Preparation method and quality detection method of standard sophora fruit decoction

Abstract

The invention discloses a preparation method and a quality detection method of a fructus sophorae standard decoction, and relates to the technical field of quality research of traditional Chinese medicinal materials. The method comprises the following steps: weighing a proper amount of fructus sophorae decoction pieces, putting the fructus sophorae decoction pieces into a marmite, decocting twice, adding 8 times of water for soaking for 30 minutes for the first time, boiling with strong fire, and then decocting with slow fire for 30 minutes; adding 6 times of water for the second time, boiling with strong fire, then decocting with slow fire for 20 minutes, filtering the liquid medicine with 200 meshes, cooling the filtrate to room temperature in a cold water bath, combining the two filtrates, carrying out vacuum concentration at the temperature of 50 DEG C, carrying out vacuum concentration at the absolute vacuum degree of 30-40mbar until the weight ratio of the medicinal material to the fluid extract is about 1: 1, collecting the concentrated solution, and carrying out freeze drying to obtain a sample. According to the method, key quality indexes such as the main content, the characteristic chromatogram and the extract of the fructus sophorae after the decoction is prepared are measured, the transfer rate is investigated, the component transfer condition from the decoction pieces to the decoction is measured through objective quality standards, and the quality degree and effectiveness of the decoction are ensured.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine quality research technology, and in particular relates to the preparation of a standard decoction of Sophora japonica fruit and its quality testing method. Background Technology

[0002] Modern research has demonstrated that Sophora japonica fruit contains a variety of chemical components, including flavonoids, isoflavones (sophoratoside), alkaloids, triterpenoid saponins, amino acids, and phospholipids, with particular emphasis on isoflavones and their glycosides. Sophora japonica fruit possesses a variety of pharmacological activities, including sedation, anti-inflammatory and analgesic effects, anticonvulsant and antispasmodic effects, vasoconstriction relief, inhibition of endothelin activity, maintenance of plaque stability, inhibition of nerve cell malnutrition and ischemic brain injury, and induction of cancer cell apoptosis and antitumor effects. Sophoratoside, in particular, exhibits anti-inflammatory, anticancer, and immunosuppressive effects. The 2020 edition of the Chinese Pharmacopoeia lists sophoratoside as an indicator component for the quality control of Sophora japonica fruit, and the "Quality Standard for Sophora japonica Fruit Formula Granules" YBZ-PFKL-2021062 specifies sophoratoside as a content quality control component. Therefore, this study selected sophoratoside as the quality control indicator component for the standard Sophora japonica fruit decoction.

[0003] The clinical use of traditional Chinese medicine (TCM) is primarily based on decoctions. The material basis of TCM decoctions forms the foundation for disease prevention and treatment under the guidance of TCM theory. Current legal standards only quantitatively control single components, and the dose-effect relationship cannot fully reflect the overall effect of TCM components. At present, given that the vast majority of effective components in TCM are not clearly identified, the establishment of TCM fingerprinting / characteristic maps can greatly improve the technical level and scientific content of TCM quality control.

[0004] Current literature reports on fingerprint chromatograms of Sophora japonica fruit mostly use conventional HPLC methods, and all of them only target the material basis of the raw medicinal material. The index components are mostly fat-soluble components. They are mainly used for qualitative identification of the authenticity, origin and quality differences of Chinese medicinal materials, and cannot fully reflect the material basis characteristics of Chinese medicine decoctions. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing a standard decoction of Sophora japonica fruit and for testing its quality. By measuring key quality indicators such as the main content, characteristic spectrum, and extract of Sophora japonica fruit after decoction preparation, the transfer rate was investigated, the range of the transfer rate of the main content of the medicinal material was judged, and the effectiveness of the decoction was tested, thus solving the problem that it is difficult to evaluate and control the efficacy and quality of existing Sophora japonica fruit decoctions.

[0006] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution:

[0007] This invention relates to the preparation of a standard decoction of Sophora japonica fruit, comprising the following steps:

[0008] Step S1: Weigh out the Sophora japonica fruit slices, place them in a clay pot, and decoct twice;

[0009] Step S2: Add 8 times the amount of water, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes.

[0010] Step S3: Add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 20 minutes;

[0011] Step S4: Filter the solution through a 200-mesh sieve, cool the filtrate to room temperature in a cold water bath, and combine the two filtrates;

[0012] Step S5: Cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate them under vacuum at 50°C;

[0013] Step S6: Concentrate to a fluid extract with a weight ratio of approximately 1:1 between the medicinal material and the fluid extract;

[0014] Step S7: Collect the concentrated solution and freeze-dry it to obtain the sample;

[0015] Step S8: Calculate and measure the indicators of Chinese medicine in the decoction.

