Method and system for detecting bioflavonoids in plants

Abstract

The invention discloses a method and a system for detecting bioflavonoids in plants, and relates to the technical field of plant extraction, a plant sample is ground, dissolved and extracted, a sample solution required by detection is obtained, other pigment substances in the sample solution are removed by adopting an acid-base treatment method, and the bioflavonoids in the plant are obtained. The flavonoid substances in the sample solution are purified and enriched through a column chromatography technology; and adjusting the pH value of the sample solution to a pH range capable of maintaining the stability of the flavonoid substances according to the stability requirement of the flavonoid substances. According to the method and system for detecting the bioflavonoids in the plants, through pretreatment, purification and enrichment and pH adjustment, the detection precision and reliability are improved, the purity of a sample solution is ensured, the concentration of flavonoid substances is further improved through a column chromatography technology, the influence of interfering substances is reduced, the sample solution is maintained within the stable pH range of the flavonoid substances, and the detection accuracy is improved. Degradation or conversion is avoided, and the detection accuracy is further improved.

Description

Technical Field

[0001] The invention relates to the technical field of plant extraction, and in particular to a method and system for detecting bioflavonoids in plants. Background Art

[0002] Plant flavonoids have broad application prospects in the medical field. They have multiple pharmacological effects such as anti-inflammatory, antioxidant, and anti-tumor, and are of great significance for the prevention and treatment of various diseases. By detecting the flavonoid content in plant samples, plant resources with medicinal value can be screened out to provide a scientific basis for drug research and development. In addition, flavonoid detection can also be used to evaluate the quality of Chinese medicinal materials and ensure the efficacy and safety of Chinese medicine preparations. At present, the detection system and method of plant flavonoids are already quite mature, including high performance liquid chromatography (HPLC), biochemical method, ultraviolet spectroscopy and other technical means. These technologies are efficient and precise and can accurately determine the flavonoid content in plant samples.

[0003] In the prior art, ultraviolet spectroscopy is simple to operate and takes a short time, but it is easily interfered by other pigment substances, which makes the measured values ​​often too high, resulting in inaccurate test results. Therefore, how to weaken the interference of other pigment substances in plant extracts to improve the detection accuracy of bioflavonoids in plants is a problem we need to solve. To this end, a method and system for detecting bioflavonoids in plants are proposed. Summary of the invention

[0004] To achieve the above objectives, the present invention is implemented by the following technical solutions: In the first aspect, a method for detecting bioflavonoids in plants comprises the following steps: Step 1: Grind, dissolve and extract the plant sample to obtain the sample solution required for detection, use acid-base treatment method to remove other pigment substances in the sample solution, and purify and enrich the flavonoids in the sample solution by column chromatography technology; Step 2: According to the stability requirements of flavonoids, the pH value of the sample solution is adjusted to a pH range that can maintain the stability of flavonoids; Step 3: Scan the sample solution using an ultraviolet spectrometer to obtain its ultraviolet-visible absorption spectrum, and determine the presence of flavonoids based on their characteristic absorption peaks and calculate their absorbance; Step 4: By comparing the absorbance difference between the sample solution containing flavonoids and the sample solution without flavonoids, the influence of the interfering substances on the test results is calculated and corrected; Step 5: Process and analyze the collected data, including peak identification, quantitative calculation and standard curve drawing, and perform quality control on the detection process by comparing and analyzing the performance and stability of the detection process with the standard data.

[0005] In a second aspect, a detection system for bioflavonoids in plants is used to implement a detection method for bioflavonoids in plants, comprising a detection management center, wherein the detection management center is communicatively connected to a sample pretreatment module, a purification and enrichment module, a pH adjustment module, an ultraviolet spectrum detection module, an interference correction module, and a data processing module, wherein electrical signals are connected between the modules; The sample pretreatment module is used to grind, dissolve and extract the plant sample to obtain the sample solution required for detection, and adopt an acid-base treatment method to remove other pigment substances in the sample solution. Through appropriate pretreatment, the interference of other pigments on the ultraviolet spectroscopy is effectively reduced, and the detection accuracy of flavonoids is improved; The purification and enrichment module purifies and enriches the flavonoids in the sample solution through column chromatography technology, thereby increasing the concentration of the flavonoids, further reducing the impact of interfering substances, and improving the sensitivity of detection; The pH adjustment module adjusts the pH value of the sample solution to maintain the sample solution within a stable pH range for flavonoids, thereby preventing the flavonoids from being degraded or transformed during the detection process and improving the accuracy of the detection; The ultraviolet spectrum detection module uses an ultraviolet spectrometer to scan the sample solution, uses ultraviolet-visible spectrophotometry to detect the characteristic absorption peaks of flavonoids, and calculates the content of flavonoids in the original sample based on the relationship between absorbance and flavonoid concentration; The interference correction module constructs an interference correction model, compares the absorbance difference between sample solutions containing flavonoids and those without flavonoids, calculates the influence of interfering substances on the detection results, and then corrects the interference in the detection process, reduces the influence of other pigment substances on the detection of flavonoid compounds, and improves the reliability of the detection results; The data processing module is used to process and analyze the collected data, including peak value identification, quantitative calculation and standard curve drawing, to improve the accuracy and reliability of the data and provide a basis for subsequent data analysis and comparison.

