Cyclocarya paliurus extraction method
By combining enzymatic hydrolysis with an alternating magnetic field and a eutectic solvent, the problems of low extraction efficiency and poor component selectivity of Cyclocarya paliurus have been solved, achieving efficient and safe extraction of Cyclocarya paliurus components and promoting its application in the modernization and industrialization of traditional Chinese medicine.
Patent Information
- Application Number
- CN202511209129.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-08-27
- Publication Date
- 2025-12-16
AI Technical Summary
Existing extraction processes for Cyclocarya paliurus have problems such as low extraction efficiency, poor component selectivity, high energy consumption, poor safety, and unclear synergistic relationships of functional components, which limit its application in the modernization and industrialization of traditional Chinese medicine.
An extraction method combining enzymatic hydrolysis with alternating magnetic field and eutectic solvent was adopted. The leaves of *Cyclocarya paliurus* were enzymatically hydrolyzed with citrate-phosphate buffer, and the alternating magnetic field was used to enhance the penetration and complexing ability of the eutectic solvent. The extraction process was optimized by combining macroporous resin separation and ethanol elution.
It significantly improved the extraction rate of effective components from Eucommia ulmoides, enhanced extraction selectivity, reduced energy consumption, improved safety, and promoted the synergistic effect of functional components.
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Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the technical field of extraction, in particular to an extraction method of Cyclocarya paliurus. BACKGROUND
[0002] Cyclocarya paliurus, also known as "sweet tea tree", is a deciduous tree of the family Juglandaceae Cyclocarya, mainly distributed in the middle and lower reaches of the Yangtze River in China. Because its leaves are rich in various bioactive components, it is widely used for the auxiliary treatment of diseases such as diabetes, metabolic syndrome, and lipid metabolism disorders. Modern pharmacological studies have shown that Cyclocarya paliurus leaves are rich in functional components such as flavonoids, triterpenes, organic acids, and polysaccharides, and have significant antioxidant, hypoglycemic, and anti-inflammatory biological activities, and are widely used for the development of functional foods and traditional Chinese medicine preparations.
[0003] However, the extraction efficiency of active ingredients of Cyclocarya paliurus is low, the components are severely damaged, and there are many impurities, which has long limited its popularization and application in the modernization and industrialization of traditional Chinese medicine. Traditional extraction processes mainly include decoction, alcohol extraction, ultrasonic-assisted extraction, etc. These methods generally have the following shortcomings:
[0004] (1) The extraction process conditions are harsh, requiring long-time heating or using a large amount of organic solvent, which not only consumes a lot of energy, but also easily causes degradation of heat-sensitive components;
[0005] (2) The selectivity of target components is poor, often accompanied by a large amount of impurities co-extracted, increasing the difficulty of subsequent separation and purification;
[0006] (3) The green safety is poor, and the use of a large amount of alcohol or other organic solvents has safety and environmental hazards;
[0007] (4) The synergistic relationship between functional components is not clear, and most methods cannot specifically improve the extraction efficiency of key functional components such as protocatechuic acid, isoquercitrin, and quercitrin.
[0008] In recent years, with the development of green extraction technology and biocatalysis technology, composite enzymatic hydrolysis has been gradually applied to the release process of natural medicinal plant components due to its high efficiency, mildness, and strong selectivity. By breaking the plant cell wall structure to release the embedded active ingredients, the extraction rate is improved. However, the composition of the enzyme hydrolysis system and the enzyme hydrolysis conditions still need to be further optimized, and the problem of many impurities in the enzyme hydrolysis solution needs to be solved.
[0009] Therefore, it is urgent to develop a green and efficient extraction process system that is easy to standardize. SUMMARY
[0010] Based on the technical problems existing in the background technology, the present application provides an extraction method of Cyclocarya paliurus, which significantly improves the extraction rate of active ingredients in Cyclocarya paliurus.
[0011] The application provides an extraction method of Cyclobalanopsis glaucoides.
[0012] S1: washing, drying and crushing Cyclobalanopsis glaucoides leaves;
[0013] S2: dispersing the crushed Cyclobalanopsis glaucoides leaves in a citric acid-phosphate buffer, and adding a compound enzyme for enzymolysis;
[0014] S3: adding a deep eutectic solvent to supernatant after the enzymolysis, and performing extraction under the action of an alternating magnetic field;
[0015] S4: after the extraction, the solution is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, so that Cyclobalanopsis glaucoides extract is obtained.
