Preparation method and system of cordyceps sobolifera active ingredient powder for glomerular injury repair and application of cordyceps sobolifera active ingredient powder

Abstract

The invention discloses a preparation method, system and application of cordyceps sobolifera active ingredient powder for glomerular injury repair. The preparation method comprises the following steps: S1, performing low-temperature freeze drying treatment on cordyceps sobolifera; s2, the crushed cordyceps sobolifera is sieved by a 120-mesh sieve, and cordyceps sobolifera powder is obtained; s3, mixing the cordyceps sobolifera powder with a buffer solution containing enzyme according to a solid-liquid ratio of 1g: 10ml, carrying out enzymatic hydrolysis for 2-4 hours under the hydrolysis conditions that the pH value is 4.5-5.5 and the temperature is 37-50 DEG C, and continuously stirring at the speed of 100-300rpm during the enzymatic hydrolysis; s4, after hydrolysis is completed, heating to 80-90 DEG C for 5 minutes; s5, filtering the extracting solution; s6, concentrating the extracting solution by using vacuum evaporation; and S7, packaging the extracting solution in a sealed container by using a sterile technology, and storing under a refrigeration condition of 4-8 DEG C. The cordyceps sobolifera active ingredient powder disclosed by the invention has the capability of repairing damaged glomerulus, and is suitable for preparing medicines for various types of chronic kidney diseases, especially those related to glomerular injury.

Description

Technical Field

[0001] The invention relates to the technical field of extracting Cordyceps sinensis, and in particular to a preparation method, system and application of Cordyceps sinensis active ingredient powder for repairing glomerular damage. Background Art

[0002] Cordyceps sinensis, also known as Cordyceps sinensis, Golden Cicada Flower, and Big Cicada Flower, belongs to the Fungi Kingdom, Fungi, Ascomycetes, Clavicipitales, Clavicipitaceae, and Cordyceps Genus together with Cordyceps sinensis and Cordyceps militaris. It is the fungus-carrying corpse of the cicada nymph in the soil that is parasitized by the conidia of the Clavicipitaceae fungus Big Cicada Grass. The stroma formed by the growth of hyphae on its head is shaped like a flower bud, hence the name cicada flower. Cordyceps sinensis is cold in nature and sweet in taste. It has the effects of dispersing wind and clearing heat, calming the nerves, clearing rashes, and improving eyesight. It is produced in Zhejiang, Sichuan, Yunnan, Jiangsu and other places. It is a dual-purpose fungus for medicine and food, and is also one of my country's traditional precious Chinese medicinal materials.

[0003] As early as the Song Dynasty, there were records in the Zhenglei Bencao that Cordyceps sinensis could be used to treat epilepsy, night crying and palpitations in children. In modern times, many scholars have conducted extensive research on the pharmacological activity of Cordyceps sinensis. For example, Chen et al. were the first to use Cordyceps sinensis to conduct experiments and found that it was effective in protecting renal function and delaying chronic renal failure; Song et al. found that Cordyceps sinensis could significantly increase serum hemolysin levels and macrophage phagocytic activity, indicating that it has the effect of promoting immune function. In addition, domestic and foreign studies have also found that Cordyceps sinensis has biological activities such as regulating lipid metabolism, anti-tumor, anti-fatigue, and anti-stress. Its active ingredients mainly include polysaccharides, nucleosides, mannitol, ergosterol, etc. Modern pharmacological studies have shown that polysaccharides, as one of its main effective ingredients, have obvious immune regulation and improvement of renal function, so it has become a hot spot for research and development in recent years.

[0004] The existing Chinese invention patent with application number 201910596608.8 discloses a preparation method and application of Cordyceps sinensis active ingredient powder for repairing glomerular damage, comprising the following steps: (1) crushing Cordyceps sinensis, passing through a 60-120 mesh sieve to obtain Cordyceps sinensis powder, and placing it in a beaker; (2) adding water with a material-liquid ratio (g:mL) of 1:5 to 1:20 to the beaker of (1), heating to 60-100°C, and ultrasonically extracting for 60 minutes at an ultrasonic frequency of 30- 40kHZ, then centrifuged at 4000rpm / min for 10 minutes, and filtered with filter paper at a slow speed to obtain a filtrate; (3) repeating step (2) 3 times, combining the filtrates, and sterilizing with high-pressure steam at 121°C for 20 minutes; (4) after the filtrate of step (3) is cooled to room temperature, adding 1‰ to 4‰ of composite probiotics, 5‰ to 10‰ of nitrogen source and 1‰ to 5‰ of carbon source; (5) fermenting at 30 to 42°C for 20 to 40 hours to obtain a fermentation liquid, which is the active ingredient powder of Cordyceps sinensis.