[0006] Preferably, in the sample preprocessing module, the process of obtaining the sample solution includes: The fresh plant samples are cleaned and chopped, and then dried to obtain dried plant samples, wherein the fresh plant samples are divided into fruits and vegetables and medicinal plants, the fruits and vegetables include citrus, grapes, apples, onions and blueberries, and the medicinal plants include ginkgo leaves, sophora japonica, astragalus, kudzu root and epimedium, etc.; Grinding the plant dry sample to destroy its cell structure, and adding the ground sample into an organic solvent to dissolve and extract the flavonoids according to the solubility of the flavonoids, wherein the organic solvent is methanol or ethanol; The dissolved and extracted sample solution is filtered using a filter membrane to remove residues and insoluble substances, and the pH value of the sample solution is adjusted using an acid-base reagent to remove other pigment substances in the sample solution; Through acid-base treatment, the pigment substances in the sample solution are precipitated, dissolved or changed in color, which is convenient for subsequent removal. The treated sample solution is fixed to a certain volume to obtain a sample solution that meets the detection requirements.

[0007] Preferably, in the purification and enrichment module, the process of increasing the concentration of flavonoids comprises: Prepare the polyamide column chromatography material required for column chromatography, select gel material as filler material, and fill the selected filler evenly into the column. During the filling process, pay attention to avoid the generation of gaps and bubbles to ensure that the sample solution can pass through the filler evenly. Among them, polyamide has a good separation effect on various phenolic hydroxyl flavonoid compounds (including glycosides and aglycones), and its chromatography capacity is relatively large; The treated sample solution is dripped onto the top of the polyamide column, so that the sample solution slowly penetrates into the column chromatography material to form a wet loading method. For the polyamide column, water and ethanol are selected for gradient elution. During the elution process, flavonoids are gradually separated according to the difference in distribution coefficients in the stationary phase and the mobile phase. During column chromatography, there is a certain dead volume before the target molecule elutes. The size of the dead volume depends on the size of the column and the separated compound. The starting solvent level and the mark corresponding to the length of the silica gel below are marked on the polyamide column. Once the solvent level drops below the mark, fraction collection can begin. The fraction containing flavonoids is collected separately. Based on the test results, the fractions containing the target flavonoids were determined and combined for further purification and analysis. The purification and enrichment of the target molecules were achieved by adjusting the column chromatography parameters (eluent concentration, flow rate, etc.).

[0008] Preferably, in the pH adjustment module, the process of adjusting the pH value of the sample solution includes: Obtain the sample solution to be tested, ensure that it is pure and suitable for pH adjustment, use a pH meter to measure the initial pH value of the sample solution, and determine the target pH range based on the properties of flavonoids and the test requirements. Flavonoids show better stability within a specific pH range, where the target pH value of flavonoids is 5-6; According to the difference between the initial pH value and the target pH value range, use a burette to add acid or base to the sample solution to adjust the pH value of the sample solution. If the sample solution is acidic, add sodium carbonate as an alkaline regulator. If the sample solution is alkaline, add hydrochloric acid as an acidic regulator. During the adjustment process, the pH meter is continuously used to monitor the pH change of the solution until a predetermined target pH value range is reached. When the target pH value range is reached, a buffer is added to stabilize the pH of the solution to prevent pH changes in subsequent operations, wherein the buffer is a phosphate buffer or an acetate buffer; After pH adjustment, verify the stability of the sample solution by measuring the pH value again after a period of time (24 hours) to check if there is any change, and record the pH value and the adjuster used for each adjustment to facilitate subsequent data analysis and comparison.

