Process optimization method for green extraction of hemsley rockvine root flavonoid glycoside through cooperation of ultrasonic and surfactant

Abstract

The invention relates to the technical field of traditional Chinese medicine extraction, in particular to a process optimization method for green extraction of hemsley rockvine root flavonoid glycoside through ultrasonic cooperation with a surfactant. The method comprises the following steps: optimizing process parameters for extracting isoorientin, isoquercitrin, kaempferol-3-O-rutinoside and astragalin from radix tetrastigme tuberous roots by using an ultrasonic-assisted lauryl sodium sulfate aqueous solution micelle medium; influences of the type and concentration of a surfactant, a material-liquid ratio, ultrasonic power, ultrasonic time, ultrasonic times and ultrasonic temperature on target components are investigated, and optimal extraction process parameters are determined: the concentration of sodium dodecyl sulfate (SDS) is 35mg / mL, the ultrasonic power is 100W, and the ultrasonic temperature is 50 DEG C. According to the process, a traditional organic solvent is replaced by the surfactant, so that the extraction efficiency is improved; a feasible scheme is provided for green extraction of active ingredients of radix tetrastigme, and industrial amplification feasibility can be further explored subsequently.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine extraction, in particular to a process optimization method for green extraction of trifoliate green flavonoid glycosides by using ultrasound and surfactant in combination. Background Art

[0002] As a traditional Chinese medicinal material, Sanyeqing has multiple pharmacological activities, among which flavonoid glycoside component is one of its main active ingredients. In the existing technology, the extraction of Sanyeqing flavonoid glycosides mostly adopts the traditional solvent extraction method, which has problems such as large solvent consumption, low extraction efficiency, and serious environmental pollution. In recent years, the application of ultrasound-assisted extraction and surfactants has provided new ideas for the extraction of traditional Chinese medicine. Ultrasonic waves can produce strong cavitation and vibration effects, destroy plant cell walls, and improve extraction efficiency; surfactants can reduce the surface tension of the solution and enhance the solubility of the solute in the solvent, thereby improving the extraction effect.

[0003] However, the current process of ultrasound-assisted surfactant extraction of trifoliate green flavonoid glycosides still has problems such as unreasonable parameter settings and unclear extraction mechanism, which limits its industrial application. Summary of the Invention

[0004] The purpose of the present invention is to provide a process optimization method for the green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactants, so as to solve the problems in the existing technology of the extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactants, such as unreasonable parameter settings and unclear extraction mechanism, which limit its industrial application.

[0005] To achieve the above object, the present invention provides a process optimization method for the green extraction of flavonoid glycosides of trifoliate green by using ultrasound and surfactant, and the process optimization method for the green extraction of flavonoid glycosides of trifoliate green by using ultrasound and surfactant comprises the following steps:

[0006] Step S1: Accurately weigh 1 g of Tripterygium wilfordii powder and mix it evenly with 5% aqueous solutions of 9 surfactants at a solid-liquid ratio of 1:10 (g / mL). Extraction was performed under the same ultrasonic conditions. Three parallel experiments were conducted in each group to investigate the saturated solubility of ISO, ISQ, K3R, and AST in Tripterygium wilfordii roots with different surfactant aqueous solutions and the extraction rate of target components in Tripterygium wilfordii roots by ultrasonic-assisted extraction, so as to determine the optimal surfactant type.

[0007] Step S2: Screening out 6 main factors that affect the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii, conducting experiments on individual factors, and conducting 3 parallel experiments in each group, and then screening out the final factor that affects the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii;

[0008] Step S3: Box-Behnken Design was used to further optimize the extraction process conditions of ISO, ISQ, K3R, and AST from the root tubers of Tripterygium wilfordii using ultrasound-assisted micellar medium, and four influencing factors were selected as independent variables;

[0009] Step S4: performing auxiliary extraction in combination with a deep eutectic solvent, recording and comparing the extraction effects under different solvent systems, so as to minimize the amount of organic solvent used and achieve friendly extraction;

[0010] Step S5: Molecular dynamics simulation software is used to simulate the interaction process between the optimal surfactant type and flavonoid glycosides in Tripterygium wilfordii, and the variables of the final factor and the four influencing factors are controlled. At the same time, FT-IR analysis, SEM / TEM observation and Fick's second law model are combined to comprehensively explain the synergistic mechanism of surfactants on the transmembrane transport of flavonoid glycosides, and finally complete the optimization of the Tripterygium wilfordii flavonoid glycoside process.

[0011] Among them, in step S1, the nine surfactants are sodium lauryl sulfate, sodium lauryl sulfonate, sodium lauryl sulfate, cocamidopropyl betaine, dodecyl dimethyl betaine, hexadecyl trimethyl ammonium chloride, alkyl glucoside, sucrose fatty acid ester and polyvinyl alcohol, and the extraction power is set to 360W, the ultrasonic temperature is below 30°C, and the extraction is 20min.

[0012] Among them, in step S2, the six main factors affecting the extraction rates of ISO, ISQ, K3R and AST in the tuberous roots of Tripterygium wilfordii are ultrasonic power, material-liquid ratio, number of ultrasonic times, ultrasonic time, surfactant concentration and ultrasonic temperature.

[0013] Among them, in step S3, the four influencing factors and independent variables are ultrasonic power (X1: 252-360 W), ultrasonic temperature (X2: 30-50 ° C), SDS concentration (X3: 10-60 mg / mL) and material-liquid ratio (X4: 1:10-1:20 g / mL).

[0014] Wherein, in step S3, the deep eutectic solvent is one of a choline lactic acid-water system or supercritical CO2.

[0015] Wherein, in step S4, the molecular dynamics simulation software is GROMACS.

[0016] Among them, in step S4, the specific analysis content of the FT-IR analysis is to compare the changes in characteristic peaks of the cell wall components of Trifoliate Green before and after extraction, and to reveal the destruction mechanism of the cell wall by the surfactant.

[0017] Wherein, in step S4, the SEM / TEM observation is to observe the morphological changes of plant cells before and after ultrasonic treatment using a scanning or transmission electron microscope to clarify the effects of the cavitation effect and the synergistic effect of the surfactant on the permeability of the cell wall.

[0018] Wherein, in step S4, the Fick's second law model is specifically used to calculate the diffusion coefficient of flavonoid glycosides, and to illustrate the mechanism by which surfactants enhance the mass transfer rate.

