Glehnia root active ingredient extract as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of traditional Chinese medicine extraction and application, and particularly relates to a radix glehniae effective component extract and a preparation method and application thereof.The preparation method comprises the following steps of S1, pretreatment, S2, ultrasonic-assisted treatment, specifically, pretreated radix glehniae powder is extracted under the effect of ultrasonic waves, and an ultrasonic-assisted treatment solution is obtained; s3, performing enzymolysis treatment: adding cellulase, papain and saccharifying enzyme into the ultrasonic-assisted treatment solution, adjusting the pH value of the system to 5-6, and performing enzymolysis to obtain an enzymolysis extracting solution; s4, membrane separation: filtering the enzymolysis extracting solution to obtain a concentrated solution containing the effective component extract of the radix glehniae; s5, drying to obtain the radix glehniae effective component extract. According to the preparation method provided by the invention, the effective components in the radix glehniae can be efficiently extracted, the effective components are prevented from being damaged, and meanwhile, the effective components can be accurately applied to the field of lung health.

Description

Technical Field

[0001] This invention specifically relates to an extract of the effective components of North American ginseng, its preparation method and application, belonging to the field of traditional Chinese medicine extraction and application technology. Background Technology

[0002] With environmental changes, lifestyle shifts, and an aging population, lung health issues are becoming increasingly prominent. For example, lung damage caused by fine particulate matter (PM2.5) pollution is leading to an increasing incidence of lung inflammation, respiratory dysfunction, asthma, emphysema, and chronic bronchitis. Serious lung diseases such as lung cancer also pose a significant threat to human life and health.

[0003] In the prevention and treatment of lung diseases, traditional Chinese medicine (TCM) demonstrates irreplaceable advantages due to its unique theoretical system and characteristics. North American ginseng (Beisha ginseng) is a commonly used TCM herb. It is slightly cold in nature and has a sweet and slightly bitter taste. In terms of efficacy, it can clear lung heat, nourish lung yin, and replenish stomach fluids, thus promoting the production of body fluids. North American ginseng contains various components such as polysaccharides, coumarins, polyacetylenes, lignans, terpenes, and flavonoids. Existing research shows that North American ginseng can increase the ratio of human T cells, improve lymphocyte transformation rate, increase white blood cell count, enhance macrophage function, and improve B cell immune function, thereby strengthening the body's resistance, reducing disease, and even preventing cancer to some extent. However, traditional methods of using North American ginseng, such as decoction and boiling to extract active ingredients, suffer from problems such as nutrient destruction and weakening of active ingredients due to high temperatures.

[0004] In summary, existing technologies have obvious shortcomings. Developing a highly efficient method for extracting the active ingredients from North American ginseng while ensuring that these ingredients are not destroyed, and enabling precise application of these active ingredients in the field of lung health, has become an urgent problem to be solved. Summary of the Invention

[0005] This invention addresses the shortcomings of existing technologies by providing an extract of active ingredients from North American ginseng, its preparation method, and its application. This preparation method can efficiently extract the active ingredients from North American ginseng while ensuring that the active ingredients are not destroyed. At the same time, the active ingredients can be precisely applied to the field of lung health.

[0006] The technical solution of this invention to solve the above-mentioned technical problems is as follows: A method for preparing an extract of effective components of North American ginseng, the preparation method comprising the following steps: S1. Pretreatment: Take North American ginseng, dry it in the shade, crush it, and sieve it to obtain North American ginseng powder for later use. S2, Ultrasonic-assisted treatment: Take the North American ginseng powder pretreated in step S1, add water, and extract it under ultrasonic treatment to obtain an ultrasonic-assisted treatment solution. S3. Enzymatic hydrolysis: Add cellulase, papain and saccharifying enzyme to the ultrasonic-assisted treatment solution obtained in step S2, adjust the pH of the system to 5-6 and carry out enzymatic hydrolysis to obtain the enzymatic hydrolysis extract. S4. Membrane separation: The enzymatic hydrolysis extract obtained in step S3 is filtered to obtain a concentrated extract containing the effective components of North American ginseng. S5. Drying: Spray dry the concentrate obtained in step S4 to obtain the extract of active ingredients of North American ginseng.

[0007] Furthermore, in step S1, the sample is passed through an 8-12 mesh sieve; preferably 10 mesh.

[0008] Furthermore, in step S2, the ratio of water to North American ginseng powder is (20~50) mL:1g; preferably 30 mL:1g.

[0009] Furthermore, in step S2, the ultrasonic power is 250W~350W, preferably 300W; the temperature is 50℃~70℃, preferably 60℃; and the extraction time is 0.5h~2h, preferably 1h.

[0010] Furthermore, in step S3, the pH is preferably 5.

[0011] Furthermore, in step S3, the enzymatic hydrolysis temperature is 40℃~70℃, preferably 60℃; the enzymatic hydrolysis time is 2.5h~3.5h, preferably 3h.

[0012] Furthermore, in step S3, the total mass of cellulase, papain, and saccharifying enzyme is 1.0-4.0% of the mass of North American ginseng powder taken in step S2, preferably 3.0%; wherein the mass ratio of cellulase, papain, and saccharifying enzyme is (2.5-5):1:1, preferably 3:1:1.

[0013] Furthermore, in step S4, the filtration is ultrafiltration, which is performed using an ultrafiltration membrane, specifically a polyphenylsulfone ultrafiltration membrane with a molecular weight cutoff of 3.0 kDa.

[0014] Furthermore, in step S5, the inlet air temperature of the spray dryer is 160℃~200℃, preferably 180℃; the fan frequency is 35Hz~50Hz, preferably 45Hz; the feed flow rate is 15mL / min~20mL / min, preferably 17mL / min; and the outlet air temperature is 60℃~90℃, preferably 80℃.

[0015] The present invention also discloses an extract of the effective components of North American ginseng, which is prepared according to the preparation method provided by the present invention.

[0016] The present invention also discloses an application of an extract of active ingredients from Glehnia littoralis, which is applied to the field of improving lung health, wherein the field of lung health is any one of treating lung inflammation, improving lung function, preventing lung diseases, and adjuvant treatment of lung diseases.

