Plant embryo extract extraction method based on high-quality plant germs

Abstract

The invention relates to the technical field of separation and extraction, in particular to a plant embryo extract extraction method based on high-quality plant germs.The method comprises the steps that corn germs are added into a solvent according to the initial solid-to-liquid ratio, and graded smashing and water extraction are conducted; defining the germination degree to determine the germination state of the current batch of corn germs; determining an initial crushing parameter of graded crushing, predicting the viscosity change of the suspension after primary crushing, and determining an initial homogenizing parameter of graded crushing according to the regulation and control weight; detecting the viscosity of the suspension subjected to primary crushing according to the initial crushing parameters, predicting the rationality of the initial homogenizing parameters according to the viscosity of the suspension, and performing secondary determination on the initial homogenizing parameters; performing homogenizing pressure compensation on the initial homogenizing parameters according to a suspension viscosity driving enzymolysis strategy; detecting impedance of each suspension sample, determining wall breaking degree according to impedance change, and adjusting centrifugal parameters. The germs are subjected to graded crushing, preset parameters are adjusted, and the yield of active ingredients is increased.

Description

Technical Field

[0001] This invention relates to the field of separation and extraction technology, and in particular to a method for extracting plant embryonic stem cells based on high-quality plant embryos. Background Technology

[0002] Plant embryo extract is a complex of active substances extracted from the embryonic tissue during plant seed germination. It is rich in nutrients such as protein, vitamins, polyphenols, trace elements, and minerals. Compared to animal-derived embryo extract, plant embryo extract has advantages such as high safety, lack of ethical controversy, and good biocompatibility, and is widely used in the fields of beauty and skincare, pharmaceuticals, and functional foods. Studies have shown that plant embryo extract can effectively promote cell regeneration, enhance skin protein synthesis, inhibit tyrosinase activity (whitening effect), and has significant antioxidant (DPPH free radical scavenging) and anti-aging effects.

[0003] Currently, the main extraction methods for plant embryo extracts include ultrasound-assisted extraction, membrane separation purification, and low-temperature water extraction. However, the following key issues still exist: traditional methods do not grade the embryos, resulting in the mixed extraction of embryos at different activity stages, which affects the content of active ingredients in the final embryo extract; corn, soybean, and other embryos often have endosperm residues, and the starch in them can adsorb active ingredients (such as polyphenols), reducing the extraction rate and increasing the difficulty of subsequent purification; conventional ultrasound extraction can only break 60-80% of the cell walls, and some active substances remain in the unbroken cells.

[0004] Chinese Patent Publication No. CN117680368A discloses a method for extracting corn embryonic pigments that can effectively improve collection efficiency. The method includes: Step 1, placing standard-compliant corn seeds into a culture chamber; Step 2, determining the proportion of buds in different states within the culture chamber; Step 3, determining the quality of buds in different states; Step 4, transporting the corn buds to a sorting device for sorting and collection; Step 5, washing suitable-quality corn buds with distilled water, adding distilled water, and grinding them in a grinder to obtain a germ mixture; Step 6, ultrasonically extracting the germ mixture, then centrifuging to remove solid impurities, obtaining a corn embryonic pigment filtrate; Step 7, disinfecting the corn embryonic pigment filtrate to obtain a corn embryonic pigment sample solution. This invention reduces manual sorting time by supplementing the sorting device, thereby effectively improving the collection efficiency of corn embryonic pigments. However, the method for extracting corn embryonic pigments that effectively improves collection efficiency has the following problems:

[0005] Traditional plant embryo extract extraction methods suffer from significant variations in plant germ raw materials and rigid extraction processes, making them unsuitable for the precise processing of high-oil-content germs such as corn. Summary of the Invention

[0006] Therefore, this invention provides a method for extracting plant embryonic stem cells based on high-quality plant germs, in order to overcome the problems in the prior art where there are large differences in the raw materials of plant germs, and the increased solution viscosity after crushing high-oil-content germs such as corn affects the extraction accuracy.