[0016] As a preferred technical solution, in step S8, the aqueous extract of Sophora japonica is concentrated under vacuum, and the concentrate is freeze-dried to dryness. The dried extract powder is collected, weighed, and the yield is calculated. The formula for the yield is as follows:

[0017]

[0018] This invention relates to a quality testing method for a standard decoction of Sophora japonica fruit, comprising the following steps:

[0019] Step A, Preparation of the reference solution: Take an appropriate amount of succinic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 0.1 mg per 1 ml.

[0020] Step B: Take an appropriate amount of this product, grind it into a fine powder, take about 0.2g, weigh it accurately, place it in a stoppered conical flask, accurately add 50ml of 50% methanol, weigh it, sonicate for 20 minutes, cool it, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, and take the filtrate to obtain the product.

[0021] Step C, Preparation of reference solution: Take 0.5g of Sophora japonica pod reference material, place it in a stoppered conical flask, add 50ml of 50% methanol, seal tightly, sonicate for 20 minutes, cool, shake well, filter, and take the filtrate as the reference solution of the reference material; separately take an appropriate amount of rutin reference standard, accurately weigh it, add methanol to prepare a solution containing 0.2mg per 1ml, as the reference solution of the reference standard;

[0022] Step D, Setup: Inject 10 μl each of the obtained reference solution, test solution, and standard solution into the liquid chromatograph for detection;

[0023] The conditions used for chromatographic detection in the liquid chromatograph include:

[0024] Chromatographic column: The column is packed with octadecylsilane-bonded silica gel, with a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm;

[0025] Column temperature: 30℃;

[0026] Mobile phase: Acetonitrile as mobile phase A, and 0.1% phosphoric acid as mobile phase B;

[0027] Mobile phase flow rate: 1.2 mL per minute;

[0028] Detection wavelength: 282nm;

[0029] The theoretical plate number, calculated based on the rutin peak, should be no less than 3000;

[0030] The mobile phase is eluted using a gradient program as follows:

[0031] From 0 to 12 min, the mobile phase A increased from 5% to 20%, while the mobile phase B decreased from 95% to 80%.

[0032] From 12 to 25 minutes, the concentration of mobile phase A increased from 20% to 22%, while the concentration of mobile phase B decreased from 80% to 78%.

[0033] Over 25–40 minutes, the mobile phase A concentration increased from 22% to 50%, while the mobile phase B concentration decreased from 78% to 50%.

[0034] As a preferred technical solution, the chromatogram of the test sample includes nine characteristic peaks, which should correspond to the relative retention times of the nine characteristic peaks in the chromatogram of the reference medicinal material; the peak corresponding to the rutin reference material is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 is 0.353, peak 2 is 0.679, peak 3 is 0.844, peak 4 is 0.945, peak 6 is 1.165, peak 7 is 1.220, peak 8 is 1.360, and peak 9 is 1.405.

[0035] The present invention has the following beneficial effects:

[0036] This invention measures key quality indicators such as the main content, characteristic spectrum, and extract of Sophora japonica fruit after decoction preparation, investigates the transfer rate, judges the range of transfer rate of the main content of medicinal materials, and detects the effectiveness of the decoction, thereby improving the technical level and scientific content of traditional Chinese medicine quality control.

[0037] Of course, any product implementing this invention does not necessarily need to achieve all of the advantages described above at the same time. Attached Figure Description

[0038] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0039] Figure 1 This is a flowchart illustrating the preparation process of a standard Sophora japonica decoction according to the present invention.

[0040] Figure 2 For comparison feature maps. Detailed Implementation

[0041] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0042] Please see Figure 1 As shown, the present invention relates to the preparation of a standard decoction of Sophora japonica fruit, comprising the following steps:

[0043] Step S1: Weigh out the Sophora japonica fruit slices, place them in a clay pot, and decoct twice;

[0044] Step S2: Add 8 times the amount of water, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes.