[0016] Preferably, the mass ratio of the Cyclobalanopsis glaucoides leaves, the compound enzyme and the citric acid-phosphate buffer is 1:0.01-0.05:10-20.
[0017] Preferably, the compound enzyme is composed of cellulase and pectinase at a mass ratio of 2:1-4.
[0018] Preferably, the enzymolysis temperature is 40-60 DEG C, and the enzymolysis time is 60-120 min.
[0019] Preferably, the deep eutectic solvent is composed of choline chloride, tartaric acid and erythritol at a molar ratio of 1:0.4-0.6:0.4-0.6; and the volume ratio of the supernatant to the deep eutectic solvent is 1:0.1-1.
[0020] Preferably, the alternating magnetic field has a frequency of 40-60 Hz and an intensity of 0.2-0.4 T.
[0021] Preferably, the extraction temperature is 40-50 DEG C, and the extraction time is 30-60 min.
[0022] The application has the beneficial technical effects that:
[0023] The application extracts Cyclocarya paliurus by the way of alternating magnetic field and low eutectic solvent, which significantly improves the extraction rate of effective components in Cyclocarya paliurus. This is because the alternating magnetic field can make tartaric acid, erythritol and choline chloride in the eutectic solvent produce alternating orientation, and enhance the polarizability. The high-frequency molecular shock can break part of the weak hydrogen bonds, and promote the formation of more stable and denser hydrogen bond network in the new conformation. This phenomenon dynamically optimizes the synergistic structure between tartaric acid and erythritol (i.e. the complex hydrogen bond entanglement formed by hydroxyl and carboxyl groups), thereby forming more "temporary associates" and enhancing the complexing and wrapping ability of polyphenols and flavonoid molecules in Cyclocarya paliurus. In addition, the alternating magnetic field introduces micro-disturbance and micro-turbulence in the liquid, which can induce the solvent to have enhanced flowability, reduced viscosity and improved diffusion coefficient in the system involving erythritol and tartaric acid, thereby promoting the solvent to penetrate the plant cell wall more deeply and improving the cleaning and infiltration ability. The dynamic disturbance further amplifies the penetration and solubilization of the eutectic solvent to the cell structure. Finally, the alternating magnetic field can induce the intracellular components that are not completely released by enzymatic hydrolysis to be further released under the dynamic action of the solvent. At this time, the weak acid property of tartaric acid helps to maintain the extracellular-intracellular pH gradient and accelerate the diffusion of components outward, while the multi-hydroxyl sites of erythritol can dynamically bind free components to form complex states to prevent them from being oxidized and inactivated. DETAILED DESCRIPTION
[0024] The application will be further described below in combination with specific examples.
[0025] The cellulase of the application has the model number FDG-2225, and the pectinase has the model number FFG-0658, both of which are purchased from Ningxia Xiasen Industrial Group Co., Ltd.
[0026] Example 1
[0027] The Cyclocarya paliurus leaves are subjected to cleaning, drying and crushing treatment, and then crushed to pass through a 60-mesh sieve. Then the crushed Cyclocarya paliurus leaves are dispersed in a citric acid-phosphate buffer solution, the pH of the enzyme hydrolysis system is controlled to be 5.5, and then a complex enzyme is added for enzyme hydrolysis. The supernatant after enzyme hydrolysis is added with a low eutectic solvent, and extraction is carried out under the action of an alternating magnetic field. After the extraction solution is separated by a macroporous resin, eluted with ethanol and concentrated by rotary evaporation, a Cyclocarya paliurus extract is obtained.
[0028] The mass ratio of the Cyclocarya paliurus leaves, the complex enzyme and the citric acid-phosphate buffer solution is 1:0.03:15.
[0029] The complex enzyme is composed of cellulase and pectinase in a mass ratio of 1:1; the temperature for enzyme hydrolysis is 50℃, and the enzyme hydrolysis time is 90 min.