[0009] Preferably, in the ultraviolet spectrum detection module, the calculation process of the flavonoid content in the original sample includes: Ensure that the sample solution has been pH adjusted and purified and enriched, and is in the required state for UV spectroscopy detection. According to the characteristic absorption spectrum of flavonoids, select the wavelength range with the absorption peak between 200-400nm for scanning; Use samples of known concentration to calibrate the UV spectrometer, dilute the sample solution to a concentration that is suitable for the detection range of the UV spectrometer, put the sample solution into a quartz cuvette, and put the cuvette into the sample slot of the UV spectrometer; A blank solvent is used as a background to perform a background scan to eliminate interference from the solvent and other non-sample substances, and a UV spectrum obtained by the scan is recorded. Based on the known properties of flavonoids, characteristic absorption peaks are identified in the UV spectrum, wherein flavonoids have one or more characteristic absorption peaks in the range of 240-370 nm; At the characteristic absorption peak, measure the absorbance value of the sample solution to ensure the consistency of the measurement conditions (wavelength, slit width, light source intensity, etc.), and use flavonoid standards of known concentration to establish a linear relationship between absorbance and concentration (standard curve); According to the absorbance value of the sample solution, the corresponding flavonoid concentration is found on the standard curve, and the flavonoid content in the original sample is calculated based on the dilution multiple of the sample solution.

[0010] Preferably, the calculation formula for the flavonoid content in the original sample is: ; in, is the concentration of flavonoids in the original sample, is the absorbance value of the sample solution at the characteristic absorption peak, is the molar absorption coefficient, which indicates the absorption capacity of a substance to light at a specific wavelength. is the optical path length of the cuvette, usually 1 cm, is the absorbance value corresponding to a specific concentration on the standard curve.

[0011] Preferably, in the interference correction module, the calculation process of the influence of the interfering substance on the detection result includes: Obtain sample solutions containing flavonoids and sample solutions without flavonoids, ensure that both have undergone the same pretreatment and purification and enrichment steps, and scan the two sample solutions using an ultraviolet spectrometer to record the absorbance values ​​within a wavelength range of 200-400 nm; Collect the absorbance of the sample solution containing flavonoids and the sample solution without flavonoids, compare the absorbance difference between the sample solution containing flavonoids and the sample solution without flavonoids, and identify the wavelength point where significant absorbance appears in the non-flavonoid sample, which corresponds to the absorption peak of the interfering substance; The interference correction model was constructed by using multiple linear regression, with the absorbance value of non-flavonoid samples as the independent variable and the additional absorbance value caused by interfering substances in flavonoid samples as the dependent variable. For the flavonoid sample to be tested, the absorbance value is measured by ultraviolet spectrometer, and the additional absorbance value caused by the interfering substance is calculated by using the constructed interference correction model, and the corrected absorbance value is obtained by combining the original absorbance value and the additional absorbance value; The concentration of flavonoids in the sample to be tested was calculated using the corrected absorbance value in combination with the established standard curve (linear relationship between absorbance and flavonoid concentration).

[0012] Preferably, the calculation formula of the corrected absorbance value is: ; in, is the corrected absorbance value, is the absorbance value of the sample solution at the characteristic absorption peak, It is The interference coefficient of each interfering substance indicates the influence of the interfering substance on the absorbance of flavonoids. It is The absorbance value of the interfering substance in the sample, that is, the absorbance value corresponding to the wavelength point where significant absorbance appears in the non-flavonoid sample, is the total number of interfering substances, indicating how many interfering substances are present in the sample. is the absorbance value of the standard sample, used for normalization, from a standard of known concentration.

[0013] Preferably, the calculation formula for the concentration of flavonoids in the sample to be tested is: ; in, is the concentration of flavonoids in the sample to be tested, is the corrected absorbance value, is the molar absorption coefficient, which indicates the absorption capacity of a substance to light at a specific wavelength. It is a constant. is the optical path length of the cuvette, is the absorbance value of the standard sample, which is used for normalization and to establish a standard curve. is the tuning index used to adjust the characteristics of the formula to fit the data.

[0014] The present invention provides a method and system for detecting bioflavonoids in plants, which have the following beneficial effects: 1. The detection method and system of bioflavonoids in the plant improve the accuracy and reliability of detection through pretreatment, purification and enrichment, and pH adjustment, ensure the purity of the sample solution, and further increase the concentration of flavonoids through column chromatography technology, reduce the influence of interfering substances, maintain the sample solution within the stable pH range of flavonoids, avoid degradation or conversion, and further improve the accuracy of detection.