[0045] Step S3: Add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 20 minutes;

[0046] Step S4: Filter the solution through a 200-mesh sieve, cool the filtrate to room temperature in a cold water bath, and combine the two filtrates;

[0047] Step S5: Cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate them under vacuum at 50°C;

[0048] Step S6: Concentrate to a fluid extract with a weight ratio of approximately 1:1 between the medicinal material and the fluid extract;

[0049] Step S7: Collect the concentrated solution and freeze-dry it to obtain the sample;

[0050] Step S8: Calculate and measure the indicators of traditional Chinese medicine in the decoction; after vacuum concentration of the Sophora japonica water extract, freeze-dry the concentrate until dry, collect the dried extract powder and weigh it, and calculate the extract yield; the formula for extract yield is as follows:

[0051]

[0052] This invention relates to a quality testing method for a standard decoction of Sophora japonica fruit, comprising the following steps:

[0053] Step A, Preparation of the reference solution: Take an appropriate amount of succinic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 0.1 mg per 1 ml.

[0054] Step B: Take an appropriate amount of this product, grind it into a fine powder, take about 0.2g, weigh it accurately, place it in a stoppered conical flask, accurately add 50ml of 50% methanol, weigh it, sonicate for 20 minutes, cool it, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, and take the filtrate to obtain the product.

[0055] Step C, Preparation of reference solution: Take 0.5g of Sophora japonica pod reference material, place it in a stoppered conical flask, add 50ml of 50% methanol, seal tightly, sonicate for 20 minutes, cool, shake well, filter, and take the filtrate as the reference solution of the reference material; separately take an appropriate amount of rutin reference standard, accurately weigh it, add methanol to prepare a solution containing 0.2mg per 1ml, as the reference solution of the reference standard;

[0056] Step D, Setup: Inject 10 μl each of the obtained reference solution, test solution, and standard solution into the liquid chromatograph for detection;

[0057] The conditions used for chromatographic detection in liquid chromatography include:

[0058] Chromatographic column: The column is packed with octadecylsilane-bonded silica gel, with a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm;

[0059] Column temperature: 30℃;

[0060] Mobile phase: Acetonitrile as mobile phase A, and 0.1% phosphoric acid as mobile phase B;

[0061] Mobile phase flow rate: 1.2 mL per minute;

[0062] Detection wavelength: 282nm;

[0063] The theoretical plate number, calculated based on the rutin peak, should be no less than 3000;

[0064] The mobile phase is eluted using a gradient program as follows:

[0065] From 0 to 12 min, the mobile phase A increased from 5% to 20%, while the mobile phase B decreased from 95% to 80%.

[0066] From 12 to 25 minutes, the concentration of mobile phase A increased from 20% to 22%, while the concentration of mobile phase B decreased from 80% to 78%.

[0067] Over 25–40 minutes, the mobile phase A concentration increased from 22% to 50%, while the mobile phase B concentration decreased from 78% to 50%.

[0068] The chromatogram of the test sample includes nine characteristic peaks, and the relative retention times of these peaks should correspond to those of the nine characteristic peaks in the chromatogram of the reference medicinal material. The peak corresponding to the rutin reference material is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 is 0.353, peak 2 is 0.679, peak 3 is 0.844, peak 4 is 0.945, peak 6 is 1.165, peak 7 is 1.220, peak 8 is 1.360, and peak 9 is 1.405.

[0069] Example 1

[0070] The preparation method of the standard decoction of Sophora japonica fruit is as follows: Weigh an appropriate amount of Sophora japonica fruit slices, place them in a clay pot, and decoct twice. For the first decoction, add 8 times the amount of water, soak for 30 minutes, bring to a boil over high heat, and then simmer over low heat for 30 minutes. For the second decoction, add 6 times the amount of water, bring to a boil over high heat, and then simmer over low heat for 20 minutes. Filter the decoction through a 200-mesh filter, and quickly cool the filtrate to room temperature in a water bath. Mix the two filtrates and concentrate them under vacuum at a temperature of 50°C until the weight ratio of the medicinal material to the fluid extract is approximately 1:1. Collect the concentrated liquid and freeze-dry it to obtain the sample.

[0071] After vacuum concentration of the aqueous extract of Sophora japonica fruit, the concentrate was freeze-dried to dryness. The dried extract powder was collected, weighed, and the yield was calculated. The formula for the yield is as follows:

[0072]

[0073] Ointment yield range: 17.5% to 28.0%.

[0074] The extractives were determined by hot extraction using ethanol as the solvent, according to the method for determining alcohol-soluble extractives (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201). The extractives range of the standard decoction was 40.2%–58.6%.

[0075] Content determination:

[0076] Sophoroside was determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020).