[0030] The eutectic solvent is composed of choline chloride, tartaric acid and erythritol in a molar ratio of 1:0.5:0.5; the volume ratio of the supernatant to the eutectic solvent is 1:0.5; the frequency of the alternating magnetic field is 50 Hz, and the intensity is 0.3 T; the extraction temperature is 45 ℃, and the extraction time is 45 min.
[0031] Example 2
[0032] The Cyclocarya paliurus leaves are subjected to cleaning, drying and crushing treatment, and then crushed through a 60-mesh sieve; then the crushed Cyclocarya paliurus leaves are dispersed in a citric acid-phosphate buffer solution, the pH of the enzymatic system is controlled to be 5.5, and then a composite enzyme is added for enzymolysis; the eutectic solvent is added to the supernatant after enzymolysis, and extraction is carried out under the action of an alternating magnetic field; after extraction, the solution is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, and then a Cyclocarya paliurus extract is obtained.
[0033] The mass ratio of the Cyclocarya paliurus leaves, the composite enzyme and the citric acid-phosphate buffer solution is 1:0.01:10.
[0034] The composite enzyme is composed of cellulase and pectinase in a mass ratio of 2:1; the enzymolysis temperature is 40 ℃, and the enzymolysis time is 120 min.
[0035] The eutectic solvent is composed of choline chloride, tartaric acid and erythritol in a molar ratio of 1:0.4:0.4; the volume ratio of the supernatant to the eutectic solvent is 1:0.1; the frequency of the alternating magnetic field is 40 Hz, and the intensity is 0.2 T; the extraction temperature is 40 ℃, and the extraction time is 60 min.
[0036] Example 3
[0037] The Cyclocarya paliurus leaves are subjected to cleaning, drying and crushing treatment, and then crushed through a 60-mesh sieve; then the crushed Cyclocarya paliurus leaves are dispersed in a citric acid-phosphate buffer solution, the pH of the enzymatic system is controlled to be 5.5, and then a composite enzyme is added for enzymolysis; the eutectic solvent is added to the supernatant after enzymolysis, and extraction is carried out under the action of an alternating magnetic field; after extraction, the solution is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, and then a Cyclocarya paliurus extract is obtained.
[0038] The mass ratio of the Cyclocarya paliurus leaves, the composite enzyme and the citric acid-phosphate buffer solution is 1:0.05:20.
[0039] The composite enzyme is composed of cellulase and pectinase in a mass ratio of 1:2; the enzymolysis temperature is 60 ℃, and the enzymolysis time is 60 min.
[0040] The eutectic solvent is composed of choline chloride, tartaric acid and erythritol in a molar ratio of 1:0.6:0.6; the volume ratio of the supernatant to the eutectic solvent is 1:1; the frequency of the alternating magnetic field is 60 Hz, and the intensity is 0.4 T; the extraction temperature is 50 ℃, and the extraction time is 30 min.
[0041] Comparative Example 1
[0042] The Cyclocarya paliurus leaves were washed, dried and crushed, and then sieved through a 60-mesh sieve. The crushed Cyclocarya paliurus leaves were then dispersed in a citric acid-phosphate buffer, the pH of the enzymatic system was controlled at 5.5, and a compound enzyme was added for enzymatic hydrolysis. A deep eutectic solvent was added to the supernatant after enzymatic hydrolysis, and extraction was performed under the action of an alternating magnetic field. After the extracted solution was separated by a macroporous resin, eluted with ethanol and concentrated by rotary evaporation, a Cyclocarya paliurus extract was obtained.
[0043] The mass ratio of the Cyclocarya paliurus leaves, the compound enzyme and the citric acid-phosphate buffer was 1:0.03:15.
[0044] The compound enzyme was composed of cellulase and pectinase at a mass ratio of 1:1. The enzymatic hydrolysis temperature was 50°C, and the enzymatic hydrolysis time was 90 min.
[0045] The deep eutectic solvent was composed of choline chloride and erythritol at a molar ratio of 1:1. The volume ratio of the supernatant to the deep eutectic solvent was 1:0.5. The frequency of the alternating magnetic field was 50 Hz, and the intensity was 0.3 T. The extraction temperature was 45°C, and the extraction time was 45 min.