[0019] The invention discloses a process optimization method for green extraction of flavonoid glycosides from Tripterygium wilfordii using ultrasound-assisted surfactant. The process parameters for extracting isoorientin, isoquercetin, kaempferol-3-O-rutinoside and astragaloside from Tripterygium wilfordii root tubers using an ultrasound-assisted sodium dodecyl sulfate aqueous solution micellar medium are optimized, the effects of the type and concentration of the surfactant, the material-liquid ratio, the ultrasonic power, the ultrasonic time, the number of ultrasonic times and the ultrasonic temperature on the target components are investigated, and the optimal extraction process parameters are determined as follows: sodium dodecyl sulfate (SDS) concentration: 35 mg / mL, the ultrasonic power: 100 W, and the ultrasonic temperature: 50°C. Under the optimal extraction process conditions, the average extraction rates of isoorientin, isoquercetin, kaempferol-3-O-rutinoside and astragaloside are 5.856 μg / g, 15.296 μg / g, 17.474 μg / g and 15.683 μg / g, respectively. The differences in the extraction of various components are related to the polar micelle solubilization ability of flavonoid glycosides. This process replaces traditional organic solvents with surfactants, providing a feasible solution for the green extraction of the active ingredients of Tripterygium wilfordii. The feasibility of industrial scale-up can be further explored in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only 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.

[0021] Figure 1 This is a flow chart of the steps of the process optimization method for green extraction of trifoliate green flavonoid glycosides by using ultrasound and surfactant provided by the present invention.

[0022] Figure 2 The present invention provides a curve diagram showing the influence of surfactant concentration on ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactant provided by the present invention.

[0023] Figure 3The present invention provides a curve diagram showing the effects of the material-liquid ratio on ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides using ultrasound-assisted surfactants.

[0024] Figure 4 The present invention provides a curve diagram showing the effect of ultrasound time on ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides using ultrasound-assisted surfactants.

[0025] Figure 5 The present invention provides a curve diagram showing the relationship between the number of ultrasound waves and ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides by using ultrasound and surfactants.

[0026] Figure 6 The present invention provides a curve diagram showing the relationship between ultrasonic power, ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides by using ultrasound and surfactants.

[0027] Figure 7 The present invention provides a curve diagram showing the effects of ultrasonic temperature on ISO, ISQ, AST and AST extraction rate in the process optimization method for green extraction of flavonoid glycosides of trifoliate green using ultrasonic synergistic surfactants.

[0028] Figure 8 This is a coefficient of variation diagram of the process optimization method for green extraction of trifoliate green flavonoid glycosides using ultrasound-assisted surfactants provided by the present invention.

[0029] Figure 9 It is a response surface diagram of the influence of different interaction factors on ISO extraction rate in the process optimization method of green extraction of trifoliate green flavonoid glycosides by ultrasound synergistic surfactant provided by the present invention.

[0030] Figure 10 It is a response surface diagram of the influence of different interaction factors on the ISQ extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound synergistic surfactant provided by the present invention.

[0031] Figure 11 It is a response surface diagram of the effects of different interaction factors on the K3R extraction rate in the process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactant provided by the present invention.

[0032] Figure 12 It is a response surface diagram of the influence of different interactive factors on the AST extraction rate in the process optimization method of the ultrasound-assisted surfactant green extraction of flavonoid glycosides provided by the present invention. DETAILED DESCRIPTION

[0033] The following describes embodiments of the present invention in detail, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and are not to be construed as limiting the present invention.

[0034] See also Figures 1 to 12 The present invention provides a process optimization method for green extraction of flavonoid glycosides from Tripterygium wilfordii using ultrasound-assisted surfactants. The process optimization method for green extraction of flavonoid glycosides from Tripterygium wilfordii using ultrasound-assisted surfactants comprises the following steps:

[0035] Step S1: Accurately weigh 1 g of Tripterygium wilfordii powder and mix it evenly with 5% aqueous solutions of 9 surfactants at a solid-liquid ratio of 1:10 (g / mL). Extraction was performed under the same ultrasonic conditions. Three parallel experiments were conducted in each group to investigate the saturated solubility of ISO, ISQ, K3R, and AST in Tripterygium wilfordii roots with different surfactant aqueous solutions and the extraction rate of target components in Tripterygium wilfordii roots by ultrasonic-assisted extraction, so as to determine the optimal surfactant type.

[0036] Step S2: Screening out 6 main factors that affect the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii, conducting experiments on individual factors, and conducting 3 parallel experiments in each group, and then screening out the final factor that affects the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii;

[0037] Step S3: Box-Behnken Design was used to further optimize the extraction process conditions of ISO, ISQ, K3R, and AST from the root tubers of Tripterygium wilfordii using ultrasound-assisted micellar medium, and four influencing factors were selected as independent variables;

[0038] Step S4: performing auxiliary extraction in combination with a deep eutectic solvent, recording and comparing the extraction effects under different solvent systems, so as to minimize the amount of organic solvent used and achieve friendly extraction;

[0039] Step S5: Molecular dynamics simulation software is used to simulate the interaction process between the optimal surfactant type and flavonoid glycosides in Tripterygium wilfordii, and the variables of the final factor and the four influencing factors are controlled. At the same time, FT-IR analysis, SEM / TEM observation and Fick's second law model are combined to comprehensively explain the synergistic mechanism of surfactants on the transmembrane transport of flavonoid glycosides, and finally complete the optimization of the Tripterygium wilfordii flavonoid glycoside process.

[0040] In this embodiment, HPLC analysis of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii was performed using an Agilent ZORBAX SB-C18 (4.6×250 mm) column with a mobile phase consisting of acetonitrile (A)-0.1% formic acid water (containing 5% methanol) (B), a flow rate of 1.0 mL·min-1, an injection volume of 10 μL, a detection wavelength of 265 nm, a column temperature of 25°C, and three replicates per group. Gradient elution program: 0 min, 10% A, 90% B; 30 min, 20% A, 80% B; 35 min, 20% A, 80% B; 45 min, 30% A, 70% B; 50 min, 80% A, 20% B; 55 min, 80% A, 20% B.

[0041] In this embodiment, the formula for calculating the extraction rate of ISO, ISQ, K3R, and AST in the root tuber of Tripterygium wilfordii is:

[0042]

[0043] Wherein, C is the concentration of the substance being measured in the sample solution (μg / mL), V is the volume of the sample solution (mL), and M is the dry weight of the trifoliate green powder (g).