[0007] To achieve the above objectives, the present invention provides a method for extracting plant embryogenesis based on high-quality plant embryos, comprising:

[0008] Corn germ was added to a solvent according to the initial solid-liquid ratio, and then graded, crushed, water-extracted, centrifuged, and filtered.

[0009] Germination degree is defined based on changes in corn germ weight and radicle length to determine the germination state of the current batch of corn germ.

[0010] The initial grinding parameters for the graded grinding are determined based on the integrity of the surface waxy layer of the corn germ and the degree of germination.

[0011] The control weights are determined based on the integrity of the surface wax layer and the amount of endosperm residue. The viscosity change of the suspension after primary pulverization according to the initial pulverization parameters is predicted. The initial homogenization parameters for the graded pulverization are determined based on the control weights.

[0012] The viscosity of the suspension after primary pulverization according to the initial pulverization parameters is detected, and the rationality of the initial homogenization parameters is predicted based on the suspension viscosity. The initial homogenization parameters are then determined a second time.

[0013] Based on the suspension viscosity-driven enzymatic hydrolysis strategy, homogenization pressure compensation is applied to the initial homogenization parameters.

[0014] The impedance of each suspension sample was measured, and the degree of cell disruption was determined based on the impedance changes, and the centrifugation parameters were adjusted accordingly.

[0015] Furthermore, the process of determining the germination state includes calculating the degree of germination based on weight changes and radicle length.

[0016] If the actual germination rate is greater than or equal to the preset germination rate, the plant embryo is judged to be in the first germination state, the current batch of plant embryos meets the standard, and plant embryos are added according to the initial solid-liquid ratio;

[0017] If the actual germination rate is less than the preset germination rate, the plant embryo is judged to be in the second germination state, and the current batch of plant embryos is judged to be substandard and replaced with the next batch.

[0018] Furthermore, when the current batch of corn germ is in the first germination stage, the plant germ is graded and crushed, and the initial ultrasonic power in the initial crushing parameters is determined based on the integrity of the surface wax layer and the degree of germination.

[0019] The graded pulverization includes primary pulverization and secondary pulverization. The primary pulverization is ultrasonic extraction pulverization, and the secondary pulverization is high-pressure homogenization pulverization.

[0020] Furthermore, the control weights were determined based on the integrity of the surface wax layer and the amount of residual endosperm;

[0021] When the control weight is less than the control coefficient, the homogenization temperature is reduced according to the ratio of the control weight to the control coefficient, and the homogenization pressure is increased according to the ratio of the control coefficient to the control weight.

[0022] Furthermore, the viscosity of the suspension after primary pulverization was measured;

[0023] When the viscosity of the suspension is detected to be greater than the critical viscosity, it is determined that the viscosity of the suspension has suddenly increased, and the cause of the sudden increase in the viscosity of the suspension is determined based on the amount of endosperm residue.

[0024] Furthermore, when the viscosity of the suspension suddenly increases, it is determined that the initial homogenization parameters need to be adjusted a second time, triggering low-temperature homogenization. Secondary pulverization is then performed according to the adjusted initial homogenization parameters, and the initial solid-liquid ratio of the plant embryo and the solvent is adjusted.

[0025] Furthermore, the process according to the suspension viscosity-driven enzymatic hydrolysis strategy includes,

[0026] When the viscosity of the suspension exceeds the preset viscosity range, cellulase is added; when the viscosity of the suspension is lower than the preset viscosity range, the degree of cell wall disruption of plant embryo cells after graded pulverization is determined.

[0027] Homogenization pressure compensation is performed based on the homogenization pressure of the initial homogenization parameter when the viscosity of the suspension increases, and the homogenization pressure is positively correlated with the viscosity increase.

[0028] Furthermore, the process of determining the cause of the sudden increase in the viscosity of the suspension includes,

[0029] When the residual endosperm content is greater than 5%, the reason for the sudden increase in the viscosity of the suspension is determined to be that the residual endosperm causes starch to dissolve.