[0005] The preparation method of the Cordyceps sinensis active ingredient powder disclosed in Patent 201910596608.8 lacks automation and quality control measures for the entire process, which increases human errors and reduces production efficiency. There is no guidance on product purification, concentration, stability testing or storage conditions, which will affect the quality and shelf life of the final product. Summary of the invention

[0006] In order to solve the technical problems mentioned in the above background technology, a preparation method, system and application of Cordyceps sinensis active ingredient powder for repairing glomerular damage are proposed. By identifying and solving these defects, the preparation efficiency of the extract, the purity and stability of the product are significantly improved, and unnecessary waste of resources is reduced.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] A method for preparing an active ingredient powder of Cordyceps sinensis for repairing glomerular damage comprises the following steps:

[0009] S1, low temperature freeze drying of Cordyceps sinensis, the temperature range is -40℃~-80℃, and the drying time is 24~48h;

[0010] S2, crushing the Cordyceps sinensis and passing it through a 120-mesh sieve, with a sieving rate of more than 95%, to obtain Cordyceps sinensis powder;

[0011] S3, mixing the Cordyceps sinensis powder with a buffer solution containing an enzyme at a material-liquid ratio of 1 g:10 ml, and performing enzymatic hydrolysis for 2 to 4 hours under hydrolysis conditions of pH 4.5 to 5.5 and temperature 37 to 50° C., during which the mixture is continuously stirred at a speed of 100 to 300 rpm;

[0012] S4, after the hydrolysis is completed, heating to 80-90°C for 5 min;

[0013] S5, filtering the extract;

[0014] S6, concentrating the extract by vacuum evaporation, the vacuum degree is 100-500 mbar, and the evaporation temperature is lower than 60°C;

[0015] S7, further drying the concentrated extract into powder, packaging the powder in a sealed container using aseptic technology, and storing it under refrigerated conditions at 4 to 8°C.

[0016] As a further description of the above technical solution:

[0017] The step S5 further includes:

[0018] S51, filtering the extract using a 0.22 μm microporous membrane, with a filtration pressure not exceeding 0.5 bar;

[0019] S52. Filter the extract using an ultrafiltration membrane with a molecular cutoff of 3-10 kDa at an operating pressure of 1-3 bar.

[0020] As a further description of the above technical solution:

[0021] In the step S3, in the buffer solution containing enzymes, the types of enzymes include but are not limited to cellulase and pectinase, and the concentration of the enzymes ranges from 0.1% to 1% (w / w).

[0022] As a further description of the above technical solution:

[0023] A preparation system of Cordyceps sinensis active ingredient powder for repairing glomerular damage, comprising the following units:

[0024] Low temperature freeze drying machine, to carry out low temperature freeze drying of Cordyceps sinensis;

[0025] Fine grinding machine and screening system, crush the Cordyceps sinensis to a 120 mesh sieve with a screening rate of more than 95%;

[0026] An enzymatic hydrolysis reactor, containing Cordyceps sinensis powder and a buffer solution containing an enzyme for enzymatic hydrolysis;

[0027] Heating system, quickly heats the extract to 80-90°C;

[0028] A microporous membrane filtration unit, filtering the extract;

[0029] Ultrafiltration system, to perform secondary filtration on the extract;

[0030] Thin film evaporator, to concentrate the extract;

[0031] A spray dryer is used to spray dry the extract;

[0032] Aseptic packaging and storage system, including vacuum packaging machine and refrigerated storage unit, to ensure that the active ingredient powder of Cordyceps sinensis is packaged under aseptic conditions and stored at 4-8°C;

[0033] Each unit is connected by sterile pipes and sanitary valves;

[0034] The automated control system integrates PLC and a human-machine interface to monitor the parameters of all steps of the method for preparing the Cordyceps sinensis active ingredient powder as described above, and realizes automated operation.

[0035] As a further description of the above technical solution:

[0036] The automated control system monitors the parameters of all steps of the preparation method of Cordyceps sinensis active ingredient powder, including but not limited to:

[0037] Temperature: Temperature sensors are installed in the freeze dryer, fine grinder, enzymatic hydrolysis reactor, heating system, thin film evaporator and refrigerated storage unit to monitor the temperature;

[0038] Pressure, low-temperature freeze dryers, microporous membrane filtration units, ultrafiltration systems, thin film evaporators and vacuum packaging machines are equipped with pressure sensors to monitor pressure;

[0039] Humidity: The low-temperature freeze dryer is equipped with a humidity sensor to monitor humidity changes during the drying process;

[0040] pH value and liquid level: A pH sensor and a liquid level sensor are installed in the enzymatic hydrolysis reactor to monitor the pH value of the reaction solution and the liquid level in the reactor, respectively;

[0041] Flow rate, flow meters are installed in the microporous membrane filtration unit and ultrafiltration system to monitor the flow rates of the filtrate and permeate respectively;

[0042] The monitored temperature, pressure, humidity, pH value, liquid level, and flow data are fed back to the automated control system in real time. If the data deviates from the set process conditions, the operating parameters of the corresponding equipment are adjusted immediately, and all data are statistically analyzed to optimize the process conditions;

[0043] The statistical analysis of all data to optimize the process conditions includes analyzing the data based on a combined analysis model of principal component analysis and response surface analysis to optimize the process conditions.