[0044] Furthermore, in step S1, the nine surfactants are sodium lauryl sulfate, sodium lauryl sulfonate, sodium lauryl sulfate, cocamidopropyl betaine, dodecyl dimethyl betaine, hexadecyl trimethyl ammonium chloride, alkyl glucoside, sucrose fatty acid ester and polyvinyl alcohol, and the extraction power is set to 360 W, the ultrasonic temperature is below 30° C., and the extraction time is 20 min.

[0045] Furthermore, in step S2, the six main factors affecting the extraction rates of ISO, ISQ, K3R and AST in the tuberous roots of Tripterygium wilfordii are ultrasonic power, solid-liquid ratio, ultrasonic times, ultrasonic time, surfactant concentration and ultrasonic temperature.

[0046] Furthermore, in step S3, the four influencing factors and independent variables are ultrasonic power (X1: 252-360 W), ultrasonic temperature (X2: 30-50 °C), SDS concentration (X3: 10-60 mg / mL) and material-liquid ratio (X4: 1:10-1:20 g / mL).

[0047] In this embodiment, Design Expert 11 software was used to fit a second-order polynomial equation to the experimental data in order to predict the optimal process conditions and quantify the interactions between the various factors. The quadratic equation for the effects of the complete independent variables on the extraction yields of ISO, ISQ, K3R, and AST is as follows:

[0048]

[0049] Where Y is the predicted response, X i and X j are independent variables, β0, β i , β ij , β ii are the coefficients in the intercept term, linear term, quadratic term, and interaction term, respectively. The response surface quadratic model was verified using analysis of variance (ANOVA), and the significance of each item was tested. The design of the influencing factor level is shown in Table 1 below:

[0050] Table 1 Factors and levels used in response surface analysis

[0051] Table 3.3 Factors and levels used in BBD experimental.

[0052]

[0053] Furthermore, in step S4, the deep eutectic solvent is one of a choline lactic acid-water system or supercritical CO2.

[0054] Furthermore, in step S5, the molecular dynamics simulation software is GROMACS.

[0055] Furthermore, in step S5, the specific analysis content of the FT-IR analysis is to compare the changes in characteristic peaks of the cell wall components of Trifoliate Green before and after extraction, so as to reveal the damage mechanism of the surfactant on the cell wall.

[0056] Furthermore, in step S5, the SEM / TEM observation involves observing the morphological changes of plant cells before and after ultrasonic treatment using a scanning or transmission electron microscope to clarify the effects of the cavitation effect and the synergistic effect of the surfactant on the permeability of the cell wall.

[0057] Furthermore, in step S5, the Fick's second law model is specifically used to calculate the diffusion coefficient of flavonoid glycosides, and to illustrate the mechanism by which surfactants enhance the mass transfer rate.

[0058] In the present invention, six main factors affecting the extraction rates of ISO, ISQ, K3R and AST in the root tubers of Tripterygium wilfordii were tested individually, as follows:

[0059] (1) Screening of surfactant concentration: SDS was selected as the optimal surfactant based on the experimental results. SDS was accurately weighed and prepared with ultrapure water at concentrations of 10, 20, 35, 50, 65, and 80 mg / mL, respectively. Six portions of 1 g of Tripterygium wilfordii powder were weighed and 10 mL of surfactant aqueous solution was added to each portion at a solid-liquid ratio of 1:10 (g / mL). After soaking overnight, the mixture was ultrasonically treated at 360 W power and 30°C for 20 min. The extract was centrifuged at 10,000 rpm for 10 min, and the supernatant was filtered through a 0.22 μm filter membrane. The contents of ISO, ISQ, K3R, and AST were simultaneously detected by HPLC, and the extraction rate under each concentration condition was calculated.

[0060] Furthermore, since organic solvents are usually used to extract the active ingredients of flavonoids in Tripterygium wilfordii, it is particularly important to select a safe, green, efficient and low-cost extraction solvent to replace the traditional organic solvents with high toxicity and high cost in the extraction process. Figure 2 It can be seen that by comprehensively comparing the extraction rate and saturated solubility of various surfactants, sodium dodecyl sulfate (SDS) was finally determined as the optimal extraction solvent. The saturated solubility of SDS for the four flavonoid glycosides is higher than that of distilled water and other surfactants, which is beneficial to the extraction of flavonoid glycosides. In terms of economic factors, the price of SDS is much lower than other solvents. According to the physicochemical properties of surfactants, when the concentration of surfactant monomers reaches the critical micelle concentration (CMC), they will self-assemble to form micelle aggregates. This microstructural transformation significantly enhances the solubilization effect and provides theoretical support for the efficient extraction of flavonoid glycosides. Therefore, this study selected SDS as the extraction solvent

[0061] (2) Screening of material-liquid ratio: The concentration of SDS aqueous solution is 35 mg / mL. Weigh 1 g of Tripterygium wilfordii powder and set the material-liquid ratio to 1:10, 1:15, 1:20, 1:25, 1:30, and 1:35 (g / mL). Add surfactant aqueous solution and mix well. Soak overnight and extract for 20 min at 30°C with an ultrasonic power of 360 W. The extracts were treated in the same way and then analyzed by HPLC to investigate the effect of different material-liquid ratios on the extraction efficiency.

[0062] Furthermore, the selection of the material-liquid ratio is also important for the extraction of target active ingredients from the root tubers of Tripterygium wilfordii. Increasing the solvent volume can significantly improve the dispersion state of the material, expand the solid-liquid contact interface, and promote the diffusion and mass transfer of flavonoids to the solvent phase. Figure 3As shown, the extraction yield peaked at a solid-liquid ratio of 1:15 (g / mL). However, increasing the ratio further may limit the dissolution of the active ingredient, resulting in peak dissolution efficiency of the target ingredient, increased solvent consumption and cost, and the dissolution of other impurities, further compromising the extraction effect. Due to prolonged ultrasonic treatment, the transient high temperature and pressure generated, along with the released free radicals, led to the degradation of flavonoid glycosides. Therefore, a solid-liquid ratio of 1:15 (g / mL) was selected for the next step of the experiment.