[0015] 2. The detection method and system of bioflavonoids in the plant constructs an interference correction model, compares the absorbance difference between sample solutions containing flavonoids and those not containing flavonoids, calculates the influence of interfering substances on the detection results, and performs correction, thereby reducing the influence of other pigment substances on the detection of flavonoid compounds and improving the reliability of the detection results. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 is a flow chart of the method of the present invention; Figure 2 It is a system module diagram of the present invention; Figure 3 is a flow chart for calculating the content of flavonoids in the original sample of the present invention; Figure 4 The figure is a flow chart of calculation of the influence of interfering substances on the detection results of the present invention. DETAILED DESCRIPTION

[0017] The present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. The embodiments of the present invention are provided for the purpose of illustration and description, and are not intended to be exhaustive or to limit the present invention to the disclosed forms. Many modifications and variations will be apparent to those of ordinary skill in the art. The embodiments are selected and described in order to better illustrate the principles and practical applications of the present invention, and to enable those of ordinary skill in the art to understand the present invention and thereby design various embodiments with various modifications suitable for specific uses.

[0018] The first embodiment, as Figure 1 As shown, the present invention provides a technical solution: a method for detecting bioflavonoids in plants, comprising the following steps: Step 1: Grind, dissolve and extract the plant sample to obtain the sample solution required for detection, use acid-base treatment method to remove other pigment substances in the sample solution, and purify and enrich the flavonoids in the sample solution by column chromatography technology; Step 2: According to the stability requirements of flavonoids, the pH value of the sample solution is adjusted to a pH range that can maintain the stability of flavonoids; Step 3: Scan the sample solution using an ultraviolet spectrometer to obtain its ultraviolet-visible absorption spectrum, and determine the presence of flavonoids based on their characteristic absorption peaks and calculate their absorbance; Step 4: By comparing the absorbance difference between the sample solution containing flavonoids and the sample solution without flavonoids, the influence of the interfering substances on the test results is calculated and corrected; Step 5: Process and analyze the collected data, including peak identification, quantitative calculation and standard curve drawing, and perform quality control on the detection process by comparing and analyzing the performance and stability of the detection process with the standard data.