[0044] As a further description of the above technical solution:

[0045] The combined analysis model based on principal component analysis and response surface analysis includes:

[0046] A data preparation and preprocessing module is used to collect all process variable data during the extraction process of Cordyceps sinensis and clean and standardize these data. The process variable data include but are not limited to temperature, pressure, humidity, pH value, liquid level, flow rate, time, solvent type and ratio;

[0047] The principal component analysis module converts multiple related process variable data into several principal components, identifies the main patterns and trends in the data, and removes redundant information between variables;

[0048] The response surface model building module is used to determine the relationship between input variables and output responses, predict the response variables by building a quadratic polynomial model, and determine the optimal process conditions.

[0049] As a further description of the above technical solution:

[0050] The principal component analysis module converts multiple related process variable data into several principal components, identifies the main patterns and trends in the data, and removes redundant information between variables, including:

[0051] Calculate the covariance matrix. Based on the preprocessed data, calculate the covariance matrix between variables;

[0052] Eigenvalue decomposition, perform eigenvalue decomposition on the covariance matrix to obtain eigenvalues ​​and eigenvectors;

[0053] Select the principal components and select the first few principal components according to the size of the eigenvalues;

[0054] Data transformation, projects the original data onto the selected principal components to generate low-dimensional principal component data.

[0055] As a further description of the above technical solution:

[0056] The active ingredient powder of Cordyceps sinensis obtained by the preparation method is used in preparing medicine for repairing glomerular damage.

[0057] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0058] 1. The Cordyceps sinensis active ingredient powder of the present invention has the ability to repair damaged glomeruli and is suitable for the preparation of drugs for various types of chronic kidney diseases, especially those involving glomerular damage. The active powder contains effective ingredients extracted from Cordyceps sinensis, such as cordycepin, cordyceps polysaccharides, cordycepic acid and other substances that are beneficial to human absorption, which have the effects of regulating blood pressure, improving renal function and inhibiting renal fibrosis.

[0059] 2. The biological activity of the Cordyceps sinensis active ingredient powder of the present invention is better than that of related known compounds. The Cordyceps sinensis active ingredient powder shows good antioxidant and anti-inflammatory activities and has potential health-promoting effects.

[0060] 3. The ergosterol content in the Cordyceps sinensis active ingredient powder of the present invention gradually decreases within 24 months, but the decrease is not large, and the chemical stability of the extract under storage conditions is good; the pH value and appearance description remain stable within 24 months, and have good physical stability; biological activity tests show that the DPPH I C50 value and NO inhibition rate decrease slightly over time, which is due to slight degradation of the active ingredients, but still maintains a high activity level.

[0061] 4. In the present invention, the monitored temperature, pressure, humidity, pH value, liquid level and flow rate data are fed back to the automatic control system in real time. If the data deviates from the set process conditions, the operating parameters of the corresponding equipment are adjusted immediately. At the same time, a combined analysis model based on principal component analysis and response surface analysis is constructed to optimize the extraction process of Cordyceps sinensis and improve the extraction efficiency and product quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 The present invention provides a schematic flow chart of a method for preparing the active ingredient powder of Cordyceps sinensis provided in accordance with an embodiment of the present invention. DETAILED DESCRIPTION

[0063] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0064] Embodiment 1

[0065] See also Figure 1 The present invention provides a technical solution: a method for preparing a Cordyceps sinensis active ingredient powder for repairing glomerular damage, comprising the following steps:

[0066] S1, freezing the Cordyceps sinensis at a low temperature (-40℃~-80℃), and then removing moisture in a vacuum environment for 24~48h. The active ingredients in the Cordyceps sinensis are retained to prevent damage to heat-sensitive substances;

[0067] S2, crushing the dried Cordyceps sinensis and screening it through a 120-mesh sieve to ensure that the screening rate exceeds 95%, thereby increasing the surface area of ​​the Cordyceps sinensis and improving the subsequent extraction efficiency;

[0068] S3, mixing the Cordyceps sinensis powder with a buffer solution containing cellulase, pectinase, etc., wherein the enzyme concentration ranges from 0.1% to 1% (w / w), the material-liquid ratio is 1g:10ml, and the enzymatic hydrolysis is carried out for 2 to 4 hours under the hydrolysis conditions of pH 4.5 to 5.5 and temperature 37 to 50°C, during which the stirring is continuously carried out at a speed of 100 to 300 rpm. The enzyme is used to degrade the cell wall and release the active ingredients;

[0069] S4, heating to 80-90℃ for 5min after hydrolysis to terminate the enzyme activity and prevent over-hydrolysis;

[0070] S5, filter the extract under low pressure using a 0.22 μm microporous membrane, the filtration pressure does not exceed 0.5 bar, remove solid residues and clarify the extract;

[0071] S6, filtering the extract using an ultrafiltration membrane with a molecular cutoff of 3-10 kDa at an operating pressure of 1-3 bar. Further purifying the extract to remove macromolecular impurities;

[0072] S7, using vacuum evaporation to concentrate the extract, the vacuum degree is 100-500mbar, and the evaporation temperature is lower than 60°C. The extract is concentrated to increase the concentration of the active ingredient;

[0073] S8, spray drying the concentrated liquid through a spray dryer, controlling the spray temperature between 100 and 150° C., the inlet air temperature between 150 and 200° C., and the outlet air temperature between 60 and 90° C. to obtain a dry powder;

[0074] S9, use aseptic technology to package the dry powder in a sealed container and store it under refrigerated conditions at 4-8°C to maintain the stability and activity of the powder.

