CN117462577A - Method for extracting total flavonoids from elm pod - Google Patents

Abstract

The invention discloses a method for extracting total flavonoids from elm seeds, and relates to the technical field of flavone extraction. The invention comprises the following steps: s1: material preparation: a dried elm pod sample; ethanol or ethanol-water mixed solvent; s2: crushing a sample: grinding the dried elm pod sample to a fine powder; s3: extracting: sample preparation: the elm pod samples were chopped or ground into smaller particles to increase surface area, aid in extraction, ensure sample drying to avoid the effect of moisture on extraction. The invention is prepared by the following materials: preparing a dried elm pod sample, ensuring the quality and purity of the sample, preparing ethanol or ethanol-water mixed solvent, crushing the sample for extracting total flavonoids in the elm pod, grinding the dried elm pod sample into fine powder to increase the surface area of the sample, facilitating the extraction of flavonoid compounds, and mixing the crushed elm pod sample with ethanol according to a certain proportion.

Description

Method for extracting total flavonoids from elm pod

Technical Field

The invention belongs to the technical field of flavone extraction, and particularly relates to a method for extracting total flavonoids from elm seeds.

Background

Elm pod seed, the name of the traditional Chinese medicine. Is fruit or seed of Ulmus pumila of Ulmaceae. Distributed in northeast, north China, east China, south China, southwest, tibetan etc., and cultivated in multiple lines in the south of Yangtze river. Has effects of invigorating spleen, tranquilizing, clearing heat, promoting diuresis, relieving swelling and killing parasite. It is commonly used for insomnia, inappetence, leukorrhagia, dysuria, edema, infantile malnutrition, emaciation, burn, scald, and tinea. Elm, also known as green, white elm, green elm, qian Shu, elm, hammer, elm, and elm. The largeleaf arbor grows into shrubs in dry barren lands up to 25 meters and 1 meter in chest diameter; the bark of the sapling is smooth, grey brown or light grey, the bark of the big tree is dark grey, irregular deep and longitudinal crack and rough; the branches are hairless or have hair, light yellow gray, light brown gray or gray, light brown yellow or yellow, have scattered skin holes, have no expanded wood plug layers and raised wood plug wings; the winter bud is nearly spherical or oval, no hair is arranged on the back of the bud scale, and the edge of the bud scale on the inner layer is provided with white long soft hair. She Tuoyuan oval, long oval, elliptic needle-like or oval needle-like, 2-8 cm long and 1.2-3.5 cm wide, the tip is pointed or pointed, the base is oblique or nearly symmetrical, one side is wedge-shaped to round, the other side is round to half heart-shaped, the leaf surface is smooth and hairless, the leaf back is short and soft, after that, hairless or partial axilla has tufted hair, the edge has heavy saw teeth or single saw teeth, 9-16 side pulses are on each side, the petiole is 4-10 mm long, and usually only short and soft hair is on the upper surface. Flowers have first leaf opening and cluster in the axilla of the last year branch. The samara is nearly round, thin and inverted oval and 1.2-2 cm long, except the top gap, the head and the surface of the column are fluffy, the rest is fluffy, the pit part is positioned in the middle of the samara, the upper end is not close to or near the gap, the color of the pit part is the same as that of the fruit wing before and after ripening, the pit part is light green, the pit part is white yellow after ripening, the lodging flower is fluffy, 4 shallow cracks are formed at the edge of the split, the stem is short compared with the flower, the length is 1-2 mm, and the quilt (or thin and non-soft) is short and soft. Flower and fruit period 3-6 months (late northeast).

Although the modern flavone extraction has made great progress, a set of flow system is lacking to sample the flavone, so that in the flavone extraction and sampling process, the quality of the extracted flavone is insufficient due to insufficient unification of sampling methods, and the flavone is to be improved.

Disclosure of Invention

The invention aims to provide a method for extracting total flavonoids from elm seeds, which comprises the following steps of: preparing a dried elm pod sample, ensuring the quality and purity of the sample, preparing ethanol or ethanol-water mixed solvent, crushing the sample for extracting total flavonoids in the elm pod, and grinding the dried elm pod sample into fine powder to increase the surface area of the sample, thereby solving the existing problems.