[0019] The second embodiment is based on the first embodiment. Figure 2 , Figure 3 As shown, the present invention also provides a technical solution: a detection system for bioflavonoids in plants, used to implement a detection method for bioflavonoids in plants, including a detection management center, the detection management center is communicatively connected to a sample pretreatment module, a purification and enrichment module, a pH adjustment module, an ultraviolet spectrum detection module, an interference correction module and a data processing module, wherein the modules are connected by electrical signals; The sample pretreatment module is used to grind, dissolve and extract plant samples to obtain the sample solution required for detection, and use acid-base treatment methods to remove other pigment substances in the sample solution. Through appropriate pretreatment, the interference of other pigments on ultraviolet spectroscopy can be effectively reduced, and the detection accuracy of flavonoids can be improved. The fresh plant samples are washed and chopped, and then dried to obtain dry plant samples. Among them, the fresh plant samples are divided into fruits and vegetables and medicinal plants. Fruits and vegetables include citrus, grapes, apples, onions and blueberries, and medicinal plants include ginkgo leaves, Sophora japonica, Astragalus, Pueraria and Epimedium, etc. The dried sample of the plant is ground to destroy its cell structure, and according to the solubility of the flavonoids, the ground sample is added to an organic solvent to dissolve and extract it, wherein the organic solvent is methanol or ethanol, and the sample solution after dissolution and extraction is filtered using a filter membrane to remove residues and insoluble substances, and the pH value of the sample solution is adjusted using an acid-base reagent to remove other pigment substances in the sample solution. The pigment substances in the sample solution are precipitated, dissolved or changed in color through acid-base treatment, so as to facilitate subsequent removal, and the treated sample solution is fixed to a volume to obtain a sample solution that meets the detection requirements; The purification and enrichment module purifies and enriches the flavonoids in the sample solution through column chromatography technology, increases the concentration of flavonoids, further reduces the influence of interfering substances, and improves the sensitivity of detection. The polyamide column chromatography material required for column chromatography is configured, and the gel material is selected as the filler material. The selected filler is evenly filled into the column. During the filling process, attention is paid to avoid the generation of gaps and bubbles to ensure that the sample solution can pass through the filler evenly. Among them, polyamide has a good separation effect on various phenolic hydroxyl flavonoid compounds (including glycosides and aglycones), and its chromatography capacity is relatively large. The treated sample solution is dripped onto the top of the polyamide column, so that the sample solution slowly penetrates into the column chromatography material to form a wet loading method. For the polyamide column, water and ethanol are used for gradient elution. During the elution process, flavonoids are gradually separated according to the difference in distribution coefficients in the stationary phase and the mobile phase. During the column chromatography process, there is a certain dead volume before the target molecule flows out. The size of the dead volume depends on the size of the column and the separated compound. The starting solvent level and the mark corresponding to the length of the silica gel below are marked on the polyamide column. Once the solvent level drops below the mark, the fractions can be collected. The fractions containing flavonoids are collected separately. According to the test results, the fractions containing the target flavonoids are determined and combined for further purification and analysis. The purification and enrichment of the target molecule can be achieved by adjusting the column chromatography parameters (eluent concentration, flow rate, etc.); The pH adjustment module adjusts the pH value of the sample solution to maintain the sample solution within the stable pH range of flavonoids, avoids degradation or conversion of flavonoids during the detection process, improves the accuracy of the detection, obtains the sample solution to be detected, ensures that it is pure and suitable for pH adjustment, uses a pH meter to measure the initial pH value of the sample solution, and determines the target pH range based on the properties of flavonoids and detection requirements. Flavonoids show better stability within a specific pH range, among which the target pH value of flavonoids is 5-6. Based on the difference between the initial pH value and the target pH range, use a burette to add acid or alkali to the sample solution to adjust the sample The pH value of the solution. If the sample solution is acidic, sodium carbonate is added as an alkaline regulator. If the sample solution is alkaline, hydrochloric acid is added as an acidic regulator. During the adjustment process, a pH meter is continuously used to monitor the pH change of the solution until the predetermined target pH range is reached. When the target pH range is reached, a buffer is added to stabilize the pH of the solution to prevent pH changes in subsequent operations. The buffer is a phosphate buffer or an acetate buffer. After the pH is adjusted, the stability of the sample solution is verified. The pH value is measured again after a period of time (24 hours) to check whether there is any change. The pH value and the regulator used for each adjustment are recorded to facilitate subsequent data analysis and comparison; The UV spectrum detection module uses a UV spectrometer to scan the sample solution, uses UV-visible spectrophotometry to detect the characteristic absorption peaks of flavonoids, and calculates the content of flavonoids in the original sample based on the relationship between absorbance and flavonoid concentration. Ensure that the sample solution has been pH-adjusted and purified and enriched, and is in the required state for UV spectrum detection. According to the characteristic absorption spectrum of flavonoids, select a wavelength range with an absorption peak between 200-400nm for scanning, use a sample of known concentration to calibrate the UV spectrometer, dilute the sample solution to a concentration that is suitable for the detection range of the UV spectrometer, and place the sample solution in a quartz cuvette. The cuvette is placed in the sample slot of the UV spectrometer, and a blank solvent is used as a Perform background scanning for background to eliminate interference from solvents and other non-sample substances, record the ultraviolet spectrum obtained by scanning, and identify characteristic absorption peaks in the ultraviolet spectrum according to the known characteristics of flavonoids, wherein flavonoids have one or more characteristic absorption peaks in the range of 240-370nm. At the characteristic absorption peak, measure the absorbance value of the sample solution to ensure the consistency of measurement conditions (wavelength, slit width, light source intensity, etc.), and use flavonoid standards of known concentration to establish a linear relationship between absorbance and concentration (standard curve). According to the absorbance value of the sample solution, find the corresponding flavonoid concentration on the standard curve, and calculate the flavonoid content in the original sample based on the dilution multiple of the sample solution; Furthermore, the calculation formula for the flavonoid content in the original sample is: ; in, is the concentration of flavonoids in the original sample, is the absorbance value of the sample solution at the characteristic absorption peak, is the molar absorption coefficient, which indicates the absorption capacity of a substance to light at a specific wavelength. is the optical path length of the cuvette, usually 1 cm, is the absorbance value corresponding to a specific concentration on the standard curve, serving as a reference point. and The value range of depends on the measurement range of the UV spectrometer used in the experiment. Usually the absorbance value is between 0 and 2. Indicates the effect of absorbance deviation on concentration calculation. near When , the root part is close to 1, indicating that the absorbance measurement value is close to the predicted value of the standard curve; when and When the difference is large, the radical part increases, indicating that there is a large deviation. The influence of absorbance deviation on concentration calculation was further considered. equal When , the exponential function value is 1, indicating no deviation; when and The larger the difference, the greater the exponential function value, indicating a greater deviation; The interference correction module builds an interference correction model, compares the absorbance difference between sample solutions containing flavonoids and those without flavonoids, calculates the impact of interfering substances on the test results, and then corrects the interference in the test process, reduces the impact of other pigment substances on the detection of flavonoid compounds, and improves the reliability of the test results; The data processing module is used to process and analyze the collected data, including peak recognition, quantitative calculation and standard curve drawing, to improve the accuracy and reliability of the data and provide a basis for subsequent data analysis and comparison.