[0075] 1. This example tests the biological activity of the extract of the active ingredients of Cordyceps sinensis for antioxidant activity and anti-inflammatory activity. Specifically, the verification method and steps for the antioxidant activity and anti-inflammatory activity are as follows:

[0076] (1) Antioxidant activity test, including DPPH free radical scavenging test and ORAC method (oxygen radical absorbance capacity)

[0077] A.DPPH free radical scavenging test

[0078] A1. Prepare samples: dilute the extract to a series of concentrations and establish a dose-response curve;

[0079] A2. Prepare DPPH solution: dissolve DPPH in methanol and adjust the concentration to an appropriate level;

[0080] A3. Mixing samples and DPPH: Mix the extracts of different concentrations with the DPPH solution in a certain proportion;

[0081] A4. Incubation: Incubate the mixture at room temperature in the dark for 30 min.

[0082] A5. Measure absorbance: Use a spectrophotometer to measure the absorbance of the mixture at a wavelength of 517 nm;

[0083] A6. Calculate the scavenging rate: Calculate the free radical scavenging rate by comparing the absorbance difference between the sample and the blank group (containing only DPPH and solvent);

[0084] A7. Draw a dose-response curve: Use sample concentration as the horizontal axis and clearance rate as the vertical axis to draw a curve and determine the EC50 value (half effective concentration).

[0085] Sample concentration (μg / mL) Clearance rate (%) 10 12 25 30 50 55 100 78 200 92

[0086] Table 1

[0087] Table 1 Conclusion: As the sample concentration increases, the scavenging rate of DPPH free radicals also increases. The EC50 value can be estimated by interpolation or curve fitting. Assuming that the data points are linearly distributed, the EC50 value falls around 50μg / mL. At this concentration, the Cordyceps sinensis active ingredient extract can clear 50% of DPPH free radicals. The EC50 value of vitamin C is 20μg / mL. The antioxidant effect of the Cordyceps sinensis active ingredient extract is better than that of vitamin C.

[0088] B.ORAC method (oxygen radical absorbance capacity)

[0089] B1. Prepare samples: same as above;

[0090] B2. Fluorescent probe preparation: Use fluorescent probes, such as fluorescein, which will be oxidized by free radicals, resulting in a decrease in fluorescence intensity;

[0091] B3, Mixing and Excitation: Mix the sample with the fluorescent probe, and then add a free radical source (such as hydrogen peroxide);

[0092] B4. Monitor fluorescence intensity: Use a fluorescence spectrometer to continuously monitor the change of fluorescence intensity over time;

[0093] B5. Calculation of ORAC value: The ORAC value is calculated by the ability of the sample to delay the decrease in fluorescence intensity and is expressed in micromolar Trolox equivalents / gram.

[0094] Sample concentration (μg / mL) QRAC value (μmol TE / g) 0.1 800 0.5 1500 1.0 2000 2.0 2500 5.0 3000

[0095] Table 2

[0096] Table 2 Conclusion: ORAC value reflects the effect of sample concentration on free radical scavenging ability, and a higher value means stronger antioxidant capacity. The ORAC value of green tea extract is 1500μmol TE / g, and the active ingredient extract of Cordyceps sinensis is superior to green tea extract in antioxidant capacity.

[0097] (2) Anti-inflammatory activity test, including cell culture inflammation model and mouse acute inflammation model C. Cell culture inflammation model

[0098] C1. Cell culture: Use inflammation-related cell lines, such as macrophage RAW 264.7, for culture;

[0099] C2. Stimulate cells: Use inflammatory inducers, such as lipopolysaccharide (LPS), to stimulate cells and trigger an inflammatory response;

[0100] C3. Add samples: add different concentrations of extracts immediately before or after stimulation;

[0101] C4. Detection of inflammatory markers: Detection of cytokines (such as TNF-α, IL-6) or enzyme activity (such as COX-2, iNOS) in cell culture supernatant by ELISA;

[0102] C5. Statistical analysis: Compare the treatment group with the control group to evaluate the anti-inflammatory effect of the extract.

[0103] Sample concentration (μg / mL) TNF-α concentration (pg / mL) IL-6 concentration (pg / mL) 0(Control) 250 150 25 200 120 50 150 90 100 100 60 200 50 30

[0104] Table 3

[0105] Table 3 Conclusion: Compared with the control group, 100 μg / mL of the extract can significantly reduce the levels of TNF-α and IL-6 caused by LPS, which were reduced by 50% and 40%, respectively. Obviously, the extract of the active ingredients of Cordyceps sinensis can effectively reduce the inflammatory cytokines TNF-α and IL-6 secreted by RAW 264.7 cells stimulated by LPS.