[0030] Adjust the initial concentration of vitamin E according to the range of the antioxidant index. When the antioxidant index is within the range, add vitamin E according to the initial concentration.

[0031] When the antioxidant index exceeds the range, increase the initial concentration of vitamin E. When the antioxidant index is below the range, determine that the oxidation sensitivity exceeds the safe range and add an antioxidant.

[0032] Furthermore, the degree of cell wall disruption after graded pulverization is determined based on the impedance change.

[0033] The actual cell wall breakage rate is calculated based on the impedance of each suspension sample. If the actual cell wall breakage rate is greater than or equal to the standard cell wall breakage rate, the degree of cell wall breakage is judged to meet the cell wall breakage requirements.

[0034] If the actual cell wall breakage rate is less than the standard cell wall breakage rate, it is determined that the degree of cell wall breakage does not meet the cell wall breakage requirements, and the centrifugation parameters are adjusted.

[0035] Furthermore, the process of adjusting centrifugation parameters includes,

[0036] When the degree of cell wall disruption does not meet the requirements, the centrifugation time should be increased according to the ratio of the standard cell wall disruption rate to the actual cell wall disruption rate.

[0037] Compared with the prior art, the beneficial effects of the present invention are that the method fully considers the characteristics of corn germ such as high oil content (40-50%), easy oxidation and aleurone layer barrier, solves the bottleneck problem of extraction after corn germ is crushed, and improves the yield of active ingredients in embryo extract compared with the general plant embryo extract method.

[0038] Furthermore, during the extraction of plant embryo extract from plant germ, plant germs exhibit varying weights and germination degrees. This method adheres to the fundamental principle that "high-quality raw materials determine high-quality products," defining the germination degree through changes in corn germ weight and radicle length. This quantifies the germination degree of different batches of plant germs and optimizes the subsequent embryo extract extraction process based on the actual germination degree, solving the industry problem of "large raw material differences and rigid processes" in traditional extraction methods. Since the waxy layer on the corn surface affects the hardness of the germ, and the starch contained in the corn endosperm directly affects the viscosity of the suspension, this method employs a graded pulverization extraction strategy. The initial ultrasonic power in the primary pulverization is determined based on the integrity of the corn germ's surface waxy layer and the germination degree. Further, control weights are defined based on the integrity of the surface waxy layer and the amount of residual endosperm. Homogenization parameters in the secondary pulverization are pre-adjusted according to these control weights, improving the flexibility and adaptability of embryo extract extraction from different batches of corn germ.

[0039] Furthermore, after corn germ is pulverized, its viscosity and high oil content (40-50%) lead to an increase in the viscosity of the pulverized suspension (>200 cP), hindering solid-liquid separation. This method predicts the rationality of the preset parameters for graded pulverization by analyzing the viscosity change of the suspension after primary pulverization. Based on the suspension viscosity, the preset initial homogenization parameters are adjusted a second time. The secondary pulverization is performed by cooling and pressurizing to increase extraction efficiency and reduce the impact of viscosity. Low-temperature homogenization can also prevent thermal damage. The secondary pulverization is performed according to the adjusted initial homogenization parameters, and the initial solid-liquid ratio of plant germ and solvent in the next batch is reduced accordingly. At the same time, the enzymatic hydrolysis strategy is driven by viscosity data to reduce the impact of viscosity changes on the subsequent embryonic extract extraction process. Furthermore, the cause of the sudden increase in suspension viscosity is determined based on the endosperm residue (ER). An antioxidant index is defined based on germination degree, surface wax layer integrity, and endosperm residue. The concentration and type of additives are adjusted according to the defined antioxidant index range.