[0017] Furthermore, the lung inflammation is treated with North American Ginseng Capsules.

[0018] Furthermore, the North American ginseng capsules contain North American ginseng active ingredient extract powder, dextrin, sorbitol, and magnesium stearate.

[0019] The beneficial effects of this invention are: (1) The method for preparing the effective extract of Glehnia littoralis provided by the present invention involves pre-treating Glehnia littoralis and using a combination of ultrasonic-assisted, enzymatic hydrolysis, and membrane separation technologies. This significantly improves the extraction rate of polysaccharides from Glehnia littoralis. Finally, a specific drying process is used to ensure that the extracted effective components are not destroyed and are well preserved. The method provided by the present invention can ensure that the polysaccharide extraction rate in the final effective extract of Glehnia littoralis is above 20%, which is significantly higher than that of traditional extraction methods (below 20%). When the preparation process is precisely controlled with the parameters of Example 1, the polysaccharide extraction rate can reach as high as 38%.

[0020] (2) The extract of active ingredients of North American ginseng prepared by the present invention has good effects in the treatment of lung inflammation, improvement of lung function and adjuvant treatment of lung cancer, providing a new and effective prevention and treatment approach for the field of lung health.

[0021] (3) North American ginseng is a traditional Chinese medicinal material with relatively high safety. The purification method and application method of the present invention do not introduce harmful chemical substances, and the prepared product has few toxic side effects and can be taken for a long time.

[0022] (4) According to different application needs, the active ingredient extract of North American ginseng is prepared into a variety of dosage forms, which makes it convenient for patients and consumers to choose and use, thus improving the applicability and market competitiveness of the product. Attached Figure Description

[0023] Figure 1 The graph shows the changes in polysaccharide extraction rate between Example 1 and Examples 4-6; Figure 2 The graph shows the changes in polysaccharide extraction rate between Example 1 and Examples 7-9; Figure 3 The graph shows the changes in polysaccharide extraction rate between Example 1 and Examples 10-12; Figure 4 The graph shows the changes in polysaccharide extraction rate between Example 1 and Examples 13-15; Figure 5This is a graph showing the change in polysaccharide extraction rate in Example 16; Figure 6 This is a graph showing the anti-inflammatory activity of the extract of active ingredients from *Adenophora stricta* in Example 1. Detailed Implementation

[0024] The specific embodiments of the present invention will be described in detail below. The present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used is for describing particular embodiments only and is not intended to limit the invention.

[0026] The North American ginseng used in this embodiment of the invention is derived from Laiyang Laihu ginseng, characterized by: slender cylindrical roots, 30-70cm long and 0.5-1.5cm in diameter. The surface is pale yellowish-white or yellowish-white, slightly rough, with fine longitudinal wrinkles and brownish-yellow dotted rootlet scars. It is brittle and easily broken; the cross-section shows a light yellowish-white bark and yellow xylem, with clear separation between bark and xylem.

[0027] A method for preparing an extract of effective components from *Adenophora stricta*, comprising the following steps: S1. Pretreatment: Take North American ginseng, dry it in the shade, crush it, and pass it through an 8-12 mesh sieve to obtain North American ginseng powder for later use. S2. Ultrasonic-assisted treatment: Take the pretreated North American ginseng powder from step S1, add water, and the ratio of water to North American ginseng powder is (20~50) mL: 1g. Extract under ultrasonic action to obtain an ultrasonic-assisted treatment solution. S3. Enzymatic hydrolysis: Add cellulase, papain and saccharifying enzyme to the ultrasonic-assisted treatment solution obtained in step S2, adjust the pH of the system to 5-6 and carry out enzymatic hydrolysis to obtain the enzymatic hydrolysis extract. S4. Membrane separation: The enzymatic hydrolysis extract obtained in step S3 is filtered to obtain a concentrated extract containing the effective components of North American ginseng. S5. Drying: Spray dry the concentrate obtained in step S4 to obtain the extract of active ingredients of North American ginseng.

[0028] Pretreatment is the foundation of the entire process. Air drying removes moisture to prevent component degradation, while pulverizing and sieving through an 8-12 mesh sieve increases the contact surface area for subsequent processing. A 10-mesh sieve is preferred, striking a balance between powder particle size and subsequent liquid extraction efficiency. This avoids excessively fine powder that would hinder filtration while ensuring sufficient specific surface area to promote the release of active ingredients. Pretreatment provides the optimal material foundation for the subsequent liquid-solid extraction process and is closely linked to subsequent steps.

[0029] Specifically, in step S2, the preferred ratio of water to North American ginseng powder is 30 mL: 1 g.

[0030] Specifically, in step S2, the ultrasonic power is 250W~350W, preferably 300W; the temperature is 50℃~70℃, preferably 60℃; and the extraction time is 0.5h~2h, preferably 1h.

[0031] The ultrasound-assisted treatment step adopted a specific liquid-to-solid ratio (the ratio of water to North American ginseng powder was (25~35) mL:1g). Through precise control of ultrasonic power and temperature parameters, the combination of these parameters not only played an individual role, but more importantly, created the optimal reaction substrate state for the subsequent enzymatic hydrolysis step. The cavitation effect, mechanical vibration and thermal effect of ultrasound worked together to create microcracks in the plant cell wall, which could effectively destroy the plant cell wall structure and accelerate the dissolution of the effective components in the cell into the aqueous phase. This step created favorable conditions for the subsequent enzymatic hydrolysis and significantly increased the contact area between the enzyme and the substrate.

[0032] In this embodiment of the invention, the water used in step S2 is purified water, but this does not constitute a limitation on the technology of the present invention. In practical applications, different types of purified water or drinking water can be selected according to the actual situation.

[0033] Specifically, in step S3, the pH is preferably 5.

[0034] Specifically, in step S3, the enzymatic hydrolysis temperature is 40℃~70℃, preferably 60℃; the enzymatic hydrolysis time is 1h~4h, preferably 3h.