[0063] (3) Screening of ultrasonic time: Under the conditions of 35 mg / mL SDS solution and 1:15 (g / mL) solid-liquid ratio, the extraction time was set to 2, 10, 15, 20, 25, and 35 min, respectively. Each sample was ultrasonically treated at 360W and 30°C. The extract was standardized and then detected by HPLC.

[0064] Furthermore, the extraction efficiency of active ingredients, compound stability, and extraction cost are closely related to ultrasonic time. The extraction rate of flavonoid glycosides increases with the increase of ultrasonic extraction time. This may be because the solvent dissolves surface compounds and intracellular components through diffusion and penetration processes respectively during this stage. Figure 4 It can be seen that the cavitation effect and microbubble dynamic behavior induced by ultrasound can effectively destroy the cell wall structure and reduce the mass transfer resistance in the initial stage (0-15min). However, continuous ultrasound treatment will lead to local extreme conditions (high temperature, high pressure, and free radical generation), triggering degradation reactions such as flavonoid glycoside bond breakage, resulting in a decrease in the extraction rate. Therefore, the next step of research was carried out with a single ultrasound of 15min as the response surface optimization condition.

[0065] (4) Screening of ultrasonic times: Under the conditions of a fixed material-liquid ratio of 1:15 (g / mL), 360W, 30℃, and an SDS concentration of 35 mg / mL, the number of extractions was set to 1, 2, 3, 4, 5, and 6 times, respectively. Each extraction lasted 20 minutes. The supernatant was collected by centrifugation at 10,000 rpm for 10 minutes, filtered through a 0.22 μm filter membrane, and quantitatively analyzed by HPLC to determine the optimal number of ultrasonic times.

[0066] Further, if Figure 5 The results showed that the dissolution of the target component was limited during a single ultrasonic treatment. As the number of extractions increased, the extraction rate gradually increased. When the extraction was repeated three times, the extraction rate of the target component was the highest. However, excessive ultrasonic extraction destroyed the molecular structure of the active ingredient, resulting in a decrease in the extraction rate. At the same time, excessive solvent consumption caused waste, increased operating costs, and did not meet the requirements of a green process. Comprehensively selecting two ultrasonic extractions as the optimal process parameters.

[0067] (5) Screening of ultrasonic power: Under the conditions of a solid-liquid ratio of 1:15 (g / mL) and an SDS concentration of 35 mg / mL, ultrasonic treatments were performed twice (20 min each). Ultrasonic extractions were performed at 95 W, 162 W, 198 W, 252 W, 306 W, and 360 W, respectively, and ultrasonic extraction was performed twice at 30°C. The extracts were centrifuged at 10,000 rpm for 10 min, and the supernatant was collected and filtered through a 0.22 μm filter membrane. The effects of different powers on the extraction yields of ISO, ISQ, K3R, and AST were analyzed by HPLC.

[0068] Further, by Figure 6 The extraction rates of ISO, ISQ, K3R, and AST increased with increasing ultrasonic power. When the ultrasonic power reached 306 W, cavitation intensity and mechanical vibration reached an optimal synergistic state, maximizing cell wall rupture efficiency. However, continued high-power ultrasonic extraction produced ineffective bubbles, which triggered cavitation saturation and disrupted the flavonoid glycoside structure, reducing the yield of the target active ingredients. Therefore, 306 W was selected as the condition for further BBD experiments.

[0069] (6) Screening of ultrasonic temperature: 6 portions of 1g of Tripterygium wilfordii powder were accurately weighed, and a 35mg / mL SDS aqueous solution was added at a solid-liquid ratio of 1:15 (g / mL) and mixed evenly. The mixture was soaked overnight and ultrasonically extracted at 360W. The ultrasonic temperatures were set to 25℃, 30℃, 40℃, 60℃, 70℃, and 80℃, respectively. Ultrasonic extraction was repeated twice, each time for 20 minutes. After the extraction, the mixture was centrifuged at 10,000rpm for 10 minutes, the supernatant was collected, filtered through a 0.22μm filter membrane, and analyzed by HPLC to detect ISO, ISQ, K3R, and AST in the Tripterygium wilfordii root tubers.

[0070] Furthermore, when the temperature rises to 40 °C, the solvent viscosity and surface tension decrease simultaneously, thereby improving the cavitation efficiency, and the increase in molecular diffusion rate promotes the dissolution of flavonoid glycosides, such as Figure 7 As shown, as the temperature exceeds 40°C, some flavonoid glycosides undergo oxidative decomposition, destabilizing the hydrogen bonds between hydrogen donors and hydrogen acceptors. Thermal effects accelerate the oxidative decomposition of flavonoid glycosides, offsetting the increased mass transfer rate caused by high temperatures. Therefore, 40°C was selected as the experimental condition for response surface optimization.

[0071] Furthermore, the coefficient of variation (CV, standard deviation: mean) is used to examine the degree of dispersion of each factor. Figure 8 The influence of the six factors was ranked as follows: ultrasonic power > ultrasonic temperature > surfactant concentration > solid-liquid ratio > ultrasonic time > ultrasonic times. Therefore, the top four factors with the greatest impact on the experimental results (ultrasonic power, ultrasonic temperature, surfactant concentration, and solid-liquid ratio) were selected as the main factors affecting the extraction rate for subsequent response surface experiments.

[0072] In the present invention, the experimental results of optimization of the process parameters of ultrasound-assisted micellar medium extraction are as follows: based on single-factor experiments and response surface optimization method, six factors affecting the extraction rates of ISO, ISQ, K3R and AST are selected. By comparing the coefficient of variation (CV), the four factors with the greatest influence on the target components are screened as the main factors of the response surface optimization experiment, namely ultrasonic power (X1), ultrasonic temperature (X2), surfactant concentration (X3), and material-liquid ratio (X4). Each group of experiments is repeated 3 times, among which YISO, YISQ, YK3R, and YAST are the dependent variables of ISO, ISQ, K3R, and AST extraction rates, respectively, as shown in Table 2 below:

[0073] Table 2 BBD experimental results

[0074]

[0075]

[0076] In the present invention, response surface experimental design and result analysis were performed: according to the BBD experiment, the ultrasonic power, ultrasonic temperature, SDS concentration and material-liquid ratio were optimized respectively, and the extraction rates of ISO, ISQ, K3R and AST were used as response values respectively. The 29 groups of experimental data shown in Tables 3-6 were processed by variance analysis and a quadratic polynomial regression model was constructed for fitting test, which finally confirmed the significance level of the model.