[0040] Furthermore, during the cell wall disruption process in plant cells, the loss of cell membrane integrity leads to the leakage of intracellular electrolytes, causing regular changes in the dielectric properties of the suspension. This method quantifies the degree of cell wall disruption by measuring impedance changes at specific frequencies. Due to increased viscosity, centrifugation efficiency decreases, membrane filtration flux declines, and ultrafiltration time is prolonged. Therefore, the centrifugation time needs to be adjusted according to the viscosity change value to increase the degree of embryonic extract extraction. At the same time, since temperature affects the impedance of the suspension sample, this method compensates for temperature drift in the cell wall disruption rate, correcting the reading by 0.8% per °C change, eliminating the influence of environmental factors, increasing detection accuracy, and adapting to the closed-loop control requirements of electromechanical coupling cell wall disruption processes through non-destructive, rapid, and quantitative detection methods. Attached Figure Description

[0041] Figure 1 This is a schematic flowchart of the plant embryo extract method based on high-quality plant embryos in an embodiment of the present invention.

[0042] Figure 2 This is a flowchart illustrating the determination of the germination state of corn germ in the current batch, as described in this embodiment of the invention.

[0043] Figure 3 This is a flowchart illustrating the determination process for adjusting the initial concentration based on the antioxidant index range in this embodiment of the invention.

[0044] Figure 4 This is a flowchart illustrating the determination process for whether the degree of cell wall disruption meets the requirements in an embodiment of the present invention. Detailed Implementation

[0045] To make the objectives and advantages of the present invention clearer, the present invention will be further described below with reference to embodiments; it should be understood that the specific embodiments described herein are merely for explaining the present invention and are not intended to limit the present invention.

[0046] Preferred embodiments of the present invention will now be described with reference to the accompanying drawings. Those skilled in the art should understand that these embodiments are merely illustrative of the technical principles of the present invention and are not intended to limit the scope of protection of the present invention.

[0047] It should be noted that in the description of this invention, the terms "upper", "lower", "left", "right", "inner", "outer", etc., which indicate directions or positional relationships, are based on the directions or positional relationships shown in the accompanying drawings. This is only for the convenience of description and is not intended to indicate or imply that the device or element must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, it should not be construed as a limitation of this invention.

[0048] Furthermore, it should be noted that, in the description of this invention, unless otherwise explicitly specified and limited, the terms "installation," "connection," and "linking" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two components. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0049] Please see Figures 1-4 As shown, Figure 1 This is a schematic flowchart of the plant embryo extract method based on high-quality plant embryos in an embodiment of the present invention. Figure 2 This is a flowchart illustrating the determination of the germination state of corn germ in the current batch, as described in this embodiment of the invention. Figure 3 This is a flowchart illustrating the determination process for adjusting the initial concentration based on the antioxidant index range in this embodiment of the invention. Figure 4 This is a flowchart illustrating the determination process for whether the degree of cell wall disruption meets the requirements in an embodiment of the present invention.

[0050] This invention provides a method for extracting plant embryogenesis based on high-quality plant embryos, comprising:

[0051] Step S1: Add corn germ to solvent according to the initial solid-liquid ratio, grade, crush and extract with water, centrifuge and filter.

[0052] Step S2: Define the germination degree based on the changes in corn germ weight and radicle length, and determine the germination state of the current batch of corn germ.

[0053] Step S3: Determine the initial grinding parameters for the graded grinding based on the integrity of the surface wax layer of the corn germ and the degree of germination.

[0054] Step S4: Determine the control weight based on the integrity of the surface wax layer and the amount of endosperm residue, predict the viscosity change of the suspension after primary pulverization according to the initial pulverization parameters, and determine the initial homogenization parameters for the graded pulverization based on the control weight.

[0055] Step S5: Detect the viscosity of the suspension after primary pulverization according to the initial pulverization parameters, and determine the initial homogenization parameters a second time based on the reasonableness of the prediction of the initial homogenization parameters according to the suspension viscosity.

[0056] Step S6: According to the suspension viscosity-driven enzymatic hydrolysis strategy, the initial homogenization parameters are compensated for homogenization pressure.

[0057] Step S7: Detect the impedance of each suspension sample, determine the degree of cell disruption based on the impedance change, and adjust the centrifugation parameters.