In order to solve the technical problems, the invention is realized by the following technical scheme:

the invention relates to a method for extracting total flavonoids from elm seeds, which comprises the following steps:

s1: mortar and pestle; filter paper or filter membrane; vacuum concentration equipment; an evaporation dish; a precision balance; a nitrogen purging device;

s2: the extraction efficiency is improved;

s3: ultrasonic auxiliary extraction: the ultrasonic equipment is used for accelerating the extraction process, so that the release of solutes from a sample is improved, and the extraction time is shortened; extracting for multiple times: taking multiple extractions into consideration to improve the yield of total flavonoids, and combining and concentrating the extracting solutions after each extraction; using heat: slightly heating the extract to accelerate the extraction of flavone from the solvent;

s4: can be adjusted according to the requirement, and the longer leaching time is beneficial to extracting more compounds;

s5: obtaining elm pod seed extract;

s6: the nitrogen purging device can be used for assisting in concentration, so that the solvent is removed more thoroughly;

s7: removing residual solvent by evaporation to obtain flavonoid extract;

s8: ensuring the purity and content of the extract;

s9: such as High Performance Liquid Chromatography (HPLC), to determine the total flavone content of the extract.

Preferably, S1 is further expressed as:

s1.1: fresh elm pods typically contain more active ingredient;

s1.2: or mechanically grinding by using a stirrer or other equipment;

s1.3: this can be achieved by screening with a screen or grading the sample using a scale;

s1.4: the storage conditions should be kept dry, cool and the next extraction process should be performed as soon as possible.

Preferably, S2 is further expressed as:

s2.1: ensuring that the sample is not subject to moisture and contamination;

s2.2: and reduces the heat-sensitive damage that may be caused to the active ingredient, precooling typically takes one to two hours;

s2.3: the selection of the appropriate equipment depends on the number and texture of the samples;

s2.4: this helps to improve the effectiveness of the subsequent grinding process;

s2.5: the weight of the sample can be increased appropriately without overcrowding the mortar so as not to affect the polishing effect;

s2.6: until the elm pod samples are completely crushed into fine powder, and when mechanical equipment is used for grinding, proper time and speed are set to ensure that the samples are sufficiently ground;

s2.7: the size and morphology of the sample particles can be observed by using a microscope, and the sample can be classified by using a screening method or the like.

Preferably, S3 is further expressed as:

s3.1: to extract the total flavonoids from elm pods, ethanol is generally a good choice because it is of sufficient polarity to dissolve the flavonoids;

s3.2: typically in a ratio of 1:10 to 1:20, ensuring complete coverage of the elm pod sample in the solvent;

s3.3: the extraction time is usually 12 to 48 hours, and can be adjusted according to the needs, and the longer extraction time is beneficial to extracting more compounds;

s3.4: this step is used to separate the elm pod solid residue from the extract;

s3.5: this helps to reduce the volume of the extract, making subsequent processing more convenient;

s3.6: the evaporation should be carried out at a suitable temperature to avoid damage to the active ingredient by excessive temperatures;

s3.7: this helps to determine the quality and purity of the extracted flavonoids.

Preferably, S4 is further expressed as:

s4.1: the quality and dryness of the sample are ensured, so that the extraction efficiency is improved;

s4.2: common solvents include ethanol, methanol or ethers, and the solvents are selected in consideration of the polarity and solubility of the flavonoids;

s4.3: typically, a ratio of 1:10 to 1:20 is a common option to ensure complete coverage of the elm pod sample in solvent for adequate extraction of the compound of interest;

s4.4: the specific time can be adjusted according to experimental requirements and the characteristics of the elm pod samples, and longer leaching times generally help to extract more compounds;

s4.5: excessive temperatures may cause decomposition or oxidation of the target compound, and thus it is necessary to avoid excessive temperatures;

s4.6: this contributes to an improvement in extraction efficiency;

s4.7: this step is used to separate the elm pod solid residue from the extract;

s4.8: this helps to reduce the volume of the extract, making subsequent processing more convenient.

Preferably, S5 is further expressed as:

s5.1: common devices include filter papers, filter membranes, glass fiber filter papers or other filter materials with appropriate micropore sizes;

s5.2: ensuring the equipment is clean and sterile to avoid contaminating the extract;

s5.3: this helps to improve the filtration efficiency;

s5.4: if clogging occurs, the filter paper can be replaced or the filtrate can be carried out, and sometimes it is helpful to pre-filter the extract to remove large-particle solids;

s5.5: ensuring the container clean so as not to pollute the extract;

s5.6: such as repeated concentration, crystallization, etc., to obtain purer elm pod total flavone extract;

s5.7: for the next use.