[0020] The third embodiment, based on the second embodiment, please refer to Figure 4 As shown, in the interference correction module, the calculation process of the influence of interfering substances on the test results includes: Obtain sample solutions containing flavonoids and those without flavonoids, ensure that both have undergone the same pretreatment and purification enrichment steps, and use an ultraviolet spectrometer to scan the two sample solutions, record the absorbance values ​​in the wavelength range of 200-400nm, collect the absorbance of the sample solutions containing flavonoids and the sample solutions without flavonoids, compare the absorbance differences between the sample solutions containing flavonoids and those without flavonoids, identify the wavelength points where significant absorbance appears in the non-flavonoid samples, corresponding to the absorption peaks of the interfering substances, and use multiple linear regression to Taking the absorbance value of non-flavonoid samples as the independent variable and the additional absorbance value caused by interfering substances in flavonoid samples as the dependent variable, an interference correction model is constructed. For the flavonoid samples to be tested, the absorbance value is measured by using an ultraviolet spectrometer. The additional absorbance value caused by interfering substances is calculated using the constructed interference correction model, and the corrected absorbance value is obtained by combining the original absorbance value and the additional absorbance value. The corrected absorbance value is used in combination with the established standard curve (linear relationship between absorbance and flavonoid concentration) to calculate the concentration of flavonoids in the sample to be tested; Furthermore, the calculation formula of the corrected absorbance value is: ; in, is the corrected absorbance value, is the absorbance value of the sample solution at the characteristic absorption peak, It is The interference coefficient of each interfering substance indicates the influence of the interfering substance on the absorbance of flavonoids. It is The absorbance value of the interfering substance in the sample, that is, the absorbance value corresponding to the wavelength point where significant absorbance appears in the non-flavonoid sample, is the total number of interfering substances, indicating how many interfering substances are present in the sample. is the absorbance value of the standard sample, used for normalization, from a standard of known concentration, Indicates the effect of absorbance deviation on the calibration result. When the deviation is larger, the value of the root part increases, resulting in a decrease in the absorbance value after correction. The influence of interfering substances on absorbance was further considered. When the absorbance value of interfering substances increased, the exponential function value decreased, resulting in a decrease in the corrected absorbance value. Furthermore, the calculation formula for the concentration of flavonoids in the sample to be tested is: ; in, is the concentration of flavonoids in the sample to be tested, is the corrected absorbance value, is the molar absorption coefficient, which indicates the absorption capacity of a substance to light at a specific wavelength. It is a constant. is the optical path length of the cuvette, is the absorbance value of the standard sample, which is used for normalization and to establish a standard curve. It is an adjustment index used to adjust the characteristics of the formula fitting data, comprehensively consider the relationship between the absorbance value and the concentration of flavonoids, and the influence of the corrected absorbance value on the concentration calculation, so as to obtain a more accurate flavonoid content.

[0021] Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field and related fields without creative work should fall within the scope of protection of the present invention. The structures, devices and operating methods not specifically described and explained in the present invention are implemented according to the conventional means in the field unless otherwise specified and limited.