[0077] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); methanol-acetonitrile-0.07% phosphoric acid solution (12:20:68) was used as the mobile phase; the detection wavelength was 260 nm. The theoretical plate number, calculated based on the sophoroside peak, should be no less than 3000.

[0078] Preparation of reference solution: Take an appropriate amount of sophoroside reference standard, accurately weigh it, dissolve it in methanol (heat if necessary) to prepare a solution containing 25 μg per ml.

[0079] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 0.2g, weigh it accurately, place it in a stoppered conical flask, accurately add 50ml of 50% methanol, weigh it, sonicate it (power 600W, frequency 40kHz) for 20 minutes, cool it, weigh it again, make up the weight loss with 50% methanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0080] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0081]

[0082] This product contains 27.1mg to 66.8mg of juglone per 1g.

[0083] The transfer rate of succinic acid content in the decoction of medicinal materials ranged from 12.9% to 32.1%.

[0084] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0085] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile mobile phase A and 0.1% phosphoric acid solution mobile phase B were used for gradient elution according to the specifications in the table below; the column temperature was 30℃; the detection wavelength was 282 nm. The theoretical plate number, calculated based on the rutin peak, should not be less than 3000.

[0086] 30℃; detection wavelength 282nm. The theoretical plate number, calculated based on the rutin peak, should be no less than 3000.

[0087]

[0088] Preparation of reference solution: Take 0.5g of Sophora japonica pod reference material, place it in a stoppered conical flask, add 50ml of 50% methanol, stopper tightly, sonicate (power 600W, frequency 40kHz) for 20 minutes, cool, shake well, filter, and take the filtrate as the reference solution of the reference material; separately take an appropriate amount of rutin reference standard, accurately weigh it, add methanol to prepare a solution containing 0.2mg per ml, as the reference solution of the reference standard.

[0089] Preparation of test solution: Take an appropriate amount of this product, grind it into a fine powder, and take about 0.1g to prepare the test solution in the same way as the preparation method of the reference solution of the reference medicinal material.

[0090] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0091] The chromatogram of the test sample should show 9 characteristic peaks, and the retention times should correspond to the 9 characteristic peaks in the chromatogram of the reference medicinal material. The peak corresponding to the rutin reference peak is the S peak. Calculate the relative retention times of each characteristic peak and the S peak. The relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.353 (peak 1), 0.679 (peak 2), 0.844 (peak 3), 0.945 (peak 4), 1.165 (peak 6), 1.220 (peak 7), 1.360 (peak 8), and 1.405 (peak 9).

[0092] like Figure 2 As shown, peak 5 (S): rutin; peak 9: rutin; each milligram of the test sample is approximately equivalent to 0.018 grams of medicinal slices.

[0093] Example 2

[0094] According to the "Technical Requirements for Quality Control and Standardization of Traditional Chinese Medicine Formula Granules," the samples used in the research should be representative, covering the main producing areas or authentic production areas of the medicinal materials to be harvested, with no fewer than three batches from each producing area. Sophora japonica fruit is a cultivated product, distributed throughout my country. It is mainly produced in Guangzhou and Henan provinces, where its cultivation has reached a large scale and is the main source of commercial Sophora japonica fruit. Based on a comprehensive evaluation of raw material quality, sustainable resource supply, and other factors, the preliminary raw material producing areas for large-scale production were determined to be Panyu District of Guangzhou, Nanzhao County of Nanyang City in Henan Province, Biyang County of Zhumadian City in Henan Province, Tongbai County of Nanyang City in Henan Province, and Ruyang County of Luoyang City in Henan Province. Ultimately, 15 qualified batches were selected for the study of standard decoctions. The transfer rate of the 15 batches of Sophora japonica fruit standard decoctions is shown in the table below:

[0095]

[0096]

[0097] Based on the content of 15 batches of Sophora japonica fruit slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 22.5%.

[0098] The range calculated using the mean plus or minus 3 times the SD is 12.9% to 32.1%.

[0099] The range calculated using 70% to 130% of the mean is 15.7% to 29.2%.

[0100] Based on the actual range of 16.5% to 26.3%;

[0101] It is recommended to take the average plus or minus 3 times the SD, i.e., 12.9% to 32.1%, as the upper and lower limits of the standard decoction transfer rate.

[0102] Example 3

[0103] Based on the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules", and combined with data from 15 batches of Sophora japonica fruit standard decoction, the content limits and upper and lower limits of the standard decoction were calculated.