Preferably, S6 is further expressed as:

s6.1: ensure that it is clean and sterile when needed to avoid contamination;

s6.2: adding an extracting solution: pouring the elm pod total flavone extracting solution into an evaporation container;

s6.3: typically between 40 ℃ and 60 ℃, excessive temperatures may lead to degradation of the flavonoids, if it is desired to speed up the concentration process, it is conceivable to use a vacuum pump to reduce the ambient pressure in order to reduce the boiling point of the solvent and accelerate evaporation;

s6.4: to ensure a uniform concentration process which helps to avoid the formation of crystals or scale;

s6.5: this may take a period of time depending on the initial extract concentration and heating conditions;

s6.6: allowing the liquid in the container to cool naturally to room temperature;

s6.7: or nitrogen is used for protecting the liquid level in the concentration process;

s6.8: the final transfer can be performed using a solvent if desired;

s6.9: such as high performance liquid chromatography or mass spectrometry, to determine the exact flavone content of the concentrate;

s6.10: typically refrigerated or frozen, to prevent degradation.

Preferably, S7 is further expressed as:

s7.1: cleaning and fine crushing, and removing impurities and dust by using a castor brush;

s7.2: this may be done at room temperature or with slight heating to facilitate extraction;

s7.3: to ensure adequate extraction of the flavonoids;

s7.4: collecting filtrate, which is the extractive solution containing total flavonoids;

s7.5: if the evaporation process needs to be accelerated, a vacuum pump can be considered to reduce the ambient pressure so as to reduce the boiling point of the solvent and accelerate the evaporation;

s7.6: this helps to avoid the formation of crystals or scale.

Preferably, S8 is further expressed as:

s8.1: this will construct a standard curve for the subsequent calculation of total flavone content;

s8.2: until the volume of the extract is reduced to the desired concentration;

s8.3: quantification was performed with a standard curve;

s8.4: calculating the total flavone content in the elm pod sample;

s8.5: meanwhile, the measurement result of the total flavone content is reported, including the standard curve and the correction step used.

Preferably, S8 is further expressed as:

s9.1: solvents of high purity, such as methanol or ethanol, are typically used;

s9.2: setting appropriate HPLC conditions including flow rate, column temperature, detection wavelength, etc.; injecting a sample, running analysis, and recording the peak area of the flavonoid compound;

s9.3: calculating the concentration of flavonoid compounds in the sample according to a standard curve;

s9.4: calculating the content of total flavonoids in elm pod kernels, and reporting the result;

s9.5: including repeated analyses using quality control samples and standards.

The invention has the following beneficial effects:

1. the invention is prepared by the following materials: preparing a dried elm pod sample, ensuring the quality and purity of the sample, preparing ethanol or ethanol-water mixed solvent, crushing the sample for extracting total flavonoids in the elm pod, grinding the dried elm pod sample into fine powder to increase the surface area of the sample, facilitating the extraction of flavonoid compounds, and mixing the crushed elm pod sample with ethanol according to a certain proportion.

2. The invention is sealed by the mixture: placing the mixture of ethanol and elm pod sample in a sealed container, wherein the sealing is used for preventing external air from entering, so as to reduce the oxidation influence of oxygen in the sample on the flavone compounds; leaching at room temperature: the extraction is carried out by placing the sealed vessel at room temperature, which operating conditions help to maintain the stability of the flavonoid compounds, while also facilitating laboratory operations, and which may be advantageous for extraction of certain specific compounds while avoiding sample degradation that may result from high temperatures.

3. The invention prepares the filter: placing the funnel on a collection container, and placing filter paper or a filter membrane in the funnel to ensure that the filter paper completely covers the hole of the funnel; wetting filter paper: wetting the filter paper with a small amount of extraction solution, which helps the filter paper to adhere tightly to the funnel and prevents liquid from flowing around the filter paper; pouring the mixture: the mixture containing the solid particles was slowly poured into a funnel and the liquid was filtered through a filter paper while the solid particles were left on the filter paper.

