Method for simultaneously extracting flavonoid, phenol and polysaccharide traditional Chinese medicine effective components by ultrasonic-assisted nanoemulsion

Abstract

The invention relates to the technical field of extraction and separation of effective components of traditional Chinese medicines, and discloses a method for simultaneously extracting effective components of traditional Chinese medicines such as flavonoids, phenols and polysaccharides by using an ultrasonic-assisted nanoemulsion. Pouring the filtered traditional Chinese medicine powder, continuously stirring for 20 minutes through an ultrasonic cavitation technology and magnetic stirring, centrifuging for 15-20 minutes, and separating a solid phase from a liquid phase after centrifuging, so as to obtain the separated liquid phase, namely the nano-emulsion containing the effective components. According to the method for extracting flavone, phenols and polysaccharides from traditional Chinese medicines by ultrasonic-assisted nanoemulsion, an oil phase and a water phase are crushed to form nanoscale liquid drops, so that dispersion of the emulsion drops can be spontaneously generated, and physical damage of effective components of the traditional Chinese medicines to the medicines due to high temperature or high pressure and a mechanical process can be avoided or relieved; the prepared nano-emulsion not only improves the loading rate, the long-term stability, the dilution stability and the light stability of the material, but also reduces the toxicity of the surfactant.

Description

Technical Field

[0001] This invention relates to the field of extraction and separation technology of effective components of traditional Chinese medicine, specifically a method for simultaneously extracting flavonoids, phenols and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion. Background Technology

[0002] Traditional Chinese medicine (TCM) is effective in treating diseases, with low toxicity and few side effects, making it relatively safe and aligning with modern concepts of disease prevention and treatment. However, the immaturity of TCM production technology and processes remains a major long-standing problem in my country. The research and utilization of TCM largely depend on the extraction and separation of its active ingredients. Given the complex composition of TCM and natural products, extraction and analysis methods are crucial. Traditional extraction processes such as decoction, reflux, soaking, and percolation suffer from drawbacks including significant losses, long cycles, numerous steps, and low extraction rates. Even with rapid advancements in TCM production technology and processes, the extraction efficiency and content of TCM herbs remain low, failing to meet industrial production needs and resulting in waste. With the deepening of modern TCM research, extraction technologies for the active ingredients of traditional TCM are constantly being innovated to improve extraction efficiency and bioavailability.

[0003] "Gui Shi Wei" (Ten Delicacies of Cinnamon) is based on the "Standards for Authentic Medicinal Materials of the China Association of Traditional Chinese Medicine" and adheres to the principle of "three generations of herbal medicine and a century of history." It has the advantages of "high quality, good efficacy, high popularity, and profound cultural heritage." The ingredients are: cinnamon (including cassia twigs), monk fruit, star anise, Guangxi turmeric (including cinnamon turmeric), longan pulp, sophora root, chicken blood vine, chicken bone grass, thorny lily, and earthworm. These are characteristic authentic medicinal materials from Guangxi.

[0004] This category of medicinal herbs is rich in flavonoids, phenols, and polysaccharides. Chicken blood vine, chicken bone grass, and monk fruit are commonly used traditional Chinese medicines, widely applied in TCM to treat diseases such as blood circulation disorders and immune regulation. Chicken blood vine contains abundant polyphenols and flavonoids, exhibiting excellent antioxidant, liver-protective, and anti-inflammatory effects. Monk fruit is an important source of natural sweeteners; its flavonoids and polysaccharides possess antioxidant, hypoglycemic, and anti-inflammatory functions. Chicken bone grass contains flavonoids, saponins, and other active ingredients, exhibiting excellent antioxidant, anti-inflammatory, and lipid-regulating properties. However, existing extraction methods mostly employ traditional soaking and decoction techniques, which suffer from low extraction efficiency, high solvent consumption, long extraction times, and complex extracted components, limiting their large-scale application and the enhancement of their clinical value.