[0035] Specifically, in step S3, the total mass of cellulase, papain and saccharifying enzyme is 1.0 to 4.0% of the mass of North American ginseng powder taken in step S2, preferably 3.0%; wherein the mass ratio of cellulase, papain and saccharifying enzyme is (2.5 to 5):1:1, preferably 3:1:1.

[0036] Cellulose, as a core component of the cell wall, plays a major barrier role in the dissolution of polysaccharides. Cellulase can effectively promote polysaccharide dissolution by hydrolyzing the cell wall, while preserving the original structure of intracellular substances. Papain is an endopeptidase containing thiol peptide chains, exhibiting both protease and esterase activity. In practice, it hydrolyzes proteins surrounding or intertwining with polysaccharides, opening the "channels" for polysaccharide dissolution and accelerating the entry of polysaccharides into the extract. Glucoamylase, also known as saccharifying enzyme, can increase the polysaccharide content in *Adenophora stricta* extract. Its core principle is the enzymatic transformation of insoluble and low-water-soluble carbohydrates in *Adenophora stricta*—by breaking specific glycosidic bonds, it converts carbohydrates (such as starch, oligosaccharides, and structural glycosides) that cannot be dissolved by conventional extraction processes into soluble polysaccharides or oligosaccharides, ultimately increasing the total amount of polysaccharides in the extract.

[0037] The enzymatic hydrolysis process of this invention employs a composite enzyme system composed of three enzymes in a specific ratio, which work synergistically under conditions of pH=5~6 and temperature of 55℃~65℃. The selection of this composite enzyme ratio and the precise control of the enzymatic hydrolysis conditions enable cellulase to primarily attack the cell wall structure, papain to remove protein impurities, and saccharifying enzyme to regulate the molecular weight of polysaccharides. The three work synergistically to thoroughly dismantle the plant tissue barrier, significantly improving the extraction efficiency and selectivity of the target components, and enhancing the purity and biological activity of polysaccharides.

[0038] Specifically, in step S4, the filtration is ultrafiltration, which is performed using an ultrafiltration membrane, specifically a polyphenylsulfone ultrafiltration membrane with a molecular weight cutoff of 3.0 kDa.

[0039] Membrane separation follows the enzymatic hydrolysis process, using ultrafiltration technology to filter the enzymatic extract. This process utilizes the precise sieving characteristics of ultrafiltration membranes to effectively retain large molecular impurities, such as unhydrolyzed polysaccharides and proteins, while allowing smaller molecules of the target active ingredients to pass through or remain in the concentrate, thus achieving the dual purpose of purification and concentration.

[0040] The preparation method of the active ingredient extract of Glehnia littoralis provided by this invention mainly removes macromolecular impurities with a molecular weight of 3.0 kDa through ultrafiltration in the membrane separation step.

[0041] Specifically, in step S5, the inlet air temperature of the spray dryer is 160℃~200℃, preferably 180℃; the fan frequency is 35Hz~50Hz, preferably 45Hz; the feed flow rate is 15mL / min~20mL / min, preferably 17mL / min; and the outlet air temperature is 60℃~90℃, preferably 80℃.

[0042] According to the preparation method of the active ingredient extract of *Adenophora stricta* provided by this invention, after obtaining a concentrated solution containing the active ingredient extract of *Adenophora stricta* through membrane separation, the final drying step adopts spray drying technology. The concentrated solution is converted into a dry powder under the preferred conditions of an inlet air temperature of 180°C, a fan frequency of 45Hz, a feed flow rate of 17mL / min, and an outlet air temperature of 80°C. This process, through strict control of hot air temperature and material flow rate, aims to rapidly evaporate moisture while avoiding excessive thermal stress that could lead to degradation or inactivation of heat-sensitive active ingredients, thereby obtaining a *Adenophora stricta* active ingredient extract with good flowability and high stability. The parameter settings of spray drying are interrelated with the physical properties of the concentrated solution obtained in the membrane separation step, ensuring the efficiency of the drying process and the quality of the product. Overall, this preparation method, through the close connection and parameter optimization of five steps—pretreatment, ultrasonic-assisted extraction, compound enzymatic hydrolysis, membrane separation, and spray drying—forms a complete technology chain from raw material crushing, component release, selective purification to finished product drying. Each step works synergistically at the physical, chemical, and biological levels to achieve efficient, high-purity extraction and stable preparation of the active ingredients of *Adenophora stricta*.

[0043] This invention provides an application of an effective component extract of Adenophora stricta, which is applied to the field of improving lung health, specifically in the areas of treating lung inflammation, improving lung function, preventing lung diseases, and adjuvant treatment of lung diseases.

[0044] Specifically, in this embodiment of the invention, lung inflammation is treated with North American Ginseng Capsules.

[0045] Specifically, each capsule of North American ginseng contains 0.4g of active ingredient powder, including 100g of North American ginseng extract powder, 15g of dextrin, 25g of sorbitol, and 2g of magnesium stearate.

[0046] The ingredients and dosages of the North American Ginseng Capsules provided by this invention do not limit the technology. Other excipients can be added and the dosages of each ingredient can be adjusted according to actual needs.

[0047] The relevant detection methods involved in the embodiments and comparative examples of this invention are as follows: 1. Polysaccharide Detection of Active Components Extract from *Adenophora stricta*: Considering the high content of polysaccharides in *Adenophora stricta* and their key component focus in this invention, the determination of polysaccharides was used as the core indicator for exploring enzymatic hydrolysis process parameters to make the optimization of enzymatic hydrolysis conditions more targeted and practical. The polysaccharide detection method was the phenol-sulfuric acid method, the instrument was a UV-Vis spectrophotometer, and the detection standard was the Chinese Pharmacopoeia (2025 edition) 0401 UV-Vis spectrophotometry.