Of course, it is not necessary for any one product to practice the invention to achieve all of the advantages set forth above at the same time.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed for the description of the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a flow chart of the present invention.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

Referring to fig. 1, the invention relates to a method for extracting total flavonoids from elm seeds, which comprises the following steps:

s1: mortar and pestle; filter paper or filter membrane; vacuum concentration equipment; an evaporation dish; a precision balance; a nitrogen purging device;

s2: the extraction efficiency is improved;

s3: ultrasonic auxiliary extraction: the ultrasonic equipment is used for accelerating the extraction process, so that the release of solutes from a sample is improved, and the extraction time is shortened; extracting for multiple times: taking multiple extractions into consideration to improve the yield of total flavonoids, and combining and concentrating the extracting solutions after each extraction; using heat: slightly heating the extract to accelerate the extraction of flavone from the solvent;

s4: can be adjusted according to the requirement, and the longer leaching time is beneficial to extracting more compounds;

s5: obtaining elm pod seed extract;

s6: the nitrogen purging device can be used for assisting in concentration, so that the solvent is removed more thoroughly;

s7: removing residual solvent by evaporation to obtain flavonoid extract;

s8: ensuring the purity and content of the extract.

S1 is further expressed as:

s1.1: fresh elm pods typically contain more active ingredient;

s1.2: or mechanically grinding by using a stirrer or other equipment;

s1.3: this can be achieved by screening with a screen or grading the sample using a scale;

s1.4: the storage conditions should be kept dry, cool and the next extraction process should be performed as soon as possible.

Ensuring that the correct elm pod species are selected. Different types of elms may produce different types and levels of flavone compounds. The purpose of the study was clearly studied in selecting samples to determine the type of elm pod needed.

Elm pod sample selection: acquisition time: the flavone content of elm pods may vary with season and growth stage. Generally, the optimal collection time is when the elm pod is mature but not completely cracked, and this is usually in autumn or late summer, a suitable geographical location is selected to collect the elm pod sample, and the soil and climate conditions in different areas may affect the content and type of flavone compounds in the elm pod, so the exact place where the sample is collected should be recorded in the study, and the collected sample should be processed or stored under suitable conditions as soon as possible to prevent degradation of the sample, and typically, the sample may be refrigerated or frozen.

S2 is further expressed as:

s2.1: ensuring that the sample is not subject to moisture and contamination;

s2.2: and reduces the heat-sensitive damage that may be caused to the active ingredient, precooling typically takes one to two hours;

s2.3: the selection of the appropriate equipment depends on the number and texture of the samples;

s2.4: this helps to improve the effectiveness of the subsequent grinding process;

s2.5: the weight of the sample can be increased appropriately without overcrowding the mortar so as not to affect the polishing effect;

s2.6: until the elm pod samples are completely crushed into fine powder, and when mechanical equipment is used for grinding, proper time and speed are set to ensure that the samples are sufficiently ground;

s2.7: the size and morphology of the sample particles can be observed by using a microscope, and the sample can be classified by using a screening method or the like.

Sample selection: ensuring that the elm pod selected is from the correct elm species. Common elm pods may be from multiple varieties of elm genus, each variety may have different composition and efficacy, knowing that the location and season of elm pod collection may have an impact on its chemical composition and quality, e.g., elm pods collected from organic and healthy growing elms may be more desirable, elm pods with complete appearance, no damage and no decay may be selected. Elm pod with vivid color and complete appearance may be more suitable for extraction.

S3 is further expressed as:

s3.1: to extract the total flavonoids from elm pods, ethanol is generally a good choice because it is of sufficient polarity to dissolve the flavonoids;

s3.2: typically in a ratio of 1:10 to 1:20, ensuring complete coverage of the elm pod sample in the solvent;

s3.3: the extraction time is usually 12 to 48 hours, and can be adjusted according to the needs, and the longer extraction time is beneficial to extracting more compounds;

s3.4: this step is used to separate the elm pod solid residue from the extract;

s3.5: this helps to reduce the volume of the extract, making subsequent processing more convenient;

s3.6: the evaporation should be carried out at a suitable temperature to avoid damage to the active ingredient by excessive temperatures;

s3.7: this helps to determine the quality and purity of the extracted flavonoids.