[0005] In recent years, the application of nanotechnology in the extraction of traditional Chinese medicine has received widespread attention. Nanoemulsions, as emulsion systems with extremely small particle sizes and high surface areas, can effectively improve the solubility and bioavailability of traditional Chinese medicine components. Through a rational combination of emulsifiers and co-emulsifiers, and the application of ultrasonic cavitation technology, nanoemulsions can promote the rapid release of the active ingredients of the drug, thereby improving extraction efficiency. Existing extraction technologies have the following problems: 1. Traditional extraction methods (such as alcohol extraction, hot water extraction, and organic solvent extraction) rely on the polarity and diffusion ability of the solvent, making it difficult to fully release the effective components in the medicinal materials, especially hydrophobic components (such as flavonoids and phenols).

[0006] 2. Traditional extraction methods usually require long-term heating or the use of large amounts of solvent, which is energy-intensive and may damage the target components.

[0007] 3. Traditional extraction methods typically have a wide extraction range, making it difficult to selectively extract specific components, and the extract often contains many impurities. Summary of the Invention

[0008] (a) Technical problems to be solved

[0009] To address the shortcomings of existing technologies, this invention provides a method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsions. This method offers advantages such as improved material loading rate, long-term stability, dilution stability, and photostability, while solving the problems of low effective ingredient content and high impurity levels.

[0010] (ii) Technical issues

[0011] To achieve the above objectives, the present invention provides the following technical solution: a method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion, comprising the following steps: (1) Use ultra-micro pulverization technology to pulverize Chinese medicine into dust with a particle size of 10~50μm and then filter it.

[0012] (2) Prepare the oil phase with castor oil and ethyl acetate in a 1:1 ratio, use sorbitan monopalmitate or polyoxyethylene stearate as emulsifier, use propylene glycol and glycerin in a 2:1 ratio as co-emulsifier, and use deionized water as the aqueous phase. Prepare the extractant with the ratio of oil phase: emulsifier: co-emulsifier: aqueous phase in a 2:3:1:1 ratio.

[0013] (3) After mixing the extractant from (2) with magnetic stirring (12000~13000rpm), pour it into the filtered Chinese medicine powder and continue stirring with magnetic stirring for 20min using ultrasonic cavitation technology, then centrifuge for 15~20min.

[0014] (4) After centrifugation, the solid phase and liquid phase are separated. The separated liquid phase is the nanoemulsion containing the effective ingredients.

[0015] Traditional Chinese medicine is processed into powder for extraction using ultra-micro pulverization technology. Nanoemulsion is prepared by mixing emulsifier and co-emulsifier at room temperature using a magnetic stirrer for a certain time (30-60 min), then slowly adding the oil phase and stirring until homogeneous. A small amount of aqueous phase is then slowly added dropwise to the mixture until it changes from a gel state to a clear, transparent liquid state through stirring. Aqueous phase is then added uniformly until the total volume is reached, yielding the nanoemulsion. The powder to be extracted is mixed with the nanoemulsion (the temperature can be appropriately increased at this point). The mixture is then homogeneously mixed using ultrasonic cavitation technology and magnetic stirring, followed by centrifugation (12000-13000 rpm, stirring for 20 min, centrifugation for 15-20 min). After centrifugation, the solid and liquid phases are separated; the separated liquid phase is the nanoemulsion containing the active ingredients.

[0016] In the above steps, when processing the raw materials using ultrafine grinding technology, firstly, the dried raw materials are chopped into small pieces of about 5 cm. Then, an air jet mill (or other suitable equipment) is used for ultrafine grinding, adjusting the equipment parameters to ensure that the particle size after grinding is within the range of 10~50 μm. During the grinding process, a cooling device is used to control the temperature to not exceed 15°C to prevent the degradation of heat-sensitive components. After grinding, a sieving device is used to remove particles that do not meet the size requirements, retaining the powder with uniform particle size for subsequent processing.

[0017] In the above steps, after the powder to be extracted is mixed with the nanoemulsion, ultrasonic cavitation technology and magnetic stirring are used to further promote the extraction of active ingredients. Specifically, the powder to be extracted is mixed with the prepared nanoemulsion mixture in a certain proportion, and the temperature is appropriately increased to enhance the solubility of the substances. Subsequently, the mixture is subjected to ultrasonic cavitation treatment using an ultrasonic device, with the ultrasonic power set to 300-500 W, the frequency to 20-40 kHz, and the treatment time to 15-30 min. The cavitation effect of ultrasound forms microbubbles that rapidly burst, generating strong shear force and microjets, significantly improving the dissolution and emulsion stability of the active ingredients. Simultaneously, a magnetic stirrer is used to continuously stir the mixture to ensure uniform mixing and promote full reaction of the materials.