Claims

1. A method for detecting bioflavonoids in plants, characterized in that: The following steps are involved: Step 1: Grind, dissolve and extract the plant sample to obtain the sample solution required for detection, use acid-base treatment method to remove other pigment substances in the sample solution, and purify and enrich the flavonoids in the sample solution by column chromatography technology; Step 2: According to the stability requirements of flavonoids, the pH value of the sample solution is adjusted to a pH range that can maintain the stability of flavonoids; Step 3: Scan the sample solution using an ultraviolet spectrometer to obtain its ultraviolet-visible absorption spectrum, and determine the presence of flavonoids based on their characteristic absorption peaks and calculate their absorbance; Step 4: By comparing the absorbance difference between the sample solution containing flavonoids and the sample solution without flavonoids, the influence of the interfering substances on the test results is calculated and corrected; Step 5: Process and analyze the collected data, and perform quality control on the detection process by comparing and analyzing the performance and stability of the detection process with the standard data.

2. A system for detecting bioflavonoids in plants, used to implement the method for detecting bioflavonoids in plants as claimed in claim 1, comprising a detection management center, characterized in that: The detection management center is communicatively connected with a sample pretreatment module, a purification and enrichment module, a pH adjustment module, an ultraviolet spectrum detection module, an interference correction module and a data processing module, wherein electrical signals are connected between the modules; The sample pretreatment module is used to grind, dissolve and extract the plant sample to obtain the sample solution required for detection, and use an acid-base treatment method to remove other pigment substances in the sample solution; The purification and enrichment module purifies and enriches the flavonoids in the sample solution by column chromatography technology to increase the concentration of the flavonoids; The pH adjustment module adjusts the pH value of the sample solution so that the sample solution is maintained within a pH range where flavonoids are stable; The ultraviolet spectrum detection module uses an ultraviolet spectrometer to scan the sample solution, uses ultraviolet-visible spectrophotometry to detect the characteristic absorption peaks of flavonoids, and calculates the content of flavonoids in the original sample based on the relationship between absorbance and flavonoid concentration; The interference correction module constructs an interference correction model, compares the absorbance difference between sample solutions containing flavonoids and sample solutions without flavonoids, calculates the effect of interfering substances on the test results, and then corrects the interference in the test process; The data processing module is used to process and analyze the collected data, including peak value identification, quantitative calculation and standard curve drawing.

3. The detection system of bioflavonoids in plants according to claim 2, characterized in that: In the sample pretreatment module, the process of obtaining the sample solution includes: The fresh plant samples are washed and chopped, and then dried to obtain dried plant samples, wherein the fresh plant samples are divided into fruits and vegetables and medicinal plants, the fruits and vegetables include citrus, grapes, apples, onions and blueberries, and the medicinal plants include ginkgo leaves, sophora japonica seeds, astragalus, kudzu root and epimedium; Grinding the plant dry sample to destroy its cell structure, and adding the ground sample into an organic solvent to dissolve and extract the flavonoids according to the solubility of the flavonoids, wherein the organic solvent is methanol or ethanol; The dissolved and extracted sample solution is filtered using a filter membrane to remove residues and insoluble substances, and the pH value of the sample solution is adjusted using an acid-base reagent to remove other pigment substances in the sample solution; Through acid-base treatment, the pigment substances in the sample solution are precipitated, dissolved or the color changes, and the treated sample solution is fixed to a certain volume to obtain a sample solution that meets the detection requirements.

4. The detection system of bioflavonoids in plants according to claim 3, characterized in that: In the purification and enrichment module, the process of increasing the concentration of flavonoids includes: Prepare the polyamide column chromatography material required for column chromatography, select the gel material as the filler material, and evenly fill the selected filler into the column; The treated sample solution is dripped onto the top of the polyamide column, so that the sample solution slowly penetrates into the column chromatography material to form a wet loading method. For the polyamide column, water and ethanol are selected for gradient elution. During the elution process, flavonoids are gradually separated according to the difference in distribution coefficients in the stationary phase and the mobile phase. During column chromatography, there is a certain dead volume before the target molecule elutes. The size of the dead volume depends on the size of the column and the separated compound. The starting solvent level and the mark corresponding to the length of the silica gel below are marked on the polyamide column. Once the solvent level drops below the mark, fraction collection can begin. The fraction containing flavonoids is collected separately. Based on the test results, the fractions containing the target flavonoids were determined and combined for further purification and analysis. The purification and enrichment of the target molecules were achieved by adjusting the column chromatography parameters.