Selecting a suitable solvent: first, it is necessary to understand the nature of the target compound contained in elm pod. In this case, you want to extract the total flavonoids, knowing the chemical nature of these compounds, such as polarity, solubility, can help you choose the proper solvents, the total flavonoids in elm pods are usually polar compounds, so you need to choose a solvent with higher polarity in order to extract these compounds effectively, usually polar solvents such as ethanol, methanol or ethyl acetate are good choices, ensure that the chosen solvents can dissolve the total flavonoids in elm pods effectively, you can perform small-scale experiments, mix elm pod samples with different solvents, and then observe which solvents can dissolve the target compounds better.

S4 is further expressed as:

s4.1: the quality and dryness of the sample are ensured, so that the extraction efficiency is improved;

s4.2: common solvents include ethanol, methanol or ethers, and the solvents are selected in consideration of the polarity and solubility of the flavonoids;

s4.3: typically, a ratio of 1:10 to 1:20 is a common option to ensure complete coverage of the elm pod sample in solvent for adequate extraction of the compound of interest;

s4.4: the specific time can be adjusted according to experimental requirements and the characteristics of the elm pod samples, and longer leaching times generally help to extract more compounds;

s4.5: excessive temperatures may cause decomposition or oxidation of the target compound, and thus it is necessary to avoid excessive temperatures;

s4.6: this contributes to an improvement in extraction efficiency;

s4.7: this step is used to separate the elm pod solid residue from the extract;

s4.8: this helps to reduce the volume of the extract, making subsequent processing more convenient.

Preparing elm pod samples: first, fresh elm pods need to be collected, the elm pods generally mature in autumn, when the elm pods begin to brown, ensuring that elm pods are selected that do not fail, rot or otherwise become apparent, cleaning the elm pods to remove external impurities, gently rinsing the elm pods with warm water, ensuring removal of dirt and other grime, prior to preparation of the elm pod kernel. A soft brush can be used to remove the hard-to-clean dirt, once the elm pod is clean, it is necessary to peel the elm pod to obtain the elm pod kernel, and a knife or finger can be used to gently peel the elm pod shell, ensuring that the kernel inside is not damaged, as complete sample as possible is desired.

S5 is further expressed as:

s5.1: common devices include filter papers, filter membranes, glass fiber filter papers or other filter materials with appropriate micropore sizes;

s5.2: ensuring the equipment is clean and sterile to avoid contaminating the extract;

s5.3: this helps to improve the filtration efficiency;

s5.4: if clogging occurs, the filter paper can be replaced or the filtrate can be carried out, and sometimes it is helpful to pre-filter the extract to remove large-particle solids;

s5.5: ensuring the container clean so as not to pollute the extract;

s5.6: such as repeated concentration, crystallization, etc., to obtain purer elm pod total flavone extract;

s5.7: for the next use.

Selecting an appropriate filtering device: first, it is clear what the substance to be filtered is, whether it is solid, liquid or gas in nature, which can help you determine the type of filtering device that is needed, determine the size range of particle sizes that need to be filtered, some that are more suitable for handling larger particles, while others are suitable for handling tiny particles, determine the flow that is needed, how much substance needs to be handled per unit time, which will affect the size and capacity of the filtering device that you choose.

S6 is further expressed as:

s6.1: ensure that it is clean and sterile when needed to avoid contamination;

s6.2: adding an extracting solution: pouring the elm pod total flavone extracting solution into an evaporation container;

s6.3: typically between 40 ℃ and 60 ℃, excessive temperatures may lead to degradation of the flavonoids, if it is desired to speed up the concentration process, it is conceivable to use a vacuum pump to reduce the ambient pressure in order to reduce the boiling point of the solvent and accelerate evaporation;

s6.4: to ensure a uniform concentration process which helps to avoid the formation of crystals or scale;

s6.5: this may take a period of time depending on the initial extract concentration and heating conditions;

s6.6: allowing the liquid in the container to cool naturally to room temperature;

s6.7: or nitrogen is used for protecting the liquid level in the concentration process;

s6.8: the final transfer can be performed using a solvent if desired;

s6.9: such as high performance liquid chromatography or mass spectrometry, to determine the exact flavone content of the concentrate;

s6.10: typically refrigerated or frozen, to prevent degradation.

