deionized water, centrifuged at 4000 rpm for 10 minutes, homogenized at 10000 rpm for 5 minutes, ultrasonicated at 25 ° C and 300 W for 2 hours, then freeze-dried and ground for use to obtain CNF. The electron microscope image is as follows Figure 6 shown.
[0082] Example 7 Preparation of Aminosilanized Nanocellulose (Si-CNF)
[0083] The CNF prepared in Example 6 was modified with aminosilane. The selected aminosilane reagent was 3-aminopropyltriethoxysilane (APTES), which was diluted to 4%, 8%, 12%, 16%, and 20% aminosilane reagent according to mass concentration, and its pH was adjusted to 5. 0.1 g of nanocellulose was added to 10 mL of the above aminosilane reagent, placed on a magnetic stirring device, stirred at 800 rpm and 25°C for 2 hours, and then centrifuged at 7000 rpm for 12 minutes. The mixture was rinsed with ethanol, dried, and ground into powder for later use. The same method is applicable to the remaining aminosilane reagents.
[0084] After testing the contact angle and foam performance of five kinds of particles, it was found that the nanocellulose modified with 16% aminosilane had the best performance. Therefore, the nanocellulose modified with 16% aminosilane was selected for foam separation of flavonoids.
[0085] Example 8 Effect of the Material-Liquid Ratio of 16%-Si-CNF to Flavonoid Extract on the Foam Separation of Flavonoids
[0086] Take the CNF prepared in Example 6, dilute it with APTES to 16%, adjust its pH to 5, take 0.1 g of nanocellulose and add it to 10 mL of the above-mentioned aminosilane reagent. Place it on a magnetic stirring device, stir at 800 rpm and 25°C for 2 h, then centrifuge at 7000 rpm for 12 min, rinse with ethanol, dry, and grind. The obtained particles are recorded as 16%-Si-CNF.
[0087] 0.02, 0.04, 0.06, 0.08, 0.1, and 0.12 g of 16%-Si-CNF were weighed and dissolved in 200 mL of a diluted flavonoid extract containing 400 mg / L flavonoids. The pH of the solution was adjusted to 4. The solution was then transferred to a flotation tower made of transparent organic glass with an inner diameter of 40.0 mm and a height of 700 mm. An electromagnetic air pump was used to bubble air into the tower through a distributor at a rate of 400 mL / min. Bubbles were collected from the top of the flotation tower within 3 minutes. After 3 minutes, when no more bubbles could be released from the top of the tower, a sample of the residual liquid was collected. The crystal violet removal rate (R) and enrichment ratio (E) were calculated using the following formulas:
[0088]
[0089] In formula (1, 2), C0, C f , C r (mg / L) are the concentrations of rhodamine B in the injection solution, foam solution, and residual solution, respectively; Q0, Q f , Q r (mL / min) are the volume flow rates of feed liquid, foam liquid and residual liquid respectively.
[0090] The results are as follows Figure 7 As shown, when the particle concentration is 500 mg / L, both R and E are higher.
[0091] Example 9 Effect of different pH on the separation of flavonoids by 16%-Si-CNF foam
[0092] Weigh 0.1g of 16%-Si-CNF and dissolve it in 200mL of a diluted flavonoid extract with a flavonoid concentration of 400mg / L. Adjust the solution pH to 3, 3.5, 4, 4.5, and 5, respectively. Transfer the solution to a flotation tower and use an electromagnetic air pump to bubble into the tower through a distributor. Adjust the air speed to 400mL / min. Collect the bubbles that bulge out from the top of the flotation tower within 3 minutes. After 3 minutes, when the foam can no longer bulge out from the top of the tower, take a sample of the residual liquid. Calculate the removal rate R and enrichment ratio E of crystal violet. The results are as follows: Figure 8 As shown, when the solution pH is 4, both R and E are higher.
[0093] Example 10 Effect of different gas velocities on the separation of flavonoids by 16%-Si-CNF foam
[0094] Weigh 0.1g of 16%-Si-CNF and dissolve it in 200mL of a diluted flavonoid extract containing 400mg / L flavonoids. Adjust the solution pH to 4 and transfer the solution to a flotation tower. Use an electromagnetic air pump to bubble air into the tower through a distributor. Adjust the air speed to 300, 350, 400, 450, and 500mL / min, respectively. Collect bubbles from the top of the flotation tower within 3 minutes. After 3 minutes, when the foam can no longer be released from the top of the tower, take a sample of the residual liquid. The removal rate R and enrichment ratio E of crystal violet are calculated. The results are as follows: Figure 9 As shown in Figure 3, when the gas rate is 400 mL / min, both R and E are higher.
[0095] Example 11 Effect of different extraction liquid volumes on flavonoid separation by 16%-Si-CNF foam
[0096] Prepare 100mL, 150mL, and 200mL of a mixed solution of 16%-Si-CNF and flavonoid extract, respectively, to a particle concentration of 500mg / L. Dilute the flavonoid concentration in the flavonoid extract to 400mg / L, adjust the solution pH to 4, transfer the solution to a flotation tower, and use an electromagnetic air pump to bubble into the tower through a distributor. Adjust the air speed to 400mL / min, and collect bubbles from the top of the flotation tower within 3 minutes. After 3 minutes, when the foam can no longer be blown out from the top of the tower, take a sample of the residual liquid. The removal rate R and enrichment ratio E of crystal violet are calculated. The results are as follows: Figure 10 As shown, when the volume of the extract was 100 mL, both R and E were higher.
[0097] After separation, the purity of flavonoids reached 26%, a significant increase of 15.62% compared to the previous purity.
[0098] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the above embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.
Claims
1. A method for separating flavonoids from Perilla leaves, characterized in that: The flavonoids in the aqueous extract of Perilla frutescens leaves were separated by foam separation. The separation aid for foam separation is aminosilane-modified nanofiber.
2. The separation method according to claim 1, wherein The preparation method of the perilla leaf water extract comprises the following steps: The perilla leaves are crushed and mixed with water for water extraction. After the water extraction is completed, solid-liquid separation is performed, and the clear liquid is collected to obtain the perilla leaf water extract.
3. The separation method according to claim 2, characterized in that The water extraction is carried out under ultrasonic conditions; The material-liquid ratio of the water extraction is 1g:10mL-1g:50mL, the temperature is 20-45°C, the ultrasonic power is 60-300W, and the extraction time is 50-100min.
4. The separation method according to claim 1, wherein The preparation method of the aminosilane-modified nanofiber is as follows: An aminosilane reagent and nanofibers are mixed in a solution, and aminosilane-modified nanofibers are prepared after a modification reaction.
5. The separation method according to claim 4, characterized in that The aminosilane reagent includes 3-aminopropyltriethoxysilane.
6. The separation method according to claim 4, characterized in that The nanofiber is extracted from perilla straw.
7. The separation method according to claim 1, characterized in that The following steps are involved: The water extract of perilla leaves and aminosilane-modified nanofibers are mixed to obtain a feed liquid, which is then injected into a foam separation tower for foam separation. When the foam can no longer flow out of the top of the foam separation tower, ventilation is stopped to complete the separation of flavonoid compounds in the perilla leaves.
8. The separation method according to claim 7, characterized in that In the feed solution, the concentration of the aminosilane-modified nanofibers is 100-600 mg / L.
9. The separation method according to claim 7, characterized in that The pH of the foam separation is 3-5.
10. The separation method according to claim 7, characterized in that The gas velocity of the foam separation is 300 to 500 mL / min.