[0046] Comparative Example 2
[0047] The Cyclocarya paliurus leaves were washed, dried and crushed, and then sieved through a 60-mesh sieve. The crushed Cyclocarya paliurus leaves were then dispersed in a citric acid-phosphate buffer, the pH of the enzymatic system was controlled at 5.5, and a compound enzyme was added for enzymatic hydrolysis. A deep eutectic solvent was added to the supernatant after enzymatic hydrolysis, and extraction was performed under the action of an alternating magnetic field. After the extracted solution was separated by a macroporous resin, eluted with ethanol and concentrated by rotary evaporation, a Cyclocarya paliurus extract was obtained.
[0048] The mass ratio of the Cyclocarya paliurus leaves, the compound enzyme and the citric acid-phosphate buffer was 1:0.03:15.
[0049] The compound enzyme was composed of cellulase and pectinase at a mass ratio of 1:1. The enzymatic hydrolysis temperature was 50°C, and the enzymatic hydrolysis time was 90 min.
[0050] The deep eutectic solvent was composed of choline chloride and tartaric acid at a molar ratio of 1:1. The volume ratio of the supernatant to the deep eutectic solvent was 1:0.5. The frequency of the alternating magnetic field was 50 Hz, and the intensity was 0.3 T. The extraction temperature was 45°C, and the extraction time was 45 min.
[0051] Comparative Example 3
[0052] The Cyclobalanopsis gaber leaves are washed, dried and crushed, and then sieved through a 60-mesh sieve; the crushed Cyclobalanopsis gaber leaves are then dispersed in a citric acid-phosphate buffer, the pH of the enzymatic system is controlled at 5.5, and then a compound enzyme is added for enzymolysis; a eutectic solvent is added to the supernatant after the enzymolysis; and the solution after the extraction is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, to obtain the Cyclobalanopsis gaber extract.
[0053] The mass ratio of the Cyclobalanopsis gaber leaves, the compound enzyme and the citric acid-phosphate buffer is 1:0.03:15.
[0054] The compound enzyme is composed of cellulase and pectinase at a mass ratio of 1:1; the enzymolysis temperature is 50℃, and the enzymolysis time is 90 min.
[0055] The eutectic solvent is composed of choline chloride, tartaric acid and erythritol at a molar ratio of 1:0.5:0.5; the volume ratio of the supernatant to the eutectic solvent is 1:0.5; the extraction temperature is 45℃, and the extraction time is 45 min.
[0056] Comparative Example 4
[0057] The Cyclobalanopsis gaber leaves are washed, dried and crushed, and then sieved through a 60-mesh sieve; the crushed Cyclobalanopsis gaber leaves are then dispersed in a citric acid-phosphate buffer, the pH of the enzymatic system is controlled at 5.5, and then a compound enzyme is added for enzymolysis; a eutectic solvent is added to the supernatant after the enzymolysis; and the solution after the extraction is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, to obtain the Cyclobalanopsis gaber extract.
[0058] The mass ratio of the Cyclobalanopsis gaber leaves, the compound enzyme and the citric acid-phosphate buffer is 1:0.03:15.
[0059] The compound enzyme is composed of cellulase and pectinase at a mass ratio of 1:1; the enzymolysis temperature is 50℃, and the enzymolysis time is 90 min.
[0060] The eutectic solvent is composed of choline chloride and erythritol at a molar ratio of 1:1; the volume ratio of the supernatant to the eutectic solvent is 1:0.5; the extraction temperature is 45℃, and the extraction time is 45 min.
[0061] Comparative Example 5
[0062] The Cyclobalanopsis gaber leaves are washed, dried and crushed, and then sieved through a 60-mesh sieve; the crushed Cyclobalanopsis gaber leaves are then dispersed in a citric acid-phosphate buffer, the pH of the enzymatic system is controlled at 5.5, and then a compound enzyme is added for enzymolysis; a eutectic solvent is added to the supernatant after the enzymolysis; and the solution after the extraction is separated by a macroporous resin, eluted by ethanol and concentrated by rotary evaporation, to obtain the Cyclobalanopsis gaber extract.
[0063] The mass ratio of the Cyclocarya paliurus leaf, the complex enzyme and the citric acid-phosphate buffer is 1:0.03:15.
[0064] The complex enzyme is composed of cellulase and pectinase at a mass ratio of 1:1; the enzymolysis temperature is 50℃, and the enzymolysis time is 90 min.