[0018] The solid phase after centrifugation can be re-extracted 1 to 3 times by adding a trace amount of nanoemulsion.

[0019] Preferably, the traditional Chinese medicine is pulverized using ultra-micro technology to a particle size of 10-50 μm, and the traditional Chinese medicine dust of 10-50 μm is screened out for later use.

[0020] Preferably, the oil phase is prepared by mixing castor oil and ethyl acetate in a 1:1 ratio.

[0021] Preferably, sorbitan monopalmitate or polyoxyethylene stearate is used as an emulsifier.

[0022] Preferably, the ratio of emulsifier to co-emulsifier is 3:1, and the ratio of oil phase: emulsifier: co-emulsifier: aqueous phase of extractant is 2:3:1:1.

[0023] Preferably, deionized water is used as the aqueous phase.

[0024] Preferably, the filtered Chinese medicine powder is further stirred by ultrasonic cavitation technology and magnetic stirring, and then centrifuged.

[0025] Preferably, the solid waste is repeatedly extracted.

[0026] Compared with existing technologies, this invention provides a method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsions, which has the following beneficial effects: (1) The microemulsion method is used to extract the effective components of traditional Chinese medicine into nanoemulsions, which can avoid the problems of excessive loss of effective components, insufficient concentration, many impurities, a small amount of fat solubility and large-scale decomposition of effective components caused by traditional extraction methods.

[0027] (2) The microemulsion method is used to extract the effective components of traditional Chinese medicine into nanoemulsions. Only solid-liquid separation is required to obtain the effective components of traditional Chinese medicine, which reduces the process flow of traditional Chinese medicine extraction and separation.

[0028] (3) The effective components of traditional Chinese medicine made from nanoemulsions have application advantages, and the purification rate of nanoemulsions under ultrasound assistance is high.

[0029] (4) In the ultrasonic-assisted extraction process of traditional Chinese medicine, ultrafine grinding technology can avoid local overheating and other phenomena, and the grinding speed is relatively fast. Therefore, it can maximize the preservation of the biological activity and various nutrients of the powder, reduce the loss of effective ingredients, and is conducive to the development and preparation of high-quality products. Therefore, ultrafine grinding technology is not only suitable for grinding fibrous materials (especially for grinding materials containing aromatic and volatile components), but also can be used for medium, low and ultra-low temperature grinding according to the needs of different materials, so as to achieve better product effects according to the material properties and processing requirements, thereby increasing the adsorption and solubility of the product. Ultrafine grinding is carried out in a closed system, which avoids the pollution of the surrounding environment by micropowder and prevents dust in the air from contaminating the product. The application of this technology in food and health care products can control the contamination of microorganisms and dust. At the same time, since ultrafine grinding is a purely physical process, it will not mix in other impurities, which also makes the processed Chinese herbal medicines pure natural, ensuring the integrity and safety of the raw material components.

[0030] (5) In the ultrasonic-assisted extraction process of traditional Chinese medicine, ultrasonic cavitation technology generates strong destructive force, which breaks down the oil phase and water phase into nano-sized droplets, enabling the spontaneous generation of emulsion droplets. This can avoid or reduce the physical damage to the active ingredients of traditional Chinese medicine caused by high temperature, high pressure and mechanical processes. The nanoemulsion prepared by ultrasonic cavitation technology not only improves the loading rate, long-term stability, dilution stability and light stability of the material, but also reduces the toxicity of surfactants. Attached Figure Description

[0031] Figure 1 This is a flowchart of the ultrasonic nanoemulsion method for simultaneously extracting multiple components of traditional Chinese medicine according to the present invention; Figure 2 This is a roadmap for the ultrasonic nanoemulsion refinement technique of the present invention. Detailed Implementation

[0032] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0033] Please see Figure 1-2 A method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion includes the following steps: (1) Use ultra-micro pulverization technology to pulverize Chinese medicine into dust with a particle size of 10~50μm and then filter it.