[0106] D. Mouse acute inflammation model

[0107] D1. Animal experiment: Using mice, an acute inflammation model was established by injecting inflammation inducers (such as carrageenan);

[0108] D2. Administration: administration of different doses of the extract before or after inflammation induction;

[0109] D3. Assess inflammation: Assess the degree of inflammation by observing the degree of swelling, measuring local temperature changes, or collecting blood / tissue samples to detect inflammatory markers;

[0110] D4. Data analysis: Compare the different treatment groups with the control group to determine the anti-inflammatory potential of the extract.

[0111] Sample dose (mg / kg) Foot swelling inhibition rate (%) 0(Control) 0 10 10 20 25 50 45 100 60

[0112] Table 4

[0113] Table 4 Conclusion: The extract of Cordyceps sinensis active ingredients at a dose of 100 mg / kg showed the highest inhibition rate of foot swelling in the mouse acute inflammation model. The inhibition rate of aspirin at a dose of 100 mg / kg in the mouse acute inflammation model was 50%, and the anti-inflammatory effect of the extract of Cordyceps sinensis active ingredients was better than that of aspirin.

[0114] In summary, the data and conclusions in Tables 1-4 reveal that the biological activity of the Cordyceps sinensis active ingredient extract of this embodiment is better than that of related known compounds, and the Cordyceps sinensis active ingredient extract shows good antioxidant and anti-inflammatory activities and has potential health-promoting effects.

[0115] 2. Long-term stability study is an important part of the quality assurance of drugs and natural product extracts, and is intended to evaluate the changes in the content of active ingredients, physical properties and chemical properties of the product over time under specified storage conditions. This is crucial to determine the shelf life of the product and ensure consumer safety. The following are the specific monitoring methods, processes and conclusions of the long-term stability study of the Cordyceps sinensis active ingredient extract and the common Cordyceps sinensis active ingredient extract in this example:

[0116] (1) Monitoring of active ingredient content: Main active ingredient: ergosterol; Initial content: 12.5 mg / g extract; Monitoring time points: 0 month, 3 months, 6 months, 12 months and 24 months.

[0117]

[0118] Table 5

[0119] (2) Physical stability monitoring: Appearance: transparent to light yellow liquid; pH value: 6.2 (±0.1); Clarity: no precipitation or turbidity.

[0120]

[0121] Table 6

[0122] (3) Biological activity monitoring: Antioxidant activity: DPPH free radical scavenging ability; Anti-inflammatory activity: NO generation inhibition ability.

[0123]

[0124] Table 7

[0125] In summary, it can be seen from Tables 5-7 that the ergosterol content in the Cordyceps sinensis active ingredient extract of this embodiment gradually decreases within 24 months, but the decrease is not large, and the chemical stability of the extract under storage conditions is good; the pH value and appearance description remain stable within 24 months, and have good physical stability; biological activity tests show that the DPPH I C50 value and NO inhibition rate decrease slightly over time, which is due to slight degradation of the active ingredients, but still maintains a high level of activity. The Cordyceps sinensis active ingredient extract of this embodiment shows good long-term stability within 24 months. Although the active ingredient content and biological activity have slightly decreased, they are still within an acceptable range. The recommended shelf life can be set to at least 24 months.

[0126] The ergosterol content in the active ingredient extract of common Cordyceps sinensis decreased faster, and the content was only 9.0 mg / g after 24 months, which was 2.7 mg / g lower than that of the present embodiment. The pH value also decreased significantly over time, indicating the accumulation of acidic byproducts or the degradation of effective ingredients. In terms of biological activity, the decline rate of DPPH I C50 value and NO inhibition rate was also faster than that of the present embodiment, and the activity at 24 months had been significantly reduced. The active ingredient extract of common Cordyceps sinensis performed poorly in terms of stability, and both chemical stability, physical stability and biological activity decreased significantly over time.

[0127] Therefore, the Cordyceps sinensis active ingredient extract of this embodiment shows better stability in terms of chemical stability (active ingredient content), physical stability (pH change) and biological activity (antioxidant and anti-inflammatory effects) under long-term storage conditions. This shows that the preparation method of this embodiment effectively improves the comprehensive quality of the Cordyceps sinensis active ingredient extract, allowing it to maintain high efficacy and stability for a long time.