Preparing a concentration container: the choice of a suitable concentration vessel, usually a flask, beaker or conical flask, should be made in view of chemistry, volume and ease of handling, to ensure that there are sufficient laboratory equipment such as measuring cups, pipettes, filter papers, glass rods, stirring rods, etc., which should also be chosen to meet the requirements of the experiment, to ensure that the vessel is clean before use, to be rinsed with deionized water or other suitable solvent if the vessel surface is fouled, and to prepare the solution to be concentrated according to the experimental requirements. Ensuring that the concentration and the volume of the solution meet the experimental design requirements.

S7 is further expressed as:

s7.1: cleaning and fine crushing, and removing impurities and dust by using a castor brush;

s7.2: this may be done at room temperature or with slight heating to facilitate extraction;

s7.3: to ensure adequate extraction of the flavonoids;

s7.4: collecting filtrate, which is the extractive solution containing total flavonoids;

s7.5: if the evaporation process needs to be accelerated, a vacuum pump can be considered to reduce the ambient pressure so as to reduce the boiling point of the solvent and accelerate the evaporation;

s7.6: this helps to avoid the formation of crystals or scale.

Sample preparation: ensuring that the target and sample type of the assay are well defined before starting will help to determine the proper sample preparation method, knowing the potential hazards and necessary safety measures, which may include wearing gloves, goggles etc. if the sample needs to be sampled from a large sample, ensuring randomness and representativeness of the sampling effort, sampling using appropriate tools and techniques, and well-defined identification of each sample, including sample number, date and other relevant information, helps to prevent sample confusion.

S8 is further expressed as:

s8.1: this will construct a standard curve for the subsequent calculation of total flavone content;

s8.2: until the volume of the extract is reduced to the desired concentration;

s8.3: quantification was performed with a standard curve;

s8.4: calculating the total flavone content in the elm pod sample;

s8.5: meanwhile, the measurement result of the total flavone content is reported, including the standard curve and the correction step used.

Calibration standard: firstly, proper standard substances are required to be selected according to the requirements of experiments or production, so that the characteristics of elm pods are matched with the physical quantity to be tested, and the used calibration equipment is calibrated and is in a good working state. Furthermore, it is ensured that the calibration device has the same measuring range as the instrument being calibrated, and that the calibration operation is performed in strict steps according to established operating regulations. This includes taking measurements over a series of known values, recording the measurements, and comparing with the known values of the standard, the data collected during calibration needs to be processed and analyzed, which may include calculating errors, determining a calibration curve, or formulating a calibration table, etc.

S9 is further expressed as:

s9.1: solvents of high purity, such as methanol or ethanol, are typically used;

s9.2: setting appropriate HPLC conditions including flow rate, column temperature, detection wavelength, etc.; injecting a sample, running analysis, and recording the peak area of the flavonoid compound;

s9.3: calculating the concentration of flavonoid compounds in the sample according to a standard curve;

s9.4: calculating the content of total flavonoids in elm pod kernels, and reporting the result;

s9.5: including repeated analyses using quality control samples and standards.

Analysis: this step is a general extraction and analysis method, and the specific operation may be adjusted according to the experimental purposes, equipment and materials. Meanwhile, in the extraction and analysis experiments, laboratory safety regulations and regulations must be followed to ensure the safety of the operation.

In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.

The preferred embodiments of the invention disclosed above are intended only to assist in the explanation of the invention. The preferred embodiments are not exhaustive or to limit the invention to the precise form disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best understand and utilize the invention. The invention is limited only by the claims and the full scope and equivalents thereof.

Claims (10)

1. The method for extracting the total flavonoids from the elm seeds is characterized by comprising the following steps: the method comprises the following steps:

s1: material preparation: a dried elm pod sample; ethanol or ethanol-water mixed solvent;

s2: crushing a sample: grinding the dried elm pod sample to a fine powder;

s3: extracting: sample preparation: cutting or grinding the elm pod sample into smaller particles to increase surface area, facilitate extraction, ensure sample drying, and avoid the influence of moisture on extraction; selecting a suitable solvent: selecting the organic solvent most suitable for total flavonoids of elm pod, such as ethanol, methanol or ethyl acetate, wherein the selection of different solvents may affect the extraction efficiency, so that pre-experiments should be performed to determine the optimal solvent; extraction time and temperature: optimizing extraction time and temperature conditions, and improving extraction efficiency of flavone by changing soaking time and temperature, wherein long-time low-temperature soaking is beneficial to improving extraction efficiency;

s4: leaching: sealing the mixture, and leaching at room temperature for 12-48 hr;

s5: and (3) filtering: filtering the extract with filter paper or filter membrane to remove solid particles;

s6: concentrating: concentrating the solvent in the extracting solution by using vacuum concentration equipment to obtain a concentrated extracting solution;

s7: and (3) evaporation: placing the concentrated extract into an evaporation dish, and under a safe condition;

s8: and (3) quality control: quality control of the extract can be performed using chromatographic analysis methods;

s9: analysis: using a suitable analytical instrument.