[0048] 2. Anti-inflammatory activity test of the active ingredient extract of North American ginseng: Given that inflammation permeates the pathological development of numerous diseases, the search for drugs with natural anti-inflammatory activities has long been a key objective in related research fields. In this experiment, to evaluate the anti-inflammatory activity of the extract of active ingredients from *Adenophora stricta* prepared in Example 1, researchers investigated the effect of this extract on RAW264.7 cells, using intracellular nitric oxide (NO) production as the core indicator. RAW264.7 cells were used, and the concentration was adjusted to 1×10⁻⁶. 5 Cells were seeded evenly in 96-well plates at a concentration of 100 cells / mL and cultured statically for 24 hours. Then, lipopolysaccharide (LPS) was added to treat the cells, thus constructing an inflammatory cell model. Two groups were set up: a blank control group received no inflammatory treatment, while the LPS positive control group received only LPS treatment throughout. Subsequently, the active ingredient extract of *Adenophora stricta* was diluted to five concentration gradients (2, 4, 6, 8, and 10 μg / mL) using DMEM (Durbeco modified Eagle medium) and added to the constructed inflammatory cell model. After 2 hours of culture, DMEM was added to the blank control group, while LPS was added to both the LPS positive control group and the experimental group at a concentration of 1 μg / mL. All samples were then cultured at 37°C and 5% CO2 for another 24 hours. After culture, the supernatant from each group was collected to determine the NO content. The detection results are shown in the figure below. Figure 6 As shown.

[0049] 3. Effects of Bei Sha Shen capsules on the PI3K / Akt signaling pathway and pulmonary inflammatory response in lung cancer rats. a. Fifteen Wistar rats were randomly selected from 90 Wistar rats as the control group, and the remaining 75 rats were used to establish a lung cancer model by intratracheal instillation of carcinogenic iodized oil solution. They were divided into the model group, chemotherapy drug group, low-dose North American ginseng extract group, medium-dose North American ginseng extract group, and high-dose North American ginseng extract group.

[0050] b. The control group and model group were given saline by gavage. The chemotherapy drug group was given temozolomide capsules (60 mg / kg) by gavage. The low-dose group, medium-dose group and high-dose group of North American ginseng extract were given North American ginseng capsules (100 mg / kg, 200 mg / kg and 400 mg / kg) prepared in Example 18 by gavage. The rats were sacrificed after two weeks of continuous administration.

[0051] (1) Detection of spleen and lung indices: 24 hours after the last drug administration, the following operations were performed on each group of rats to determine the spleen and lung indices: First, the total weight of each rat was accurately weighed and the data was recorded; then the rats were sacrificed and the complete spleen and lung tissues were quickly removed; the adipose tissue and mesentery attached to the surface of the organs were carefully removed and the blood stains on the surface of the organs were gently absorbed with clean filter paper to ensure accurate measurement of organ weight; the net weight of the processed spleen and lungs were weighed separately, and then the organ samples were transferred to a -80℃ low temperature environment for freezing and preservation for subsequent experiments; finally, the relevant organ indices were calculated according to the formula: the spleen index was obtained by "net weight of spleen ÷ weight of rat", and the lung index was obtained by "net weight of lung ÷ weight of rat".

[0052] (2) Immune function test: Blood samples were collected from each group of rats. 5 mL of blood was taken from each rat. After anticoagulation treatment of 2 mL of blood, the following immune cell-related indicators were analyzed by flow cytometry (flow cytometer detection): percentage of T lymphocyte surface markers CD3⁺ cells, percentage of CD4⁺ cells, and percentage of CD8⁺ cells. At the same time, the ratio of CD4⁺ cells to CD8⁺ cells (CD4⁺ / CD8⁺) was calculated and recorded.

[0053] (3) Detection of inflammatory factor levels: The blood sample collection method is the same as the above steps. After collecting 5 mL of blood from each rat, take 3 mL of blood sample for further processing: Centrifuge this part of blood at 3000 rpm for 5 minutes to separate serum components. Then, use enzyme-linked immunosorbent assay (ELISA) to detect the levels of three cytokines in serum: interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α). The entire detection process must be strictly followed according to the instructions of the kit used.

[0054] (4) PI3K / Akt expression detection: Pathological morphology observation of lung tissue. Sample processing: 50g of lung tissue from each group of rats was taken and fixed in 10% neutral formaldehyde solution. Paraffin embedding, sectioning, and HE (hematoxylin-eosin) staining were then performed. The pathological morphological changes of lung tissue from different groups of rats were observed under a microscope. At the same time, some lung tissue was frozen in liquid nitrogen for later use in subsequent experiments. PI3K / Akt mRNA expression level detection: Total RNA extraction: Total RNA was extracted from the frozen lung tissue samples using Trizol reagent. Reverse transcription: The quality of the extracted total RNA was assessed. After confirming that the sample was qualified, reverse transcription was performed to convert RNA into cDNA. Real-time quantitative PCR detection: The mRNA expression level of the PI3K / Akt gene was determined using real-time quantitative PCR technology. The relative gene expression level was determined by 2 -△△Ct The reaction conditions were set as follows: the PCR reaction was first pre-denatured at 95℃ for 30 seconds; then it entered the cycling stage and was carried out for a total of 40 cycles. Each cycle included three steps: denaturation at 95℃ for 10 seconds, annealing at 60℃ for 30 seconds, and extension at 72℃ for 30 seconds.

[0055] Example 1 The preparation of an extract of effective components from *Adenophora stricta* includes the following steps: S1. Pretreatment: Take fresh North American ginseng, peel it, dry it in a cool place, crush it, and pass it through a 10-mesh sieve to obtain North American ginseng powder for later use.

[0056] S2. Ultrasonic-assisted treatment: Take 400g of North American ginseng powder that has been pretreated in step S1, add purified water, and the ratio of purified water to North American ginseng powder is 30mL:1g. Extract for 1h under ultrasonic power of 300W and temperature of 60℃ to obtain ultrasonic-assisted treatment solution.

[0057] S3. Enzymatic hydrolysis: Cellulase, papain, and saccharifying enzyme are added to the ultrasonic-assisted treatment solution obtained in step S2. The total mass of cellulase, papain, and saccharifying enzyme is 3.0% of the mass of the North American ginseng powder taken in step S2. The mass ratio of cellulase, papain, and saccharifying enzyme is 3:1:1. The pH of the system is adjusted to 5, the enzymatic hydrolysis temperature is 60℃, and the enzymatic hydrolysis is carried out for 3 hours to obtain the enzymatic extract.