[0034] (2) Prepare the oil phase with castor oil and ethyl acetate in a 1:1 ratio, use sorbitan monopalmitate or polyoxyethylene stearate as emulsifier, use propylene glycol and glycerin in a 2:1 ratio as co-emulsifier, and use deionized water as the aqueous phase. Prepare the extractant with the ratio of oil phase: emulsifier: co-emulsifier: aqueous phase in a 2:3:1:1 ratio.

[0035] (3) After mixing the extractant from (2) with magnetic stirring (12000~13000rpm), pour it into the filtered Chinese medicine powder and continue stirring with magnetic stirring for 20min using ultrasonic cavitation technology, then centrifuge for 15~20min.

[0036] (4) After centrifugation, the solid phase and liquid phase are separated. The separated liquid phase is the nanoemulsion containing the effective ingredients.

[0037] Traditional Chinese medicine (TCM) is pulverized using ultra-micro technology to a particle size of 10-50 μm, and TCM dust of 10-50 μm is screened out for later use. The TCM is then pulverized into a powder to be extracted using ultra-micro pulverization technology. Nanoemulsion is prepared by mixing emulsifiers and co-emulsifiers at room temperature using a magnetic stirrer for a certain time (30-60 min). The oil phase is slowly added and stirred until homogeneous. A small amount of aqueous phase is then slowly added dropwise to the mixture until it changes from a gel state to a clear, transparent liquid state through stirring. The aqueous phase is then added uniformly until the total volume is reached, yielding the nanoemulsion. The powder to be extracted is mixed with the nanoemulsion (the temperature can be appropriately increased at this point). The mixture is then homogeneously mixed using ultrasonic cavitation technology and magnetic stirring, followed by centrifugation (12000-13000 rpm, stirring for 20 min, centrifugation for 15-20 min). After centrifugation, the solid and liquid phases are separated; the separated liquid phase is the nanoemulsion containing the active ingredients.

[0038] In the above steps, when processing the raw materials using ultrafine grinding technology, firstly, the dried raw materials are chopped into small pieces of about 5 cm. Then, an air jet mill (or other suitable equipment) is used for ultrafine grinding, adjusting the equipment parameters to ensure that the particle size after grinding is within the range of 10~50 μm. During the grinding process, a cooling device is used to control the temperature to not exceed 15°C to prevent the degradation of heat-sensitive components. After grinding, a sieving device is used to remove particles that do not meet the size requirements, retaining the powder with uniform particle size for subsequent processing.

[0039] In the above steps, after the powder to be extracted is mixed with the nanoemulsion, ultrasonic cavitation technology and magnetic stirring are used to further promote the extraction of active ingredients. Specifically, the powder to be extracted is mixed with the prepared nanoemulsion mixture in a certain proportion, and the temperature is appropriately increased to enhance the solubility of the substances. Subsequently, the mixture is subjected to ultrasonic cavitation treatment using an ultrasonic device, with the ultrasonic power set to 300-500 W, the frequency to 20-40 kHz, and the treatment time to 15-30 min. The cavitation effect of ultrasound forms microbubbles that rapidly burst, generating strong shear force and microjets, significantly improving the dissolution and emulsion stability of the active ingredients. Simultaneously, a magnetic stirrer is used to continuously stir the mixture to ensure uniform mixing and promote full reaction of the materials. The solid phase after centrifugation can be re-extracted 1-3 times by adding a trace amount of nanoemulsion.

[0040] The oil phase was prepared using a 1:1 ratio of castor oil and ethyl acetate. Sorbitol monopalmitate or polyoxyethylene stearate was used as the emulsifier. The ratio of emulsifier to co-emulsifier was 3:1, and the ratio of oil phase:emulsifier:co-emulsifier:aqueous phase of the extract was 2:3:1:1. Deionized water was used as the aqueous phase. The filtered herbal powder was further stirred using ultrasonic cavitation and magnetic stirring, followed by centrifugation. The solid waste was repeatedly extracted.

[0041]

[0042] Comparison of this invention with traditional extraction methods:

[0043] Traditional extraction methods (such as alcohol extraction, hot water extraction, and organic solvent extraction) rely on the polarity and diffusion capacity of the solvent, making it difficult to fully release the active ingredients in medicinal materials, especially hydrophobic components (such as flavonoids and phenols). Traditional extraction methods typically require prolonged heating or the use of large amounts of solvent, resulting in high energy consumption and potential damage to the target components. Furthermore, traditional extraction methods generally have a broad extraction range, making it difficult to selectively extract specific components, and the extract often contains many impurities.