[0058] In this embodiment, the plant embryo is a corn embryo.

[0059] Specifically, this method fully considers the characteristics of corn germ, such as its high oil content (40-50%), easy oxidation, and aleurone layer barrier, and solves the bottleneck problem of extraction after corn germ is crushed. Compared with the general plant embryo extract extraction method, it improves the yield of active ingredients in embryo extract.

[0060] Before the crushing process, the surface wax layer integrity (WI) and endosperm residue (ER) of corn germ are tested. Extraction parameters and types of regulators are adjusted, and the initial crushing parameters are determined in combination with the germination degree (GD).

[0061] The initial pulverization parameters for the graded pulverization are determined based on the surface wax layer integrity (WI) and germination degree (GD), and the initial homogenization parameters for the graded pulverization are predicted.

[0062] Determine the germination status of the corn germ in the current batch.

[0063] When the corn germ of the current batch is in the first germination stage, the corn germ is graded and crushed, and the initial ultrasonic power in the initial crushing parameters is determined according to the surface wax layer integrity (WI) and germination degree (GD).

[0064] Specifically, the integrity of the surface wax layer WI of corn germ is detected by near-infrared reflectance, wherein the initial ultrasonic power P = 200 + 100 × GD - 50 × WI.

[0065] In practice, the graded crushing includes primary crushing and secondary crushing of the screened corn germ. The primary crushing uses an ultrasonic extraction device to ultrasonically extract and crush the corn germ, and the secondary crushing uses a high-pressure homogenizer to homogenize and crush the corn germ after the primary crushing.

[0066] The process of determining the germination state includes,

[0067] The weight change of the current batch of plant embryos is detected according to the initial detection cycle. The radicle morphology is captured by a near-infrared camera. The radicle length is detected by image recognition. The germination degree is calculated based on the weight change and radicle length to determine the germination state of the current batch of plant embryos.

[0068] The germination rate calculation formula is: GD = 0.3(ΔW / ΔWm) + 0.4(Lr / Lrm);

[0069] In the formula, GD is the germination degree, ΔW is the weight change range, Lr is the proportion of radicle length, ΔWm is the standard change range, and Lrm is the standard length proportion.

[0070] In this embodiment, ΔWm is 25% and Lrm is 1.6.

[0071] If the actual germination rate is greater than or equal to the preset germination rate, the plant embryo is judged to be in the first germination state, the current batch of plant embryos meets the standard, and plant embryos are added according to the initial solid-liquid ratio;

[0072] If the actual germination rate is less than the preset germination rate, the plant embryo is judged to be in the second germination state, and the current batch of plant embryos is judged to be substandard and replaced with the next batch.

[0073] The first preset germination rate is 0.7.

[0074] Based on the surface wax layer integrity (WI) and endosperm residue (ER), the viscosity change caused by starch and oiliness after high-pressure homogenization after primary grinding is predicted, and the initial homogenization parameters are determined, including the initial homogenization temperature and the initial homogenization pressure.

[0075] In practice, the residual endosperm content refers to the proportion of starchy endosperm tissue remaining on the surface and at the junction of corn germ. The residual endosperm content (ER) of the current batch of corn germ is measured by near-infrared spectroscopy.

[0076] The initial homogenization parameter in the secondary pulverization is adjusted according to the predicted control weight, where the control weight is equal to 0.3×(1-WI)+0.2×ER. When the control weight is less than the control coefficient, the initial homogenization parameter is adjusted.

[0077] Specifically, the homogenization temperature is reduced according to the ratio of the control weight to the control coefficient, and the homogenization pressure is increased according to the ratio of the control coefficient to the control weight.

[0078] The regulation coefficient is 0.31.