[0058] S4. Membrane separation: The enzymatic extract obtained in step S3 is ultrafiltered through an ultrafiltration membrane to obtain a concentrated solution containing the effective components of North American ginseng extract.

[0059] S5. Drying: The concentrate obtained in step S4 is spray-dried with an inlet air temperature of 180℃, a fan frequency of 45Hz, a feed flow rate of 17 mL / min, and an outlet air temperature of 80℃ to obtain the extract of active ingredients of Glehnia littoralis.

[0060] Example 2 The preparation of an extract of effective components from *Adenophora stricta* includes the following steps: S1. Pretreatment: Take fresh North American ginseng, peel it, dry it in a cool place, crush it, and pass it through an 8-mesh sieve to obtain North American ginseng powder for later use.

[0061] S2. Ultrasonic-assisted treatment: Take 400g of North American ginseng powder that has been pretreated in step S1, add purified water, and the ratio of purified water to North American ginseng powder is 35mL:1g. Extract for 2h under ultrasonic power of 350W and temperature of 70℃ to obtain ultrasonic-assisted treatment solution.

[0062] S3. Enzymatic hydrolysis: Cellulase, papain, and saccharifying enzyme are added to the ultrasonic-assisted treatment solution obtained in step S2. The total mass of cellulase, papain, and saccharifying enzyme is 2.0% of the mass of the North American ginseng powder taken in step S2. The mass ratio of cellulase, papain, and saccharifying enzyme is 5:1:1. The pH of the system is adjusted to 5, the enzymatic hydrolysis temperature is 65℃, and the enzymatic hydrolysis is carried out for 3 hours to obtain the enzymatic hydrolysis extract.

[0063] S4. Membrane separation: The enzymatic extract obtained in step S3 is ultrafiltered through an ultrafiltration membrane to obtain a concentrated solution containing the effective components of North American ginseng extract.

[0064] S5. Drying: The concentrate obtained in step S4 is spray-dried with an inlet air temperature of 160℃, a fan frequency of 50Hz, a feed flow rate of 15mL / min, and an outlet air temperature of 80℃ to obtain the extract of active ingredients of Glehnia littoralis.

[0065] Example 3 The preparation of an extract of effective components from *Adenophora stricta* includes the following steps: S1. Pretreatment: Take fresh North American ginseng, peel it, dry it in a cool place, crush it, and pass it through a 12-mesh sieve to obtain North American ginseng powder for later use.

[0066] S2. Ultrasonic-assisted treatment: Take 400g of North American ginseng powder that has been pretreated in step S1, add purified water, and the ratio of purified water to North American ginseng powder is 25mL:1g. Extract for 1h under ultrasonic power of 250W and temperature of 50℃ to obtain ultrasonic-assisted treatment solution.

[0067] S3. Enzymatic hydrolysis: Cellulase, papain, and saccharifying enzyme were added to the ultrasonic-assisted treatment solution obtained in step S2. The total mass of cellulase, papain, and saccharifying enzyme was 4.0% of the mass of the North American ginseng powder taken in step S2. The mass ratio of cellulase, papain, and saccharifying enzyme was 2.5:1:1. The pH of the system was adjusted to 6, the enzymatic hydrolysis temperature was 55℃, and the enzymatic hydrolysis was carried out for 3.5 hours to obtain the enzymatic extract.

[0068] S4. Membrane separation: The enzymatic extract obtained in step S3 is ultrafiltered through an ultrafiltration membrane to obtain a concentrated solution containing the effective components of North American ginseng extract.

[0069] S5. Drying: The concentrate obtained in step S4 is spray-dried with an inlet air temperature of 200℃, a fan frequency of 35Hz, a feed flow rate of 20mL / min, and an outlet air temperature of 80℃ to obtain the extract of active ingredients of Glehnia littoralis.

[0070] Example 4 The preparation of an extract of effective components of North American ginseng, in this Example 4, adopts the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis time is 1 hour.

[0071] Example 5 The preparation of an extract of effective components of Glehnia littoralis, in this Example 5, adopts the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis time is 2 hours.

[0072] Example 6 The preparation of an extract of effective components of North American ginseng, in this Example 6, adopts the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis time is 4 hours.

[0073] The changes in polysaccharide extraction rate between Example 1 and Examples 4-6 are shown in the graph. Figure 1 As shown.

[0074] according to Figure 1 As shown in Table 1, the extraction rate of *Adenophora stricta* polysaccharides is affected by the enzymatic hydrolysis time. During the enzymatic hydrolysis process, the extraction rate first increases and then decreases with the extension of the hydrolysis time. The extraction rate reaches its highest value when the hydrolysis time is 3 hours. This is because, in Examples 4 and 5, when the hydrolysis time is short, the enzymatic hydrolysis has not been fully developed. As the hydrolysis time gradually increases, the enzymatic hydrolysis effect is continuously enhanced, thereby increasing the extraction rate of *Adenophora stricta* polysaccharides. However, when the hydrolysis time continues to be extended beyond 3 hours, in Example 6, some polysaccharides may be over-hydrolyzed, leading to a decrease in the extraction rate.

[0075] Example 7 The preparation of an extract of effective components of Glehnia littoralis, in this Example 7, adopts the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis temperature is 40℃.

[0076] Example 8 The preparation of an extract of effective components of Glehnia littoralis, in this Example 8, uses the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis temperature is 50°C.

[0077] Example 9 The preparation of an extract of effective components of Glehnia littoralis, in this Example 9, adopts the same method as in Example 1, except that: in step S3, the enzymatic hydrolysis temperature is 70℃.

[0078] The graph showing the change in polysaccharide extraction rate between Example 1 and Examples 7-9 is shown below. Figure 2 As shown.