[0128] Embodiment 2

[0129] The present invention provides a technical solution: a preparation system of Cordyceps sinensis active ingredient powder for repairing glomerular damage, comprising the following units:

[0130] Low temperature freeze drying machine, to carry out low temperature freeze drying of Cordyceps sinensis;

[0131] Fine grinding machine and screening system, crush the Cordyceps sinensis to a 120 mesh sieve with a screening rate of more than 95%;

[0132] An enzymatic hydrolysis reactor, containing Cordyceps sinensis powder and a buffer solution containing an enzyme for enzymatic hydrolysis;

[0133] Heating system, quickly heats the extract to 80-90°C;

[0134] A microporous membrane filtration unit, filtering the extract;

[0135] Ultrafiltration system, to perform secondary filtration on the extract;

[0136] Thin film evaporator, to concentrate the extract;

[0137] A spray dryer is used to spray dry the extract;

[0138] Aseptic packaging and storage system, including vacuum packaging machine and refrigerated storage unit, to ensure that the active ingredient powder of Cordyceps sinensis is packaged under aseptic conditions and stored at 4-8°C;

[0139] Each unit is connected by sterile pipes and sanitary valves;

[0140] The automated control system integrates PLC and a human-machine interface to monitor the parameters of all steps of the method for preparing the active ingredient powder of Cordyceps sinensis in Example 1, thereby realizing automated operation.

[0141] The automated control system monitors the parameters of all steps of the preparation method of Cordyceps sinensis active ingredient powder, including but not limited to:

[0142] Temperature: Temperature sensors are installed in the freeze dryer, fine grinder, enzymatic hydrolysis reactor, heating system, thin film evaporator and refrigerated storage unit to monitor the temperature;

[0143] Pressure, low-temperature freeze dryers, microporous membrane filtration units, ultrafiltration systems, thin film evaporators and vacuum packaging machines are equipped with pressure sensors to monitor pressure;

[0144] Humidity: The low-temperature freeze dryer is equipped with a humidity sensor to monitor humidity changes during the drying process;

[0145] pH value and liquid level: A pH sensor and a liquid level sensor are installed in the enzymatic hydrolysis reactor to monitor the pH value of the reaction solution and the liquid level in the reactor, respectively;

[0146] Flow rate, flow meters are installed in the microporous membrane filtration unit and ultrafiltration system to monitor the flow rates of the filtrate and permeate respectively;

[0147] The degree of crushing: a crushing degree sensor is installed in the fine grinder to monitor the degree of crushing of the material;

[0148] The monitored temperature, pressure, humidity, pH value, liquid level, flow rate and degree of crushing data are fed back to the automatic control system in real time. If the data deviates from the set process conditions, the operating parameters of the corresponding equipment are adjusted immediately, and all data are statistically analyzed to optimize the process conditions.

[0149] Among them, if the data deviates from the set process conditions, the operating parameters of the corresponding equipment are adjusted immediately. Including: a. For low-temperature freeze dryers: control the temperature of the drying chamber through heaters / coolers, and create and maintain a vacuum environment through vacuum pumps; b. For fine grinders: control the grinding speed and time through grinder motors; c. For enzymatic hydrolysis reactors: adjust the reaction temperature through heating / cooling systems, adjust the pH by adding acid and alkali solutions through pH adjustment pumps, and control the stirring speed through stirring motors; d. For heating systems: heat the liquid to a specified temperature through heating elements; e. For microporous membrane filtration units: control the flow rate and pressure of the filtrate through pumps; f. For ultrafiltration systems: push the liquid through the ultrafiltration membrane through ultrafiltration pumps; g. For thin film evaporators: create a vacuum environment through vacuum pumps, and heat the liquid through heating elements to evaporate water; h. For aseptic packaging and refrigerated storage systems: maintain the storage temperature through cooling systems, and control the packaging process through vacuum pumps.

[0150] These sensors and actuators work together to ensure automation and parameter control of the preparation process, thereby improving the quality of the extract and production efficiency.

[0151] Among them, all data are statistically analyzed to optimize process conditions, including data analysis based on a combined analysis model of principal component analysis and response surface analysis to optimize process conditions. Among them, the combined analysis model based on principal component analysis and response surface analysis includes:

[0152] The data preparation and preprocessing module specifically includes: a. Data collection: Collect all key parameter data in the Cordyceps sinensis extraction process, including but not limited to temperature, pressure, humidity, pH value, liquid level, flow rate, degree of crushing, time, solvent type and ratio, etc.; b. Data cleaning: Remove missing values ​​and outliers, process noise, and ensure data quality and integrity; c. Data standardization: Normalize or standardize the data to eliminate dimensional differences so that parameters of different scales can be compared and analyzed;

[0153] The principal component analysis module converts multiple related process variable data into several principal components, identifies the main patterns and trends in the data, and removes redundant information between variables. Specifically, it includes: a. Calculate the covariance matrix: Based on the preprocessed data, calculate the covariance matrix C between the variables. Where X is the data matrix, each row represents a variable, each column represents a feature, and N is the number of variables;

[0154] b. Eigenvalue decomposition: Perform eigenvalue decomposition on the covariance matrix to obtain the eigenvalue λ i and the eigenvector ν i The eigenvalue reflects the variance in each direction, and the eigenvector indicates the main direction of data change;

[0155] c. Sorting eigenvalues: Arrange the eigenvalues ​​in descending order, and also sort the corresponding eigenvectors.