[0079] Specifically, in the process of extracting plant embryos from plant germ, the plant germs vary in weight and germination degree. This method follows the basic principle that "high-quality raw materials determine high-quality products." It defines the germination degree by the weight change and root length of the corn germ, quantifies the germination degree of different batches of plant germs, and optimizes the subsequent embryo extraction process based on the actual germination degree, thus solving the industry problem of "large raw material differences and rigid processes" in traditional extraction methods. Furthermore, since the waxy layer on the surface of corn affects the hardness of the germ, and the starch contained in the corn endosperm directly affects the viscosity of the suspension, this method adopts a graded crushing extraction strategy. The initial ultrasonic power in the primary crushing is determined based on the integrity of the surface waxy layer of the corn germ and the germination degree. The control weight is further defined based on the integrity of the surface waxy layer and the amount of residual endosperm. The homogenization parameters in the secondary crushing are pre-adjusted according to the control weight, which improves the flexibility and adaptability of embryo extraction from different batches of corn germ.

[0080] A viscosity sensor is used to detect the viscosity of the suspension, and the rationality of the staged grinding parameters is predicted and judged based on the viscosity change of the suspension after primary grinding.

[0081] When the viscosity of the suspension is detected to be greater than the critical viscosity, it is determined that the viscosity of the suspension has suddenly increased, and the cause of the sudden increase in the viscosity of the suspension is determined.

[0082] When the viscosity of the suspension suddenly increases, it is determined that the initial homogenization parameters after the prediction adjustment need to be adjusted a second time, triggering low-temperature homogenization, performing secondary pulverization according to the initial homogenization parameters after the second adjustment, and adjusting the initial solid-liquid ratio of the plant embryos and solvent added in the next batch.

[0083] Specifically, the low-temperature homogenization involves lowering the initial homogenization temperature to 4°C and reducing the initial solid-liquid ratio based on the ratio of the critical viscosity to the suspension viscosity.

[0084] The critical viscosity is 150 cP.

[0085] Based on the enzymatic hydrolysis strategy driven by online viscosity sensor data, cellulase is added when the viscosity of the suspension exceeds the preset viscosity range, and the degree of cell wall disruption of plant embryo cells after grading and pulverization is directly determined when the viscosity of the suspension is lower than the preset viscosity range.

[0086] During implementation, the homogenization pressure of the high-pressure homogenizer is increased according to the viscosity change to compensate for the high-pressure homogenization pressure. The homogenization pressure is positively correlated with the viscosity increase rate. For every 1% increase in viscosity increase rate, the homogenization pressure increases by 2 MPa.

[0087] The amount of cellulase added is 0.5 U / g, and the preset viscosity range is 80 cP to 180 cP.

[0088] The cause of the sudden increase in the viscosity of the suspension was determined based on the residual endosperm ER. When the residual endosperm ER was greater than 5%, the cause of the sudden increase in the viscosity of the suspension was determined to be starch dissolution caused by high ER.

[0089] After determining the cause, adjust the initial concentration of vitamin E according to the range of the antioxidant index. When the antioxidant index is within the range, add vitamin E according to the initial concentration.

[0090] Antioxidant index = (1-GD)×0.6 + WI×0.2 + ER×0.2;

[0091] When the antioxidant index exceeds the range, increase the initial concentration of vitamin E.

[0092] When the antioxidant index is below the specified range, and the oxidation sensitivity is determined to be outside the safe range, an antioxidant is added.

[0093] In this embodiment, the antioxidant is glutathione.

[0094] The index ranges from 0.5 to 0.8, and the initial concentration is 120 mg / L.

[0095] Specifically, after corn germ is pulverized, its high viscosity and oil content (40-50%) lead to an increase in the viscosity of the pulverized suspension (>200 cP), hindering solid-liquid separation. This method predicts the rationality of the preset parameters for graded pulverization by analyzing the viscosity change of the suspension after primary pulverization. Based on the suspension viscosity, the preset initial homogenization parameters are adjusted a second time. The secondary pulverization is performed by cooling and pressurizing to increase extraction efficiency and reduce the impact of viscosity. Low-temperature homogenization can also prevent thermal damage. The secondary pulverization is performed according to the adjusted initial homogenization parameters, and the initial solid-liquid ratio of plant germ and solvent in the next batch is reduced accordingly. At the same time, the enzymatic hydrolysis strategy is driven by viscosity data to reduce the impact of viscosity changes on the subsequent embryonic extract extraction process. Furthermore, the cause of the sudden increase in suspension viscosity is determined based on the endosperm residue (ER). An antioxidant index is defined based on germination degree, surface wax layer integrity, and endosperm residue. The concentration and type of additives are adjusted according to the defined antioxidant index range.