[0079] according to Figure 2 As shown in Table 1, the relationship between enzymatic hydrolysis temperature and the extraction rate of polysaccharides from *Adenophora stricta* exhibits a clear phased characteristic: within the temperature range of 40–60℃, the polysaccharide extraction rate gradually increases with rising temperature; however, when the temperature exceeds 60℃, the extraction rate decreases significantly with further increases in temperature, reaching its experimental peak at 60℃. This is because, within the 40–60℃ range, the increase in temperature has two positive effects: firstly, it enhances the solubility of polysaccharides in the extraction solvent, allowing more polysaccharides to dissolve from the raw material into the solvent; secondly, it increases the diffusion coefficient of polysaccharide molecules, accelerating their diffusion rate from the raw material into the solvent. Furthermore, this temperature range also matches the optimal operating temperature of saccharifying enzymes, resulting in stable enzyme activity and efficient conversion of insoluble carbohydrates. These multiple factors collectively drive the increase in extraction rate. However, when the temperature exceeds 60℃, excessively high temperatures can cause negative effects. First, they can directly damage the molecular structure of polysaccharides (such as breaking glycosidic bonds and changing spatial conformation), leading to the inactivation or degradation of some of the dissolved polysaccharides. Second, they can exceed the tolerance temperature range of saccharifying enzymes, causing the enzyme protein to denature and become inactive, thus preventing it from continuing to perform enzymatic hydrolysis. In some cases, the destruction of the enzyme structure may even produce impurities, further affecting the polysaccharide extraction effect and ultimately leading to a decrease in the extraction rate.

[0080] Example 10 The preparation of an extract of effective components of Glehnia littoralis, in this Example 10, adopts the same method as in Example 1, except that in step S2, the ratio of purified water to Glehnia littoralis powder is 20 mL: 1 g.

[0081] Example 11 The preparation of an extract of effective components of Glehnia littoralis, in this Example 10, uses the same method as in Example 1, except that in step S2, the ratio of purified water to Glehnia littoralis powder is 40 mL: 1 g.

[0082] Example 12 The preparation of an extract of effective components of Glehnia littoralis, in this Example 10, uses the same method as in Example 1, except that in step S2, the ratio of purified water to Glehnia littoralis powder is 50 mL: 1 g.

[0083] The graph showing the change in polysaccharide extraction rate between Example 1 and Examples 10-12 is shown below. Figure 3 As shown.

[0084] according to Figure 3 As shown in Table 1, the liquid-to-solid ratio has a significant impact on the extraction rate of *Adenophora stricta* polysaccharides. During the experiment, the extraction rate of *Adenophora stricta* polysaccharides reached its peak when the liquid-to-solid ratio reached 30:1 (mL / g). However, as the liquid-to-solid ratio continued to increase, the extraction rate showed a decreasing trend. This phenomenon is because when the liquid-to-solid ratio is too high, the amount of solvent is relatively excessive, which is not conducive to the full dissolution of *Adenophora stricta* polysaccharides from the raw material, leading to a decrease in the extraction rate.

[0085] Example 13 The preparation of an effective component extract of Glehnia littoralis, in this Example 13, adopts the same method as in Example 1, except that: in step S3, the total mass of cellulase, papain and saccharifying enzyme is 1.0% of the mass of Glehnia littoralis powder taken in step S2.

[0086] Example 14 The preparation of an effective component extract of Glehnia littoralis, in this Example 14, adopts the same method as in Example 1, except that: in step S3, the total mass of cellulase, papain and saccharifying enzyme is 2.0% of the mass of Glehnia littoralis powder taken in step S2.

[0087] Example 15 The preparation of an effective component extract of Glehnia littoralis, in this Example 15, adopts the same method as in Example 1, except that: in step S3, the total mass of cellulase, papain and saccharifying enzyme is 4.0% of the mass of Glehnia littoralis powder taken in step S2.

[0088] The graph showing the change in polysaccharide extraction rate between Example 1 and Examples 13-15 is shown below. Figure 4 As shown.

[0089] according to Figure 4As shown in Table 1, the total amount of enzymes added and the extraction rate of North American ginseng polysaccharides exhibit a trend of "first increasing and then decreasing." The extraction rate reached its highest level in the experiment when the total amount of cellulase, papain, and saccharifying enzymes added was 3%. This trend can be analyzed from two aspects: the "dose-effect of enzyme-catalyzed reactions" and the "risk of over-reaction." Within the range where the enzyme addition amount is less than 3%, as the enzyme dosage increases, the number of enzyme molecules that can participate in the reaction increases, allowing for more complete contact with the substrates in the North American ginseng raw material. On the one hand, this efficiently hydrolyzes the plant cell wall structure and intracellular insoluble carbohydrates (such as starch and cellulose), breaking down the physical barrier to polysaccharide dissolution; on the other hand, it accelerates the conversion of poorly soluble carbohydrates into soluble polysaccharides. This dual effect drives a gradual increase in the polysaccharide extraction rate. However, when the enzyme addition exceeds 3%, excessive cellulase, papain, and saccharifying enzyme can cause "over-enzymatic hydrolysis": in addition to acting on non-target substrates, the excess enzymes will further degrade the dissolved North American ginseng polysaccharides, destroy the molecular structure of the polysaccharides (such as breaking glycosidic bonds), and decompose them into oligosaccharides or monosaccharides whose degree of polymerization does not meet the detection standards, resulting in a reduction in the total amount of effective polysaccharides in the extract, which ultimately leads to a decrease in the extraction rate.

[0090] The polysaccharide extraction rates of the effective components of Glehnia littoralis prepared in Examples 1-15 are shown in Table 1.

[0091] project Polysaccharide extraction rate (%) project Polysaccharide extraction rate (%) project Polysaccharide extraction rate (%) Example 1 38.0 Example 6 28.1 Example 11 28.2 Example 2 28.4 Example 7 20.2 Example 12 29.1 Example 3 29.5 Example 8 28.1 Example 13 27.1 Example 4 28.1 Example 9 30.1 Example 14 28.2 Example 5 26.0 Example 10 26.0 Example 15 29.0

[0092] Example 16 This embodiment uses the same method as Example 1, except that in step S3, different experiments were conducted with different mass ratios of cellulase, papain, and saccharifying enzyme at (5:1:1), (4:1:1), (2:1:1), and (1:1:1), respectively, and these were recorded as serial numbers 1, 2, 3, and 4. The experimental results were compared with those of Example 1. The experimental design and results are shown in Table 2. The polysaccharide extraction rate variation graph in this embodiment is shown in Figure 1. Figure 5 As shown.