[0044] Ultrasonic extraction is performed at room temperature or low temperature, avoiding the damage to heat-sensitive components (such as flavonoids, phenols, and polysaccharides) caused by high temperatures in traditional methods. Through the mechanical vibration and cavitation effect of ultrasound, the cell walls of medicinal materials are disrupted, accelerating the release of target components and significantly shortening the extraction time. This invention, through the unique formulation of nanoemulsions (surfactant + oil phase), significantly improves the solubilization ability for hydrophobic components. The nano-sized particles of the nanoemulsions can quickly penetrate into the cells of medicinal materials, releasing target components and greatly improving extraction efficiency, thus solving the problem of low extraction efficiency in traditional methods. This invention achieves selective extraction of specific components (such as flavonoids, phenols, and polysaccharides) by adjusting the formulation of the nanoemulsions (such as surfactant type and oil phase selection). Furthermore, the extract obtained by combining nanoemulsions with ultrasonic extraction has fewer impurities and higher purity, reducing subsequent purification steps.

[0045] The table below shows the experimental comparison data of the effective component extraction rates of the traditional method and the method of the present invention: (wherein, the ultrasound-assisted nanoemulsion method is as described in the present invention; alcohol extraction or water decoction method: using 70% ethanol or water, heated and refluxed for 2 hours; hot water extraction method: water decoction extraction twice, 1.5 hours each time; organic solvent extraction method: using ethyl acetate or methanol, heated and refluxed for 2 hours.)

[0046] Traditional extraction methods require different techniques for each active ingredient. For example, flavonoids are often extracted using alcohol extraction or water decoction, polysaccharides using hot water extraction, and phenols using organic solvent extraction. These traditional methods often suffer from low extraction efficiency due to long extraction times, high levels of impurities in the extract, and the potential for chemical bond breakage and oxidation at high temperatures. However, ultrasound-assisted nanoemulsion extraction can simultaneously extract flavonoids, phenols, and polysaccharides from traditional Chinese medicine, offering significant improvements in extraction efficiency, component integrity, selectivity, and environmental friendliness.

[0047] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion, characterized in that... Includes the following steps: (1) Use ultra-micro pulverization technology to pulverize Chinese medicine into dust with a particle size of 10~50μm and then filter it; (2) Prepare the oil phase with castor oil and ethyl acetate in a 1:1 ratio, use sorbitan monopalmitate or polyoxyethylene stearate as emulsifier, use propylene glycol and glycerin in a 2:1 ratio as co-emulsifier, wherein the ratio of emulsifier to co-emulsifier is 3:1, use deionized water as the aqueous phase, and prepare the extractant with the ratio of oil phase: emulsifier: co-emulsifier: aqueous phase in a 2:3:1:1 ratio; (3) After mixing the extractant from (2) with magnetic stirring (12000~13000rpm), pour it into the filtered Chinese medicine powder and continue stirring with magnetic stirring for 20min using ultrasonic cavitation technology, then centrifuge for 15~20min. (4) After centrifugation, the solid phase and liquid phase are separated. The separated liquid phase is the nanoemulsion containing the effective ingredients.

2. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: Traditional Chinese medicine is pulverized using ultra-micro technology to a particle size of 10-50 μm, and the 10-50 μm powder is screened out for later use.

3. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: Prepare the oil phase using a 1:1 ratio of castor oil and ethyl acetate.

4. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: Use dehydrated sorbitan monopalmitate or polyoxyethylene stearate as emulsifiers.

5. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: The ratio of emulsifier to co-emulsifier is 3:1, and the ratio of oil phase: emulsifier: co-emulsifier: aqueous phase of extractant is 2:3:1:

1.

6. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: Deionized water is used as the aqueous phase.

7. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: The filtered Chinese medicine powder is further stirred using ultrasonic cavitation technology and magnetic stirring, and then centrifuged.

8. The method for simultaneously extracting flavonoids, phenols, and polysaccharides from traditional Chinese medicine using ultrasound-assisted nanoemulsion according to claim 1, characterized in that: The solid waste was repeatedly extracted.

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