[0077] Table 3 Variance analysis of the extraction rate of isoorientin

[0078]

[0079] Table 4 Variance analysis of isoquercetin extraction rate

[0080]

[0081]

[0082] Table 5 Variance analysis of kaempferol-3-O-rutinoside extraction rate

[0083]

[0084] Table 6 Variance analysis of astragalin extraction rate

[0085]

[0086] In the present invention, ISO, ISQ, K3R, AST extraction rate model fitting: based on analysis of variance (ANOVA), quadratic model is verified (table 7), the F values of ISO, ISQ, K3R and AST are respectively 7.93, 4.51, 8.67 and 22.07, and the P values are respectively 0.2717, 0.153, 0.1899, 0.22589, all greater than 0.05, showing that the lack-of-fit term of model is not remarkable. In signal-to-noise ratio (S / N) index, 8.232 (ISO), 6.6389 (ISQ), 8.5517 (K3R) and 16.0237 (AST) are all greater than 4, indicating that the ratio signal is sufficient, and model is applicable to experimental analysis. The relationship between independent variable and response variable can be reflected by the quadratic polynomial equation that response surface methodology obtains, and the regression equation of ISO, ISQ, K3R, AST is as follows:

[0087] Table 7 Regression equations of isoorientin, isoquercetin, kaempferol-3-O-rutinoside, and astragaloside

[0088]

[0089] In the present invention, the effects of different interactive factors on the extraction rate: in order to explore the effects of ultrasonic power, ultrasonic temperature, SDS concentration, solid-liquid ratio and the interaction of other two factors on the target active ingredients.

[0090] When the extraction rate of ISO is the vertical axis and the above two factors are the horizontal axis, the response surface diagram is drawn as follows Figure 9 As shown, the upper part of the figure is A, B, and C from left to right, where A is the effect of the interaction between ultrasonic temperature and ultrasonic power on the ISO extraction rate, B is the effect of the interaction between SDS concentration and ultrasonic power on the ISO extraction rate, and C is the effect of ultrasonic power and solid-liquid ratio on the ISO extraction rate; the lower part of the figure is D, E, and F from left to right, where D is the effect of ultrasonic temperature and SDS concentration on the ISO extraction rate, E is the effect of solid-liquid ratio and ultrasonic temperature on the ISO extraction rate, and F is the effect of SDS concentration and solid-liquid ratio on the ISO extraction rate.

[0091] Further, if Figure 9 As shown in Figure A, when the solid-liquid ratio is 1:15 (g / mL) and the ultrasonic time is fixed at 15 minutes, the extraction rate of ISO increases continuously with increasing extraction power. After exceeding a certain threshold, the extraction rate gradually decreases. This may be because the continuous increase in ultrasonic power generates a large number of bubbles, triggering cavitation saturation, which leads to the destruction of the structure of the target active ingredient and the decrease in extraction rate.

[0092] Furthermore, when the ultrasonic temperature was 40 °C and the solid-liquid ratio was 1:15 (g / mL) as fixed conditions, the interactive effects of SDS concentration and ultrasonic power on ISO extraction efficiency were as follows: Figure 9 As shown in Figure B, when sonication time remains constant, the ISO extraction yield increases with increasing SDS concentration, then gradually decreases. While the ISO extraction yield remains proportional to the extraction time at a fixed SDS concentration, increasing the SDS concentration leads to a decrease in the ISO extraction yield. This is likely due to the implosion and cavitation of microbubbles, which rupture the cell walls and damage the plant cell structure. Prolonged sonication can lead to degradation of the target active ingredient, resulting in a decrease in the ISO extraction yield.

[0093] Furthermore, when the ultrasonic time was fixed at 15 min and the ultrasonic temperature at 40 °C, the interactive effects of the solid-liquid ratio and ultrasonic power on the ISO extraction rate were as follows: Figure 9 As shown in Figure B. Under the condition of constant ultrasonic power, the extraction rate of ISO increased significantly with the increase of the material-liquid ratio. However, as the material-liquid ratio continued to rise, the solubility of the material in the solution gradually reached a saturated state, causing the ISO extraction rate to gradually decrease and approach equilibrium after reaching a certain threshold.

[0094] Furthermore, when the solid-liquid ratio and ultrasonic power are fixed, the interactive effects of ultrasonic temperature and SDS concentration on ISO extraction efficiency are as follows: Figure 9 Figure A shows the extraction efficiency of ISO when the ultrasonic temperature remains constant. The extraction efficiency of ISO increases with increasing SDS concentration, initially increasing but then gradually decreasing. This is likely due to the increasing viscosity of the solvent system, which prevents the SDS solution from fully penetrating the cells, thus affecting the extraction efficiency.

[0095] Further, if Figure 9 As shown in Figure A, when the ultrasonic time and SDS concentration conditions remain unchanged and the ultrasonic temperature is constant, the extraction rate of ISO first increases and then gradually decreases with the change of the solid-liquid ratio. When it exceeds 1:15 (g / mL), the extraction rate gradually decreases.

[0096] Furthermore, when the ultrasonic temperature was maintained at 40 °C and the ultrasonic power was 306 W, the interactive effects of the solid-liquid ratio and SDS concentration on the ISO extraction rate were as follows: Figure 9 As shown in Figure A, at a fixed solid-liquid ratio, the ISO extraction rate initially increases and then decreases with increasing SDS concentration. At a fixed SDS concentration, higher solid-liquid ratios lead to higher extraction efficiency. However, above a certain threshold, the ISO extraction rate decreases and then stabilizes. Continuously increasing the solid-liquid ratio not only wastes resources but also runs counter to the sustainable development philosophy of efficient energy conservation and emission reduction.

[0097] When the extraction rate of ISQ is taken as the vertical axis and the above two factors are taken as the horizontal axis, the response surface graph is drawn as follows: Figure 10As shown, the upper part of the figure is A, B, and C from left to right, where A is the effect of the interaction between ultrasonic temperature and ultrasonic power on the ISQ extraction rate, B is the effect of the interaction between SDS concentration and ultrasonic power on the ISQ extraction rate, and C is the effect of ultrasonic power and solid-liquid ratio on the ISQ extraction rate; the lower part of the figure is D, E, and F from left to right, where D is the effect of ultrasonic temperature and SDS concentration on the ISQ extraction rate, E is the effect of solid-liquid ratio and ultrasonic temperature on the ISO extraction rate, and F is the effect of SDS concentration and solid-liquid ratio on the ISQ extraction rate.