[0093] Table 2. Effects of different mass ratios of cellulase, papain, and saccharifying enzyme on polysaccharide extraction rate. Serial Number Mass ratio of cellulase, papain and saccharifying enzyme Polysaccharide extraction rate (%) 1 5：1：1 30.4 2 4：1：1 34.8 3 2：1：1 26.2 4 1：1：1 22.2 Example 1 3：1：1 38.0 according to Figure 5 As shown in Table 1, the functions of cellulase, papain, and saccharifying enzyme are complementary. Their synergistic effect can only be fully realized when they are combined in an appropriate mass ratio. If the ratio is unbalanced, one of the enzymes may not function effectively (e.g., incomplete cell wall disruption or insufficient protein removal), making it difficult to achieve a significant increase in extraction rate. Based on experimental results, the optimal mass ratio of cellulase, papain, and saccharifying enzyme was determined to be 3:1:1.

[0094] Example 17 This embodiment uses the same method as Example 1. In step S3, the pH of the system is adjusted to 5, the enzymatic hydrolysis temperature is 60℃, and the enzymatic hydrolysis is carried out for 3 hours. The difference is that in step S3, different enzymes and different mass ratios are selected to conduct different experiments, which are recorded as serial numbers 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, and 13 respectively. The experimental results are compared with those of Example 1. The experimental design and results are shown in Table 3.

[0095] Table 3. Effects of enzyme type and dosage on polysaccharide extraction rate Serial Number Enzyme types (mass ratio) Polysaccharide extraction rate (%) 1 No enzyme added 17.2 2 Cellulase 28.8 3 Papain 26.8 4 Glucoamylase 25.1 5 Cellulase and papain (3:1) 29.4 6 Cellulase and papain (2:1) 28.1 7 Cellulase and papain (1:1) 27.6 8 Cellulase and glucoamylase (3:1) 32.4 9 Cellulase and glucoamylase (2:1) 31.6 10 Cellulase and glucoamylase (1:1) 30.5 11 Papain and glucoamylase (3:1) 29.4 12 Papain and glucoamylase (2:1) 28.2 13 Papain and glucoamylase (1:1) 27.5 Example 1 Cellulase, papain, and glucoamylase (3:1:1) 38.0 According to the data in Tables 2 and 3, the preparation method of the effective component extract of Glehnia littoralis provided by the present invention has a high extraction rate of polysaccharide extract. The extraction rates of polysaccharides extracted by a single enzyme or by a mixture of two enzymes are lower than the experimental results of Example 1. When cellulase, papain and saccharifying enzyme work together and the amount of enzymes is precisely controlled, the extraction rate of polysaccharide extract can be increased to 38% under the synergistic effect of the extraction process of the present invention.

[0096] according to Figure 6 The data shows that the anti-inflammatory activity of the active ingredient extract of *Adenophora stricta* in Example 1 was as follows: the NO content in the negative control group of RAW264.7 cells was 12.06 ± 0.48 μmol / L. After constructing an inflammatory cell model using LPS, the NO content in this model increased to 66.41 ± 3.28 μmol / L, indicating that the construction of the inflammatory cell model in Example 1 met expectations and the modeling was successful. Under the intervention of the active ingredient extract of *Adenophora stricta*, the NO content in the cells gradually decreased with changes in the extract concentration. This phenomenon indicates that the extract has a good anti-inflammatory effect, and the anti-inflammatory activity shows a corresponding trend with increasing concentration; specifically, when the extract concentration reached 10 μg / mL, the NO production in the cells was 13.04 ± 0.61 μmol / L.

[0097] Example 18 Preparation of a North American Ginseng Capsule: (1) Take 100g of the North American Ginseng effective component extract powder prepared in Example 1, 15g of dextrin, 25g of sorbitol and 2g of magnesium stearate, and mix them evenly.

[0098] (2) The above mixture is prepared into capsules containing 0.4g each using conventional capsule filling process.

[0099] The results of spleen index, lung index, immune function, inflammatory factor levels, and PI3K / Akt expression detection are as follows: (1) Detection of spleen index and lung index: Compared to the control group, the spleen index and lung index of the other groups of rats showed opposite trends: in terms of spleen index, the chemotherapy drug group, the high-dose group of North American ginseng extract, the medium-dose group of North American ginseng extract, the low-dose group of North American ginseng extract and the model group all showed significant decreases; in terms of lung index, these five groups showed sequential increases. For details, please refer to Table 4.

[0100] Table 4. Comparison of spleen index and lung index among the six groups of rats (x±s) Group Spleen index Lung Index control group 3.16±0.28 4.26±0.54 Model group 1.76±0.18 6.14±0.72 Chemotherapy drug group 2.78±0.34 4.46±0.54 Low-dose group of North American ginseng extract 2.06±0.29 5.24±0.26 Medium-dose group of North American ginseng extract 2.34±0.32 4.83±0.32 High-dose group of North American ginseng extract 2.47±0.35 4.64±0.34 (2) Results of rat immune function testing: Compared with the control group, the proportions of relevant immune cell subsets in the other five groups of rats (chemotherapy group, high-dose North American ginseng extract group, medium-dose North American ginseng extract group, low-dose North American ginseng extract group, and model group) showed a consistent trend: CD3 + T lymphocyte percentage, CD4 + T lymphocyte percentage and CD4 + / CD8 + The T lymphocyte ratios all showed a sequential decreasing trend, while the CD8 ratios... + The percentage of T lymphocytes shows an increasing trend, as detailed in Table 5.