5. The detection system of bioflavonoids in plants according to claim 4, characterized in that: In the pH adjustment module, the pH adjustment process of the sample solution includes: Obtain a sample solution to be tested, use a pH meter to measure the initial pH value of the sample solution, and determine the target pH value range according to the properties of the flavonoids and the test requirements, wherein the target pH value of the flavonoids is 5-6; According to the difference between the initial pH value and the target pH value range, use a burette to add acid or base to the sample solution to adjust the pH value of the sample solution. If the sample solution is acidic, add sodium carbonate as an alkaline regulator. If the sample solution is alkaline, add hydrochloric acid as an acidic regulator. During the adjustment process, the pH meter is continuously used to monitor the pH change of the solution until a predetermined target pH value range is reached, and when the target pH value range is reached, a buffer is added to stabilize the pH of the solution, wherein the buffer is a phosphate buffer or an acetate buffer; After pH adjustment, verify the stability of the sample solution and record the pH value and the adjusting agent used for each adjustment.

6. The detection system of bioflavonoids in plants according to claim 5, characterized in that: In the ultraviolet spectrum detection module, the calculation process of the flavonoid content in the original sample includes: Ensure that the sample solution has been pH adjusted and purified and enriched, and is in the required state for UV spectroscopy detection. According to the characteristic absorption spectrum of flavonoids, select the wavelength range with the absorption peak between 200-400nm for scanning; Use samples of known concentration to calibrate the UV spectrometer, dilute the sample solution to a concentration that is suitable for the detection range of the UV spectrometer, put the sample solution into a quartz cuvette, and put the cuvette into the sample slot of the UV spectrometer; Using a blank solvent as a background to perform background scanning, recording the ultraviolet spectrum obtained by the scanning, and identifying characteristic absorption peaks in the ultraviolet spectrum according to known characteristics of flavonoids, wherein flavonoids have one or more characteristic absorption peaks in the range of 240-370nm; At the characteristic absorption peak, the absorbance value of the sample solution is measured, and a linear relationship between absorbance and concentration is established using a flavonoid standard with a known concentration; According to the absorbance value of the sample solution, the corresponding flavonoid concentration is found on the standard curve, and the flavonoid content in the original sample is calculated based on the dilution multiple of the sample solution.

7. A system for detecting bioflavonoids in plants according to claim 6, characterized in that: The calculation formula for the flavonoid content in the original sample is: ； in, is the concentration of flavonoids in the original sample, is the absorbance value of the sample solution at the characteristic absorption peak, is the molar absorption coefficient, which indicates the absorption capacity of a substance to light at a specific wavelength. is the optical path length of the cuvette, is the absorbance value corresponding to a specific concentration on the standard curve.

8. The detection system of bioflavonoids in plants according to claim 7, characterized in that: In the interference correction module, the calculation process of the influence of interfering substances on the detection results includes: Obtaining sample solutions containing flavonoids and sample solutions without flavonoids, scanning the two sample solutions using an ultraviolet spectrometer, and recording absorbance values ​​within a wavelength range of 200-400 nm; Collect the absorbance of the sample solution containing flavonoids and the sample solution without flavonoids, compare the absorbance difference between the sample solution containing flavonoids and the sample solution without flavonoids, and identify the wavelength point where significant absorbance appears in the non-flavonoid sample, which corresponds to the absorption peak of the interfering substance; The interference correction model was constructed by using multiple linear regression, with the absorbance value of non-flavonoid samples as the independent variable and the additional absorbance value caused by interfering substances in flavonoid samples as the dependent variable. For the flavonoid sample to be tested, the absorbance value is measured by ultraviolet spectrometer, and the additional absorbance value caused by the interfering substance is calculated by using the constructed interference correction model, and the corrected absorbance value is obtained by combining the original absorbance value and the additional absorbance value; The concentration of flavonoids in the sample to be tested was calculated using the corrected absorbance value in combination with the established standard curve.

9. The system for detecting bioflavonoids in plants according to claim 8, characterized in that: The calculation formula of the corrected absorbance value is: ； in, is the corrected absorbance value, is the absorbance value of the sample solution at the characteristic absorption peak, It is The interference coefficient of each interfering substance indicates the influence of the interfering substance on the absorbance of flavonoids. It is The absorbance value of the interfering substance in the sample, is the total number of interfering substances, is the absorbance value of the standard sample, which is used for normalization.

10. A system for detecting bioflavonoids in plants according to claim 9, characterized in that: The calculation formula for the concentration of flavonoids in the sample to be tested is: ； in, is the concentration of flavonoids in the sample to be tested, is the corrected absorbance value, is the molar absorptivity, which is a constant. is the optical path length of the cuvette, is the absorbance value of the standard sample, is the adjustment index.

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