[0098] Furthermore, under the conditions of a fixed solid-liquid ratio of 1:15 (g / mL) and ultrasonic time of 15 min, the interactive effect of ultrasonic temperature and power on the ISQ extraction rate was as follows: Figure 10 As shown in Figure A, when the ultrasonic temperature is fixed, the extraction rate first increases and then decreases with increasing ultrasonic power. This phenomenon may be attributed to the supersaturation of the cavitation effect caused by excessive ultrasonic power, which damages the molecular structure of the active ingredient and reduces the extraction rate.

[0099] Furthermore, when the ultrasonic temperature was kept constant at 40 °C and the material-liquid ratio was maintained at 1:15 (g / mL), the interaction between SDS concentration and ultrasonic power on the extraction rate was as follows: Figure 10 As shown in Figure B, at a given sonication duration, the ISQ extraction yield exhibited a parabolic change with increasing SDS concentration, while increasing sonication time was positively correlated with the extraction yield. However, excessive SDS bioconcentration caused a decrease in the extraction yield. This mechanism may be related to excessive cell wall lysis caused by cavitation-induced microbubble implosion, while excessive sonication can accelerate the oxidative degradation of the target active ingredients.

[0100] Furthermore, under the fixed combination of ultrasonic time of 15 min and temperature of 40 °C, the interactive effect of solid-liquid ratio and ultrasonic power on ISQ extraction rate is as follows: Figure 10 As shown in Figure B, when the extraction power remains constant, the ISQ extraction rate increases with the material-liquid ratio, reaching a critical point and then gradually stabilizing. This may be because the solute solubility reaches saturation at a certain material-liquid ratio. Further increasing the solvent ratio no longer improves mass transfer efficiency, but instead reduces the ISQ extraction rate due to system dilution.

[0101] Furthermore, when the material-liquid ratio was kept at 1:15 (g / mL) and the power was 306W, the interaction between temperature and SDS concentration was as follows: Figure 10 Figure 1 (A). Under constant temperature, the ISQ extraction rate initially increases and then decreases with increasing SDS concentration, with the inflection point associated with a sudden change in the solvent system's viscosity. The increased viscosity caused by high SDS concentrations hinders the osmotic diffusion of the solvent into the cells, thus creating mass transfer resistance.

[0102] Furthermore, under the fixed conditions of ultrasonic time of 15 min and SDS concentration of 35 mg / mL, the interaction between ultrasonic temperature and material-liquid ratio is as follows: Figure 10 As shown in Figure A, when the ultrasonic temperature remains constant, the extraction yield increases with increasing material-to-liquid ratio. However, after exceeding the critical value of 1:15, the extraction yield exhibits a negative growth trend. This trend confirms the solvent-solute equilibrium theory: excess solvent dilutes the concentration of the active ingredient per unit volume, leading to a decrease in the ISQ extraction yield.

[0103] Furthermore, when the temperature is constant at 40 °C and the ultrasonic power is 306 W, the interaction between the material-liquid ratio and the SDS concentration is as follows: Figure 10 As shown in Figure A, under a fixed solid-liquid ratio, the ISQ extraction yield initially increases and then decreases with increasing SDS concentration. When the SDS concentration is fixed, the extraction yield and solid-liquid ratio are positively correlated up to a threshold value, then level off. However, excessive pursuit of a high solid-liquid ratio results in solvent waste, violating the principles of green chemistry. A balance must be struck between extraction yield and resource consumption.

[0104] When the AST extraction rate is the vertical axis and the above two factors are the horizontal axis, the response surface graph is drawn as follows Figure 11 As shown, the upper part of the figure is A, B, and C from left to right, where A is the effect of the interaction between ultrasonic temperature and ultrasonic power on the AST extraction rate, B is the effect of the interaction between SDS concentration and ultrasonic power on the AST extraction rate, and C is the effect of solid-liquid ratio and ultrasonic power on the AST extraction rate; the lower part of the figure is D, E, and F from left to right, where D is the effect of ultrasonic temperature and SDS concentration on the AST extraction rate, E is the effect of ultrasonic temperature and solid-liquid ratio on the AST extraction rate, and F is the effect of SDS concentration and solid-liquid ratio on the AST extraction rate.

[0105] Furthermore, Figure 11 Figure A shows the interactive effect of ultrasonic temperature and power on the extraction yield of K3R under the fixed conditions of a solid-liquid ratio of 1:15 (g / mL) and an ultrasonic time of 15 minutes. When the temperature is fixed, the extraction yield increases with the gradient of ultrasonic power, rising rapidly initially, then declining after reaching a peak. Excessive ultrasonic power causes cavitation bubble collapse, resulting in a stronger sonochemical effect and thus degradation of the active ingredient.

[0106] Further, by Figure 11Figure B shows the interaction between SDS concentration and ultrasonic power on the extraction yield under the conditions of a fixed ultrasonic temperature of 40°C and a solid-to-liquid ratio of 1:15 (g / mL). Under fixed ultrasonic conditions, the extraction yield reached its highest at an SDS concentration of 35 mg / mL. However, the extraction yield gradually decreased with increasing concentration. This may be due to the increased implosion intensity of cavitation microbubbles with power, leading to excessive lysis of plant tissue cell walls. Furthermore, prolonged ultrasonic treatment induced oxidative degradation of heat-sensitive components, resulting in a decrease in the extraction yield of K3R.

[0107] Furthermore, Figure 11 Figure B shows the interactive effect of ultrasonic power and liquid-to-liquid ratio on the extraction efficiency of K3R under the fixed conditions of 15 minutes of ultrasonication and 40°C of temperature. At a fixed ultrasonic power, the extraction efficiency of K3R decreases significantly with increasing liquid-to-liquid ratio, reaching the maximum extraction efficiency point before stabilizing. This phenomenon may be because once the solute-to-solvent ratio exceeds the critical value, the driving force for mass transfer no longer increases, and dilution of the solvent system actually reduces the concentration of the active ingredient per unit volume.