[0096] During the extraction of embryonic material, suspension samples were extracted, and impedance analyzers were used to detect the impedance of each suspension sample. The degree of cell wall disruption was determined based on the impedance changes.

[0097]

[0098] In the formula, Z2 is the impedance of the suspension sample before cell disruption, Z1 is the impedance of the suspension sample after cell disruption, Z0 is the predicted impedance of the completely disrupted suspension, and ΔT is the temperature change between the suspension sample and the completely disrupted suspension.

[0099] If the actual cell wall breakage rate is greater than or equal to the standard cell wall breakage rate, then the degree of cell wall breakage is judged to meet the cell wall breakage requirements.

[0100] If the actual cell wall breakage rate is less than the standard cell wall breakage rate, it is determined that the degree of cell wall breakage does not meet the cell wall breakage requirements, and the centrifugation parameters are adjusted.

[0101] Specifically, the centrifugation time is increased based on the ratio of the standard cell breakage rate to the actual cell breakage rate;

[0102] The standard cell wall breakage rate is 90%, and the temperature compensation coefficient is 0.8%.

[0103] Specifically, during the cell wall disruption process in plant cells, the loss of cell membrane integrity leads to the leakage of intracellular electrolytes, causing regular changes in the dielectric properties of the suspension. This method quantifies the degree of cell wall disruption by measuring impedance changes at specific frequencies. Due to increased viscosity, centrifugation efficiency decreases, membrane filtration flux declines, and ultrafiltration time is prolonged. Therefore, the centrifugation time needs to be adjusted according to the viscosity change value to increase the degree of embryonic extract extraction. At the same time, since temperature affects the impedance of the suspension sample, this method compensates for temperature drift in the cell wall disruption rate, correcting the reading by 0.8% per °C change, eliminating the influence of environmental factors, increasing detection accuracy, and adapting to the closed-loop control requirements of electromechanical coupling cell wall disruption processes through non-destructive, rapid, and quantitative detection methods.

[0104] The technical solution of the present invention has been described above with reference to the preferred embodiments shown in the accompanying drawings. However, it will be readily understood by those skilled in the art that the scope of protection of the present invention is obviously not limited to these specific embodiments. Without departing from the principles of the present invention, those skilled in the art can make equivalent changes or substitutions to the relevant technical features, and the technical solutions after these changes or substitutions will all fall within the scope of protection of the present invention.

[0105] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for extracting plant embryonic stem cells based on high-quality plant embryos, characterized in that, include: Corn germ was added to a solvent according to the initial solid-liquid ratio, and then graded, crushed, water-extracted, centrifuged, and filtered. Germination degree is defined based on changes in corn germ weight and radicle length to determine the germination state of the current batch of corn germ. The initial grinding parameters for the graded grinding are determined based on the integrity of the surface waxy layer of the corn germ and the degree of germination. The control weights are determined based on the integrity of the surface wax layer and the amount of endosperm residue. The viscosity change of the suspension after primary pulverization according to the initial pulverization parameters is predicted. The initial homogenization parameters for the graded pulverization are determined based on the control weights. The viscosity of the suspension after primary pulverization according to the initial pulverization parameters is detected, and the rationality of the initial homogenization parameters is predicted based on the suspension viscosity. The initial homogenization parameters are then determined a second time. Based on the suspension viscosity-driven enzymatic hydrolysis strategy, homogenization pressure compensation is applied to the initial homogenization parameters. The impedance of each suspension sample was measured, and the degree of cell disruption was determined based on the impedance changes, and the centrifugation parameters were adjusted accordingly.

2. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 1, characterized in that, The process of determining the germination state includes calculating the degree of germination based on weight changes and radicle length. If the actual germination rate is greater than or equal to the preset germination rate, the plant embryo is judged to be in the first germination state, the current batch of plant embryos meets the standard, and plant embryos are added according to the initial solid-liquid ratio; If the actual germination rate is less than the preset germination rate, the plant embryo is judged to be in the second germination state, and the current batch of plant embryos is judged to be substandard and replaced with the next batch.

3. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 2, characterized in that, When the current batch of corn germ is in the first germination stage, the plant germ is graded and crushed, and the initial ultrasonic power in the initial crushing parameters is determined based on the integrity of the surface wax layer and the degree of germination. The graded pulverization includes primary pulverization and secondary pulverization. The primary pulverization is ultrasonic extraction pulverization, and the secondary pulverization is high-pressure homogenization pulverization.

4. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 3, characterized in that, The control weights are determined based on the integrity of the surface wax layer and the amount of residual endosperm. When the control weight is less than the control coefficient, the homogenization temperature is reduced according to the ratio of the control weight to the control coefficient, and the homogenization pressure is increased according to the ratio of the control coefficient to the control weight.

5. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 4, characterized in that, Detect the viscosity of the suspension after primary grinding; When the viscosity of the suspension is detected to be greater than the critical viscosity, it is determined that the viscosity of the suspension has suddenly increased, and the cause of the sudden increase in the viscosity of the suspension is determined based on the amount of endosperm residue.

6. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 5, characterized in that, When the viscosity of the suspension suddenly increases, it is determined that the initial homogenization parameters need to be adjusted a second time, triggering low-temperature homogenization. Secondary pulverization is then performed according to the adjusted initial homogenization parameters, and the initial solid-liquid ratio of the plant embryo and the solvent is adjusted.

7. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 6, characterized in that, The process according to the suspension viscosity-driven enzymatic hydrolysis strategy includes: When the viscosity of the suspension exceeds the preset viscosity range, cellulase is added; when the viscosity of the suspension is lower than the preset viscosity range, the degree of cell wall disruption of plant embryo cells after graded pulverization is determined. Homogenization pressure compensation is performed based on the homogenization pressure of the initial homogenization parameter when the viscosity of the suspension increases, and the homogenization pressure is positively correlated with the viscosity increase.

8. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 7, characterized in that, The process of determining the cause of the sudden increase in the viscosity of the suspension includes, When the residual endosperm content is greater than 5%, the reason for the sudden increase in the viscosity of the suspension is determined to be that the residual endosperm causes starch to dissolve. Adjust the initial concentration of vitamin E according to the range of the antioxidant index. When the antioxidant index is within the range, add vitamin E according to the initial concentration. When the antioxidant index exceeds the range, increase the initial concentration of vitamin E. When the antioxidant index is below the range, determine that the oxidation sensitivity exceeds the safe range and add an antioxidant.

9. The method for extracting plant embryonic stem cells based on high-quality plant embryos according to claim 8, characterized in that, The degree of cell wall breakage after graded pulverization is determined based on impedance changes. The actual cell wall breakage rate is calculated based on the impedance of each suspension sample. If the actual cell wall breakage rate is greater than or equal to the standard cell wall breakage rate, the degree of cell wall breakage is judged to meet the cell wall breakage requirements. If the actual cell wall breakage rate is less than the standard cell wall breakage rate, it is determined that the degree of cell wall breakage does not meet the cell wall breakage requirements, and the centrifugation parameters are adjusted.

10. The method for extracting plant embryogenesis based on high-quality plant embryos according to claim 9, characterized in that, The process of adjusting centrifugation parameters includes, When the degree of cell wall disruption does not meet the requirements, the centrifugation time should be increased according to the ratio of the standard cell wall disruption rate to the actual cell wall disruption rate.

Images