2. The method of extracting total flavonoids from elm seeds according to claim 1, wherein S1 is further expressed as:

s1.1: elm pod sample selection: selecting fresh and dried elm pod as the raw material for extraction;

s1.2: sample grinding: grinding the elm pod sample into fine powder to increase extraction efficiency, and manually grinding with a mortar and pestle;

s1.3: sample particle size screening: the elm pod particles with specific granularity range can be screened out according to the requirement;

s1.4: and (3) storing materials: storing the treated elm pod sample in a sealed container to avoid moisture and light.

3. The method for extracting total flavonoids from elm seeds according to claim 2, wherein S2 is further expressed as:

s2.1: sample selection: selecting dry and fresh elm pod as the raw material for extraction;

s2.2: precooling: the elm pod samples are placed into a refrigerator or a freezing chamber for precooling treatment, which is helpful for sample hardening and reduces heat generation in the crushing process;

s2.3: grinding equipment selection: selecting a suitable grinding apparatus such as a mortar and pestle, a ball mill, or a high speed mixer, etc.;

s2.4: sample pretreatment: depending on the hardness and shape of the elm pod, the sample may be first subjected to rough crushing treatment, such as cutting, crushing or grinding;

s2.5: grinding a small sample: about 10 g to 20 g of the elm pod sample was placed in a mortar and manually ground using a pestle;

s2.6: grinding: grinding the sample with a pestle or a suitable device, and applying force uniformly during manual grinding to perform continuous and repeated up-and-down grinding;

s2.7: and (3) quality control: after grinding, the quality control can be carried out on the crushed sample, and the fineness and uniformity of the sample can be checked.

4. The method of extracting total flavonoids from elm seeds as in claim 3, wherein S3 is further expressed as:

s3.1: selecting a suitable solvent: selecting a suitable organic solvent, typically ethanol or an ethanol water mixture;

s3.2: mixing the sample with a solvent: mixing the crushed elm pod sample with a selected solvent according to a certain proportion;

s3.3: leaching: placing the mixture into a mortar, stirring with a stirrer or pestle to ensure that the sample is fully contacted with the solvent, and sealing the container for leaching;

s3.4: and (3) filtering: filtering the extract with filter paper or filter membrane after leaching, and removing solid particles to obtain elm pod extract;

s3.5: concentrating: if necessary, vacuum concentrating equipment can be used to concentrate the solvent in the extractive solution to obtain concentrated extract;

s3.6: and (3) evaporation: placing the concentrated extract into an evaporation dish, and removing residual solvent by evaporation under safe conditions to obtain flavonoid extract;

s3.7: and (3) quality control: the quality control of the extract can be performed by chromatographic analysis to ensure the purity and purity of the extract.

5. The method of extracting total flavonoids from elm seeds as in claim 4, wherein S4 is further expressed as:

s4.1: preparing elm pod samples: selecting fresh and dried elm pod samples, and crushing the elm pod samples into fine powder according to the requirements;

s4.2: the appropriate solvent is chosen: selecting a proper organic solvent according to the characteristics of flavonoid compounds in elm pod;

s4.3: mixing the sample with a solvent: mixing the crushed elm pod sample with the selected solvent in a certain proportion;

s4.4: leaching time: placing the mixture in a suitable container to avoid light as much as possible, then sealing the container, and leaching for 12-48 hours;

s4.5: and (3) temperature control: the temperature during the leaching process should be controlled within a suitable range, typically between 20 ℃ and 25 ℃;

s4.6: mixing and stirring: during leaching, the mixture was gently stirred periodically to ensure adequate contact of the sample with the solvent;

s4.7: and (3) filtering: filtering the extract with filter paper or filter membrane after leaching, and removing solid particles to obtain elm pod extract;

s4.8: concentrating: if necessary, the solvent in the extract may be concentrated using a vacuum concentration apparatus to obtain a concentrated extract.