[0108] Furthermore, Figure 11 Figure A shows the interaction between ultrasonic temperature and SDS concentration on the extraction yield of K3R under the fixed conditions of a solid-liquid ratio of 1:15 (g / mL) and an ultrasonic power of 306 W. Under constant ultrasonic temperature, the extraction yield of K3R increases with increasing SDS concentration, with the turning point closely related to the sudden change in the rheological properties of the solvent system. High SDS concentrations increase the solution viscosity, creating a mass transfer boundary layer resistance that hinders the diffusion of solute molecules and reduces the effective bioactive components.

[0109] Furthermore, Figure 11 Figure A reveals the interaction between sonication temperature and the solid-liquid ratio under the fixed conditions of 15 minutes of sonication time and 35 mg / mL SDS concentration. At a fixed sonication temperature, excess solvent weakens the interactions between solute molecules, reducing the effective extraction concentration gradient. Consequently, the extraction yield of K3R increases with increasing solid-liquid ratio, but begins to decline after exceeding 1:15 (g / mL).

[0110] Furthermore, Figure 11 Figure A shows the interaction between the solid-liquid ratio and SDS concentration under fixed conditions of an ultrasonic temperature of 40°C and an ultrasonic power of 306W. Under these fixed solid-liquid ratios, the K3R extraction yield initially increases and then decreases with increasing SDS concentration. When the SDS concentration is constant, the K3R extraction yield is positively correlated with the solid-liquid ratio until it reaches a threshold, where it stabilizes. However, pursuing an excessive solid-liquid ratio not only increases solvent consumption but also potentially increases the cost of subsequent separation and purification, violating the principles of green extraction.

[0111] When the AST extraction rate is the vertical axis and the above two factors are the horizontal axis, the response surface graph is drawn as follows Figure 12 As shown, the upper part of the figure is A, B, and C from left to right, where A is the effect of the interaction between ultrasonic temperature and ultrasonic power on the K3R extraction rate, B is the effect of the interaction between SDS concentration and ultrasonic power on the K3R extraction rate, and C is the effect of ultrasonic power and solid-liquid ratio on the ISO extraction rate; the lower part of the figure is D, E, and F from left to right, where D is the effect of ultrasonic temperature and SDS concentration on the K3R extraction rate, E is the effect of solid-liquid ratio and ultrasonic temperature on the ISO extraction rate, and F is the effect of solid-liquid ratio and SDS concentration on the K3R extraction rate.

[0112] Furthermore, under the conditions of a fixed solid-liquid ratio of 1:15 (g / mL) and an ultrasonic time of 15 min, the synergistic effect of ultrasonic temperature and ultrasonic power on AST extraction efficiency was as follows: Figure 12 As shown in Figure A, experimental data show that when the temperature remains constant, the AST extraction rate initially increases and then decreases with increasing ultrasonic power. This is likely due to the excessive cavitation bubbles at power levels exceeding 306 W, which trigger a "cavitation shielding" effect that in turn destroys the structure of the target active ingredient, reducing the AST extraction rate.

[0113] Furthermore, when the ultrasonic temperature was fixed at 40 °C and the material-liquid ratio was 1:15 (g / mL), the interaction between SDS concentration and ultrasonic power was as follows: Figure 12 As shown in Figure B. When the SDS concentration increased from 20 mg / mL to 35 mg / mL, the extraction rate increased significantly. However, when the concentration exceeded 40 mg / mL, the solution viscosity increased, hindering effective penetration and resulting in a decrease in the AST extraction rate.

[0114] Furthermore, under the conditions of fixed time of 15 min and ultrasonic temperature of 40 °C, the interaction between material-liquid ratio and power is as follows: Figure 12 As shown in Figure B, when the solid-liquid ratio increased from 1:10 to 1:15 (g / mL), the solvent fully infiltrated the solution, resulting in an improved extraction rate. However, when the solid-liquid ratio exceeded 1:15, the diffusion force weakened, causing the AST extraction rate to decrease and then level off.

[0115] Furthermore, when the material-liquid ratio was fixed at 1:15 (g / mL) and the ultrasonic power was 306 W, the synergistic effect of ultrasonic temperature and SDS concentration was as follows: Figure 12 As shown in Figure A, when the SDS concentration increased from 25 mg / mL to 35 mg / mL, the extraction rate significantly improved due to the reduction in the solvent's surface tension. However, above 40 mg / mL, micelle formation entrapped the active ingredient, resulting in a decrease in the AST extraction rate.

[0116] Furthermore, under the conditions of fixed ultrasonic time of 15 min and SDS concentration of 35 mg / mL, the interaction between ultrasonic temperature and material-liquid ratio is as follows: Figure 12 As shown in Figure A. When the solid-liquid ratio is 1:15 (g / mL), the AST extraction rate reaches its peak. When the ratio exceeds this value, the excess solvent leads to a decrease in mass transfer efficiency. At the same time, temperatures exceeding 50°C will cause thermal denaturation of the target component, resulting in a decrease in the AST extraction rate.

[0117] Furthermore, when the ultrasonic temperature is fixed at 40 °C and the ultrasonic power is fixed at 306 W, the interaction between the material-liquid ratio and the SDS concentration is as follows: Figure 12 As shown in Figure A, extraction efficiency is highest when the material-liquid ratio is 1:15 (g / mL) and the SDS concentration is 35 mg / mL. However, a material-liquid ratio exceeding 1:20 results in solvent waste, reducing utilization, and SDS concentrations exceeding 40 mg / mL form a micelle barrier, reducing extraction efficiency. This trend demonstrates the importance of parameter optimization for improving resource efficiency.

[0118] In the present invention, statistical software analysis was performed using a combination of model fitting and regression equations to determine the optimal process conditions for ultrasound-assisted micellar medium extraction of Tripterygium wilfordii: ultrasonic power: 100W; ultrasonic temperature: 50°C; SDS concentration: 35mg / mL; and solid-liquid ratio: 1:15 (g / mL). The predicted values for ISO, ISQ, K3R, and AST were 6.704μg / g, 17.471μg / g, 18.242μg / g, and 16.033μg / g, respectively. Three parallel experiments were conducted under these conditions to verify the accuracy of the model. The extraction rates for ISO, ISQ, K3R, and AST were 5.856μg / g, 15.296μg / g, 17.474μg / g, and 15.683μg / g, respectively. The experimental extraction rates were in good agreement with the predicted values, indicating that the extraction method is stable and highly reproducible.

[0119] In the present invention, the process of this design is green and environmentally friendly, and sodium lauryl sulfate is used as a natural surfactant to replace traditional organic solvents (such as methanol).