[0065] The deep eutectic solvent is composed of choline chloride and tartaric acid at a molar ratio of 1:1; the volume ratio of the supernatant to the deep eutectic solvent is 1:0.5; the extraction temperature is 45℃, and the extraction time is 45 min.
[0066] The effective components in the extracts of Example 1 and Comparative Examples 1-5 are detected by HPLC fingerprint spectrum method, and the test results are shown in Table 1.
[0067] Table 1: Test results of effective components in extracts (mg / g)
[0068]
[0069] As can be seen from the test results of Example 1 and Comparative Examples 1-5 in Table 1, in the presence of an alternating magnetic field, the tartaric acid and erythritol in the deep eutectic solvent of the present application can achieve a synergistic promotion effect, which significantly improves the extraction rate of effective components such as neochlorogenic acid, chlorogenic acid, quercetin-3-O-glucuronide, quercitrin and afzelin in Cyclocarya paliurus. The related mechanism is as follows: first, because the alternating magnetic field can make the tartaric acid, erythritol and choline chloride in the deep eutectic solvent produce alternating orientation and enhance their polarizability, this high-frequency molecular shock can break some weak hydrogen bonds and form more stable and denser hydrogen bond networks in a new conformation, which dynamically optimizes the synergistic structure between tartaric acid and erythritol (i.e. complex hydrogen bond entanglement formed by hydroxyl and carboxyl groups), thereby forming more “temporary association bodies” and enhancing the complexing and wrapping capacity of the solvent for polyphenol and flavonoid molecules in Cyclocarya paliurus; second, the alternating magnetic field introduces micro-perturbation and micro-turbulence in the liquid, which can induce the solvent to have enhanced flowability, reduced viscosity and improved diffusion coefficient in the system involving erythritol and tartaric acid, thereby improving the penetration, cleaning and infiltration capacity of the solvent into plant cell walls; finally, the alternating magnetic field can also induce some intracellular components that are not completely released by enzymolysis to be further released under the dynamic action of the solvent, at this time, the weak acid property of tartaric acid helps to maintain the extracellular-intracellular pH gradient and accelerate the diffusion of the components outside, while the multiple hydroxyl sites of erythritol can dynamically bind the free components to form a complex state, preventing them from being oxidized and aggregated.
[0070] While the embodiments of the application have been shown and described, it is to be understood that the application is not limited to the details of the embodiments described, since the scope of the expenditure of the application is defined with respect to the appended claims and their equivalents.
Claims
1. A method for extracting Cyclobalanopsis glauca, characterized by, The method comprises the following steps: S1: washing, drying and crushing the Cyclobalanopsis glauca leaves; S2: dispersing the crushed Cyclobalanopsis glauca leaves in a citric acid-phosphate buffer, and adding a compound enzyme for enzymolysis; S3: adding a deep eutectic solvent to the supernatant after enzymolysis, and extracting under the action of an alternating magnetic field; S4: separating the extracted solution through a macroporous resin, eluting with ethanol, and concentrating by rotary evaporation to obtain the Cyclobalanopsis glauca extract.
2. The extraction method of the Spondias mombin according to claim 1, characterized by, The mass ratio of the Cyclobalanopsis glauca leaves, the compound enzyme and the citric acid-phosphate buffer is 1:0.01-0.05:10-20.
3. The extraction method of the Adenanthera pavonina according to claim 2 or 3, characterized in that, The compound enzyme is composed of cellulase and pectinase at a mass ratio of 2:1-4.
4. The extraction method of the Spondias mombin according to claim 2 or 3, characterized by, The enzymolysis temperature is 40-60℃, and the enzymolysis time is 60-120 min.
5. The extraction method of the Spondias mombin according to claim 1, characterized by, The deep eutectic solvent is composed of choline chloride, tartaric acid and erythritol at a molar ratio of 1:0.4-0.6:0.4-0.6; and the volume ratio of the supernatant to the deep eutectic solvent is 1:0.1-1.
6. The extraction method of the Spondias mombin according to claim 1, characterized by, The frequency of the alternating magnetic field is 40-60 Hz, and the intensity is 0.2-0.4 T.
7. The extraction method of the Spondias mombin according to claim 1, characterized by, The extraction temperature is 40-50℃, and the extraction time is 30-60 min.