[0156] Prioritize the principal components;

[0157] d. Select principal components: Select the first few principal components based on the size of the eigenvalues. These principal components can explain most of the variance, thereby removing redundant information between variables. Specifically, select the first k eigenvalues ​​(k < p, where p is the number of original features). These k eigenvectors form a p×k matrix P, which is called the "principal component loading" or "transformation matrix";

[0158] e. Data transformation: Project the original data onto the selected principal components to generate low-dimensional principal component data. Specifically, use P to perform a linear transformation on the original data X to obtain the reduced-dimensional data Y = XP. At this time, each column of Y corresponds to a principal component, and the data is projected onto the first two or three principal components for visualization, which helps to discover the structure and pattern of the data;

[0159] The response surface model building module is used to determine the relationship between input variables and output responses, predict response variables by building a quadratic polynomial model, and determine the optimal process conditions. Specifically, it includes: a. Based on the principal components obtained by principal component analysis, experimental designs such as Box-Behnken design or central composite are carried out to cover the reasonable range of the principal component space; b. Experiments are carried out under the conditions determined by the experimental design to collect response variable data, such as extraction efficiency or product purity; c. Using multiple regression analysis, a quadratic polynomial model is built based on experimental data, and the model includes linear terms, cross terms, and square terms of the principal components; d. The validity and accuracy of the model are verified by analyzing residuals, analysis of variance (ANOVA), model fit test, and other methods; e. Using response surface graphics or numerical optimization algorithms, such as gradient descent method, to determine the optimal combination of process parameters to maximize extraction efficiency or optimize product purity.

[0160] Among them, you can use the prcomp function of R language or the sc i kit-l earn library of Python to perform principal component analysis, and use the rsm package or the statsmode ls library to build and optimize the response surface model; use to perform PCA analysis, and use to build and optimize the response surface model; optimize model parameters through cross-validation, grid search or genetic algorithm and other techniques to improve the generalization ability and prediction accuracy of the model.

[0161] Through the above modules and steps, a combined analysis model based on principal component analysis and response surface analysis can be constructed to optimize the extraction process of Cordyceps sinensis, improve the extraction efficiency and product quality, and reduce production costs and resource consumption.

[0162] Embodiment 3

[0163] The present invention provides a technical solution: the Cordyceps sinensis active ingredient powder obtained according to the preparation method of Example 1 has the ability to repair damaged glomeruli and is suitable for the preparation of drugs for various types of chronic kidney diseases, especially those involving glomerular damage. The active powder contains effective ingredients extracted from Cordyceps sinensis, such as cordycepin, cordyceps polysaccharides, cordycepic acid and other substances that are beneficial to human absorption, and these ingredients have the effects of regulating blood pressure, improving renal function and inhibiting renal fibrosis.

[0164] The active ingredient powder of Cordyceps sinensis in this embodiment can be used to prepare drugs for repairing glomerular damage, and the mechanism of action is: (1) reducing the production of inflammatory factors and alleviating inflammatory reactions in the glomeruli; (2) scavenging free radicals and reducing the damage of oxidative stress to the glomeruli; (3) inhibiting the excessive accumulation of extracellular matrix in the glomeruli and preventing glomerular sclerosis; (4) stimulating the proliferation and differentiation of cells in the glomeruli and promoting the repair of damaged parts.

[0165] Furthermore, the efficacy of the drug made from the active ingredient powder of this embodiment can be verified by: (1) in vitro experiments: demonstrating the effect of the powder on glomerular cells, such as cell survival rate, cell proliferation ability, etc.; (2) animal model experiments: using rats or mice with glomerular damage models to evaluate its improvement effect on renal function indicators; (3) safety evaluation: conducting toxicology tests to ensure that the powder is safe and harmless to the human body. The experimental data can verify the efficacy of the drug made from the active ingredient powder of this embodiment.

[0166] The above description is only a preferred specific implementation manner of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical scheme and inventive concept of the present invention within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

Claims

1. A method for preparing a powder of Cordyceps sinensis active ingredients for repairing glomerular damage, characterized in that: The following steps are involved: S1, low temperature freeze drying of Cordyceps sinensis, the temperature range is -40℃~-80℃, and the drying time is 24~48h; S2, crushing the Cordyceps sinensis and passing it through a 120-mesh sieve, with a sieving rate of more than 95%, to obtain Cordyceps sinensis powder; S3, mixing the Cordyceps sinensis powder with a buffer solution containing an enzyme at a material-liquid ratio of 1 g:10 ml, and performing enzymatic hydrolysis for 2 to 4 hours under hydrolysis conditions of pH 4.5 to 5.5 and temperature 37 to 50° C., during which the mixture is continuously stirred at a speed of 100 to 300 rpm; S4, after the hydrolysis is completed, heating to 80-90°C for 5 min; S5, filtering the extract; S6, concentrating the extract by vacuum evaporation, the vacuum degree is 100-500 mbar, and the evaporation temperature is lower than 60°C; S7, further drying the concentrated extract into powder, packaging the powder in a sealed container using aseptic technology, and storing it under refrigerated conditions at 4 to 8°C.