[0120] High efficiency and energy saving: Ultrasonic assistance shortens extraction time and reduces energy consumption. High selectivity: Micellar medium reduces impurity dissolution.

[0121] In the present invention, the process parameters for extracting isoorientin, isoquercetin, kaempferol-3-O-rutinoside and astragaloside from the tuberous roots of Tripterygium wilfordii using an ultrasound-assisted micellar medium of sodium dodecyl sulfate aqueous solution were optimized, and the effects of the type and concentration of surfactant, material-liquid ratio, ultrasonic power, ultrasonic time, ultrasonic times and ultrasonic temperature on the target components were investigated. The optimal extraction process parameters were determined as follows: sodium dodecyl sulfate (SDS) concentration: 35 mg / mL, ultrasonic power: 100 W, and ultrasonic temperature: 50°C. Under the above optimal extraction process conditions, the average extraction rates of isoorientin, isoquercetin, kaempferol-3-O-rutinoside and astragaloside were 5.856 μg / g, 15.296 μg / g, 17.474 μg / g and 15.683 μg / g, respectively. The differences in the extraction of various components are related to the polar micelle solubilization ability of flavonoid glycosides. This process replaces traditional organic solvents with surfactants, providing a feasible solution for the green extraction of the active ingredients of Tripterygium wilfordii. The feasibility of industrial scale-up can be further explored in the future.

[0122] The above disclosure is only a preferred embodiment of the present invention, and certainly cannot be used to limit the scope of the rights of the present invention. Ordinary technicians in this field can understand that all or part of the processes of the above embodiment and equivalent changes made in accordance with the claims of the present invention are still within the scope of the invention.

Claims

1. A process optimization method for green extraction of flavonoid glycosides from Tripterygium wilfordii using ultrasound-assisted surfactants, characterized in that: The steps include: Step S1: Accurately weigh 1 g of Tripterygium wilfordii powder and mix it evenly with 5% aqueous solutions of 9 surfactants at a solid-liquid ratio of 1:10 (g / mL). Extraction was performed under the same ultrasonic conditions. Three parallel experiments were conducted in each group to investigate the saturated solubility of ISO, ISQ, K3R, and AST in Tripterygium wilfordii roots with different surfactant aqueous solutions and the extraction rate of target components in Tripterygium wilfordii roots by ultrasonic-assisted extraction, so as to determine the optimal surfactant type. Step S2: Screening out 6 main factors that affect the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii, conducting experiments on individual factors, and conducting 3 parallel experiments in each group, and then screening out the final factor that affects the extraction rates of ISO, ISQ, K3R, and AST in the tuberous roots of Tripterygium wilfordii; Step S3: Box-Behnken Design was used to further optimize the extraction process conditions of ISO, ISQ, K3R, and AST from the root tubers of Tripterygium wilfordii using ultrasound-assisted micellar medium, and four influencing factors were selected as independent variables; Step S4: performing auxiliary extraction in combination with a deep eutectic solvent, recording and comparing the extraction effects under different solvent systems, so as to minimize the amount of organic solvent used and achieve friendly extraction; Step S5: Molecular dynamics simulation software is used to simulate the interaction process between the optimal surfactant type and flavonoid glycosides in Tripterygium wilfordii, and the variables of the final factor and the four influencing factors are controlled. At the same time, FT-IR analysis, SEM / TEM observation and Fick's second law model are combined to comprehensively explain the synergistic mechanism of surfactants on the transmembrane transport of flavonoid glycosides, and finally complete the optimization of the Tripterygium wilfordii flavonoid glycoside process.

2. The process optimization method for green extraction of flavonoid glycosides of trifoliate green by ultrasound-assisted surfactant as claimed in claim 1, characterized in that: In step S1, the nine surfactants are sodium lauryl sulfate, sodium lauryl sulfonate, sodium lauryl sulfate, cocamidopropyl betaine, dodecyl dimethyl betaine, hexadecyl trimethyl ammonium chloride, alkyl glucoside, sucrose fatty acid ester and polyvinyl alcohol, and the extraction power is set to 360 W, the ultrasonic temperature is below 30° C., and the extraction time is 20 min.

3. The process optimization method for green extraction of flavonoid glycosides of trifoliate green by ultrasound-assisted surfactant as claimed in claim 2, characterized in that: In step S2, the six main factors affecting the extraction rates of ISO, ISQ, K3R and AST in the root tubers of Tripterygium wilfordii are ultrasonic power, solid-liquid ratio, ultrasonic times, ultrasonic time, surfactant concentration and ultrasonic temperature.

4. The process optimization method for green extraction of flavonoid glycosides of trifoliate green by ultrasound-assisted surfactant as claimed in claim 3, characterized in that: In step S3, the four influencing factors and independent variables are ultrasonic power (X1: 252-360 W), ultrasonic temperature (X2: 30-50 °C), SDS concentration (X3: 10-60 mg / mL) and solid-liquid ratio (X4: 1:10-1:20 g / mL).

5. The process optimization method for green extraction of flavonoid glycosides of trifoliate green by ultrasound-assisted surfactant as claimed in claim 4, characterized in that: In step S4, the deep eutectic solvent is one of a choline lactic acid-water system or supercritical CO2.

6. The process optimization method for green extraction of flavonoid glycosides of trifoliate green by ultrasound-assisted surfactant as claimed in claim 5, characterized in that: In step S5, the molecular dynamics simulation software is GROMACS.

7. The process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactant as claimed in claim 6, characterized in that: In step S5, the specific analysis content of the FT-IR analysis is to compare the changes in characteristic peaks of the cell wall components of Trifoliate Green before and after extraction, and to reveal the damage mechanism of the surfactant on the cell wall.

8. The process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactant as claimed in claim 7, characterized in that: In step S5, the SEM / TEM observation is to observe the morphological changes of plant cells before and after ultrasonic treatment using a scanning or transmission electron microscope to clarify the effects of the cavitation effect and the synergistic effect of the surfactant on the permeability of the cell wall.

9. The process optimization method for green extraction of trifoliate green flavonoid glycosides by ultrasound-assisted surfactant as claimed in claim 8, characterized in that: In step S5, the Fick's second law model is specifically used to calculate the diffusion coefficient of flavonoid glycosides, and to illustrate the mechanism by which surfactants enhance the mass transfer rate.

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