[0101] Table 5. Comparison of immune function among the six groups of rats (x±s) Group <![CDATA[CD3 + (%)]]> <![CDATA[CD4 + (%)]]> <![CDATA[CD8 + (%)]]> <![CDATA[CD4 + / CD8 + (%)]]> control group 66.28±3.44 44.52±2.23 22.22±1.35 2.00±0.29 Model group 44.06±2.22 32.60±2.08 33.35±1.67 0.98±0.16 Chemotherapy drug group 56.88±2.42 40.22±2.08 25.28±1.60 1.59±0.21 Low-dose group of North American ginseng extract 50.43±1.49 34.55±2.04 27.33±1.29 1.26±0.18 Medium-dose group of North American ginseng extract 52.85±1.92 35.66±2.15 26.69±1.48 1.34±0.13 High-dose group of North American ginseng extract 54.62±1.87 38.91±2.17 25.32±1.24 1.54±0.15 (3) Detection of inflammatory factor levels in rats: Compared to the control group, the levels of three inflammation-related cytokines in the other five groups (chemotherapy group, high-dose North American ginseng extract group, medium-dose North American ginseng extract group, low-dose North American ginseng extract group, and model group) showed a consistent trend: the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) increased sequentially with the group. See Table 6.

[0102] Table 6. Comparison of inflammatory factor levels in six groups of rats (x±s, pg / mL) Group IL-6 IL-1β TNF-α control group 1.62±0.28 33.21±1.68 14.06±1.32 Model group 5.55±0.82 128.66±2.96 47.44±1.85 Chemotherapy drug group 2.89±0.26 58.47±2.68 26.42±1.42 Low-dose group of North American ginseng extract 3.81±0.77 69.82±2.74 37.44±1.72 Medium-dose group of North American ginseng extract 3.64±0.68 64.39±2.61 33.26±1.62 High-dose group of North American ginseng extract 3.56±0.76 62.35±2.31 31.33±1.53 (4) Detection of PI3K / Akt expression in 6 groups of rats Compared with the control group, the expression levels of PI3K and Akt in the model group, the high-dose group of North American ginseng extract, the medium-dose group of North American ginseng extract, the low-dose group of North American ginseng extract, and the chemotherapy drug group showed a gradual upward trend. The results are shown in Table 7.

[0103] Table 7. Comparison of PI3K / Akt expression levels in six groups of rats (x±s) Group PI3K Akt control group 0.09±0.02 0.15±0.03 Model group 0.91±0.21 1.22±0.36 Chemotherapy drug group 0.49±0.17 0.68±0.23 Low-dose group of North American ginseng extract 0.66±0.18 0.74±0.25 Medium-dose group of North American ginseng extract 0.60±0.16 0.71±0.23 High-dose group of North American ginseng extract 0.58±0.15 0.70±0.21 In summary, significant differences were observed in multiple indicators among different groups: compared with the control group, the spleen index and CD3 levels were significantly higher in the chemotherapy drug group, the high-dose North American ginseng group, the medium-dose North American ginseng group, the low-dose North American ginseng group, and the model group. + CD4 + Cell percentage and CD4 + / CD8 + The ratio showed a gradual decreasing trend, while the lung index and CD8... + The percentage of cells and the expression levels of IL-6, IL-1β, and TNF-α in serum showed a gradual increasing trend. This phenomenon indicates that Glehnia littoralis can effectively regulate the organ indices of lung cancer rats, promote the improvement of their immune function, and inhibit the occurrence and development of inflammatory responses. The regulation of the PI3K / Akt signaling pathway is an important pathway for Glehnia littoralis to achieve these effects.

[0104] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are exhaustively listed. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0105] For those skilled in the art, various modifications and improvements can be made without departing from the concept of the present invention, and these modifications and improvements are all within the scope of protection of the present invention. The scope of protection of the present invention is defined by the appended claims.

Claims

1. A preparation method of an effective component extract of Radix Glycymhizae, characterized in that, The preparation method comprises the following steps: S1, pretreatment: taking radix glabrae, drying, crushing, and sieving to obtain radix glabrae powder for standby; S2, ultrasonic-assisted treatment: taking the radix glabrae powder after the pretreatment in step S1, adding water, and performing extraction under the action of ultrasonic waves to obtain an ultrasonic-assisted treatment solution; S3, enzymatic hydrolysis treatment: adding cellulase, papain, and glucoamylase to the ultrasonic-assisted treatment solution obtained in step S2, adjusting the pH of the system to 5-6 for enzymatic hydrolysis to obtain an enzymatic hydrolysis extract; S4, membrane separation: filtering the enzymatic hydrolysis extract obtained in step S3 to obtain a concentrated solution containing radix glabrae active ingredient extract; S5, drying: performing spray drying on the concentrated solution obtained in step S4 to obtain a radix glabrae active ingredient extract.

2. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S1, the powder is sieved through an 8-12 mesh sieve. In step S2, the ratio of water to radix glabrae powder is (20-50) mL:1 g.

3. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S4, the filtration is ultrafiltration, and an ultrafiltration membrane with a molecular weight cut-off of 3.0 kDa is used for ultrafiltration.

4. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S2, the ultrasonic wave power is 250-350 W. ​ 5. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S2, the temperature is 50-70 DEG C, and the extraction time is 0.5-2 h. ​ 6. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S3, the enzymatic hydrolysis temperature is 40-70 DEG C, and the enzymatic hydrolysis time is 1-4 h. ​ 7. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S3, the total mass of cellulase, papain, and glucoamylase is 1.0-4.0% of the mass of the radix glabrae powder taken in step S2, and the mass ratio of cellulase, papain, and glucoamylase is (2.5-5):1:

1.

8. The preparation method of the effective component extract of Radix Glycyrhizae according to claim 1, characterized in that, In step S5, the inlet air temperature of the spray drying is 160-200 DEG C, the fan frequency is 35-50 Hz, and the feed flow rate is 15-20 mL / min; the outlet air temperature is 60-90 DEG C.

9. An extractive of effective components of Radix Glycyrrhizae, characterized in that, The radix glabrae active ingredient extract is prepared according to the preparation method of any one of claims 1-8.

10. The use of the effective component extract of Radix Glycyrhizae according to claim 9, characterized in that, The radix glabrae active ingredient extract is applied to the field of improving lung health.

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