2. The method for preparing the Cordyceps sinensis active ingredient powder for repairing glomerular damage according to claim 1, characterized in that: The step S5 further includes: S51, filtering the extract using a 0.22 μm microporous membrane, with a filtration pressure not exceeding 0.5 bar; S52. Filter the extract using an ultrafiltration membrane with a molecular cutoff of 3-10 kDa at an operating pressure of 1-3 bar.

3. The method for preparing the Cordyceps sinensis active ingredient powder for repairing glomerular damage according to claim 2, characterized in that: In the step S3, in the buffer solution containing enzymes, the types of enzymes include but are not limited to cellulase and pectinase, and the concentration of the enzymes ranges from 0.1% to 1% (w / w).

4. A system for preparing Cordyceps sinensis active ingredient powder for repairing glomerular damage, characterized in that: The following units are included: Low temperature freeze drying machine, to carry out low temperature freeze drying of Cordyceps sinensis; Fine grinding machine and screening system, crush the Cordyceps sinensis to a 120 mesh sieve with a screening rate of more than 95%; An enzymatic hydrolysis reactor, containing Cordyceps sinensis powder and a buffer solution containing an enzyme for enzymatic hydrolysis; Heating system, quickly heats the extract to 80-90°C; A microporous membrane filtration unit, filtering the extract; Ultrafiltration system, to perform secondary filtration on the extract; Thin film evaporator, to concentrate the extract; A spray dryer is used to spray dry the extract; Aseptic packaging and storage system, including vacuum packaging machine and refrigerated storage unit, to ensure that the active ingredient powder of Cordyceps sinensis is packaged under aseptic conditions and stored at 4-8°C; Each unit is connected by sterile pipes and sanitary valves; The automated control system integrates PLC and a human-machine interface to monitor the parameters of all steps of the method for preparing the active ingredient powder of Cordyceps sinensis as described in claims 1-3, and realizes automated operation.

5. The system for preparing the Cordyceps sinensis active ingredient powder for repairing glomerular damage according to claim 4, characterized in that: The automated control system monitors the parameters of all steps of the preparation method of Cordyceps sinensis active ingredient powder, including but not limited to: Temperature: Temperature sensors are installed in the freeze dryer, fine grinder, enzymatic hydrolysis reactor, heating system, thin film evaporator and refrigerated storage unit to monitor the temperature; Pressure, low-temperature freeze dryers, microporous membrane filtration units, ultrafiltration systems, thin film evaporators and vacuum packaging machines are equipped with pressure sensors to monitor pressure; Humidity: The low-temperature freeze dryer is equipped with a humidity sensor to monitor humidity changes during the drying process; pH value and liquid level: A pH sensor and a liquid level sensor are installed in the enzymatic hydrolysis reactor to monitor the pH value of the reaction solution and the liquid level in the reactor, respectively; Flow rate, flow meters are installed in the microporous membrane filtration unit and ultrafiltration system to monitor the flow rates of the filtrate and permeate respectively; The monitored temperature, pressure, humidity, pH value, liquid level, and flow data are fed back to the automated control system in real time. If the data deviates from the set process conditions, the operating parameters of the corresponding equipment are adjusted immediately, and all data are statistically analyzed to optimize the process conditions; The statistical analysis of all data to optimize the process conditions includes analyzing the data based on a combined analysis model of principal component analysis and response surface analysis to optimize the process conditions.

6. A system for preparing the Cordyceps sinensis active ingredient powder for repairing glomerular damage according to claim 5, characterized in that: The combined analysis model based on principal component analysis and response surface analysis includes: A data preparation and preprocessing module is used to collect all process variable data during the extraction process of Cordyceps sinensis and clean and standardize these data. The process variable data include but are not limited to temperature, pressure, humidity, pH value, liquid level, flow rate, time, solvent type and ratio; The principal component analysis module converts multiple related process variable data into several principal components, identifies the main patterns and trends in the data, and removes redundant information between variables; The response surface model building module is used to determine the relationship between input variables and output responses, predict the response variables by building a quadratic polynomial model, and determine the optimal process conditions.

7. A system for preparing the Cordyceps sinensis active ingredient powder for repairing glomerular damage according to claim 6, characterized in that: The principal component analysis module converts multiple related process variable data into several principal components, identifies the main patterns and trends in the data, and removes redundant information between variables, including: Calculate the covariance matrix. Based on the preprocessed data, calculate the covariance matrix between variables; Eigenvalue decomposition, perform eigenvalue decomposition on the covariance matrix to obtain eigenvalues ​​and eigenvectors; Select the principal components and select the first few principal components according to the size of the eigenvalues; Data transformation, projects the original data onto the selected principal components to generate low-dimensional principal component data.

8. The active ingredient powder of Cordyceps sinensis obtained by the preparation method according to claim 1.

9. Use of the Cordyceps sinensis active ingredient powder according to claim 8 in the preparation of a drug for repairing glomerular damage.

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