6. The method of extracting total flavonoids from elm seeds according to claim 5, wherein S5 is further expressed as:

s5.1: selecting an appropriate filtering device: it is important to select a suitable filtration device;

s5.2: preparing a filtering device: installing the selected filter media into a glass funnel, vacuum funnel or other suitable filter apparatus;

s5.3: filtering the extract: slowly pouring the extract into the filter, pre-moistened filter paper or pre-treated filter may be used for better results;

s5.4: avoid blocking: some solid particles or impurities may be present in the elm pod extract, which may cause clogging of filter paper or filter membrane;

s5.5: collecting the filtered extract: after passing through the filter, the extract is collected in a container as a pure liquid fraction;

s5.6: and (3) re-processing: if a higher purity extract is desired, the filtration step may be repeated or the extract may be subjected to further steps;

s5.7: cleaning the filtering equipment: after the filtration is completed, the used filtration equipment is washed and sterilized.

7. The method of extracting total flavonoids from elm seeds as in claim 6, wherein S6 is further expressed as:

s6.1: preparing a concentration container: selecting an evaporation pan or a burner with proper size;

s6.2: adding an extracting solution: pouring the elm pod total flavone extracting solution into an evaporation container;

s6.3: and (5) heating and concentrating: according to the safety operation rules, the extracting solution is gently heated on a warm water bath or a heating plate, and the temperature is lower than the boiling point of the flavone compound;

s6.4: and (3) periodically stirring: periodically stirring the liquid in the concentrating container;

s6.5: concentrating to the required concentration: heating and stirring are continued until the volume of the extract is reduced to the desired concentration;

s6.6: and (3) cooling: stopping heating once the desired concentration is reached;

s6.7: protection from light or oxygen: if the flavone compounds are sensitive to light or oxygen, a light shielding material may be used on the concentrating container;

s6.8: collecting the concentrate: carefully collecting the concentrated substance into a dry container;

s6.9: measuring the concentration: using a suitable analytical method;

s6.10: and (3) storing: the concentrate is stored under appropriate conditions.

8. The method of extracting total flavonoids from elm seeds as in claim 7, wherein S7 is further expressed as:

s7.1: sample preparation: if a elm pod sample is used, the sample is first ensured to be dry;

s7.2: extracting: mixing the elm pod sample with an organic solvent to soak the sample in the solvent;

s7.3: soaking: allowing the sample to soak in the solvent for a period of time, typically several hours or overnight;

s7.4: and (3) filtering: filtering the extractive solution with screen or filter paper to remove solid particles in elm pod;

s7.5: and (3) evaporation: transferring the extractive solution into evaporating dish or beaker, and slightly heating on warm water bath or heating plate to evaporate organic solvent at a temperature below boiling point of flavone compound, typically 40-60deg.C;

s7.6: and (3) periodically stirring: the liquid in the evaporation pan or beaker is stirred periodically to ensure a uniform evaporation process.

9. The method of extracting total flavonoids from elm seeds according to claim 8, wherein S8 is further expressed as:

s8.1: calibration standard: preparing a series of total flavone standard solutions with different concentrations, and measuring absorbance of elm pods by using an analytical instrument;

s8.2: concentrating to the required concentration: continuing heating and;

s8.3: sample measurement: measuring the content of total flavonoids in the extract by using an analytical instrument;

s8.4: correction and calculation: using a standard curve and analysis results;

s8.5: storing and reporting: the quality-controlled extract is stored under appropriate conditions, typically refrigerated or frozen.

10. The method of extracting total flavonoids from elm seeds as claimed in claim 9, wherein S9 is further expressed as:

s9.1: sample preparation: dissolving the dried extract in a suitable solvent for subsequent analysis;

s9.2: high Performance Liquid Chromatography (HPLC) analysis: analysis of total flavonoids using High Performance Liquid Chromatography (HPLC);

s9.3: standard calibration: using flavonoid standard substances with known concentrations to establish a standard curve;

s9.4: data analysis: analyzing the HPLC result;

s9.5: and (3) quality control: quality control was performed to ensure accuracy and repeatability of the experiment.