[0104] The range calculated using the mean plus or minus 3 times the SD is: 27.1 mg to 75.6 mg;

[0105] The range calculated using 70%–130% of the mean is 35.9 mg–66.8 mg.

[0106] Based on the actual range of 36.1mg to 63.9mg, it is recommended to use the mean of 70% to 130% and the mean plus or minus 3 times the SD value, i.e., 27.1mg to 66.8mg, as the upper and lower limits of the standard decoction content.

[0107] Based on the content of 15 batches of Sophora japonica fruit slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 22.5%.

[0108] The range calculated using the mean plus or minus 3 times the SD is 12.9% to 32.1%.

[0109] The range calculated using 70% to 130% of the mean is 15.7% to 29.2%.

[0110] Based on the actual range of 16.5% to 26.3%;

[0111] It is recommended to take the average plus or minus 3 times the SD, i.e., 12.9% to 32.1%, as the upper and lower limits of the standard decoction transfer rate.

[0112] Establishing a comparative characteristic spectrum: Based on the data analysis results, nine common peaks were selected as characteristic peaks. The generated standard spectrum is shown below. Figure 2 Using succinic acid as a reference peak, the relative retention times of characteristic peaks in the chromatogram were calculated.

[0113] By comparing the chromatogram with that of the reference standard, it was found that among the nine characteristic peaks in the characteristic chromatogram of the Sophora japonica standard decoction, peak 5 (S) is rutin. According to the national drug standard YBZ-PFKL-2021062 for Sophora japonica formula granules, peak 5 is rutin and peak 9 is sophoroside.

[0114] The relative retention times of the five batches of Sophora japonica fruit standard decoction are shown in the table below:

[0115]

[0116] Retention time of characteristic spectrum of medicinal slices

[0117]

[0118]

[0119] The results showed that the characteristic chromatograms of the five batches of Sophora japonica standard decoction samples all exhibited chromatographic peaks corresponding to the retention times of the nine main characteristic peaks in the reference medicinal material. The relative retention time RSDs of peaks 1-4 and peaks 6-9 were less than 2%, indicating that the elution times of each peak were consistent across batches. (Note: The batch number of the standard decoction was selected to correspond to the batch of medicinal slices to prove that the prepared sample had almost no effect on the characteristic chromatogram of the medicinal slices.)

[0120] The national standard YBZ-PFKL-2021062, "Sophora japonica granules," specifies the relative retention times of peak 5 (rutin), peaks 1-4, peaks 6-9, and peak 5 (S). Based on the results of 15 batches of Sophora japonica standard decoctions and the retention time data from repeatability and precision tests in the methodology (see Tables 4-12 and 4-14 below), it is known that the retention times of peaks 1-4 and 6-9 are stable, and the relative peak drift of peaks 1-4 and 6-9 is very small, generally within ±7%. Therefore, it is stipulated that the chromatogram of the test sample should show 9 characteristic peaks, corresponding to the retention times of the 9 characteristic peaks in the chromatogram of the reference medicinal material. The peak corresponding to the rutin reference peak is the S peak. The relative retention times of each characteristic peak and the S peak should be calculated, and their relative retention times should be within ±10% of the specified value. The specified values ​​are: 0.353 (peak 1), 0.679 (peak 2), 0.844 (peak 3), 0.945 (peak 4), 1.165 (peak 6), 1.220 (peak 7), 1.360 (peak 8), and 1.405 (peak 9).

[0121] According to the "Leachate Content" section of the "Sophora japonica Fruit Formula Granules" standard YBZ-PFKL-2021141, the finished granule product is required to include a "Leachate Content" test item. Referring to the method for determining the leachate content of the finished granule product, the leachate content test results for 15 batches of Sophora japonica fruit are shown in the table below:

[0122]

[0123]

[0124] The average value of the alcohol-soluble extract from 15 batches of standard decoction was 49.4%.

[0125] The range calculated using the mean plus or minus 3 times the SD is 40.2% to 58.6%.

[0126] The range calculated using 70% to 130% of the mean is 34.6% to 64.2%.

[0127] Based on the actual range, it is 46.7% to 55.7%;

[0128] A range of 40.2% to 58.6% is recommended.

[0129] It is worth noting that, regarding the upper and lower limits of the standard decoction extract, the units included in the above system embodiments are only divided according to functional logic, but are not limited to the above division, as long as the corresponding function can be achieved; in addition, the specific names of each functional unit are only for easy differentiation and are not used to limit the scope of protection of the present invention.

[0130] Furthermore, those skilled in the art will understand that all or part of the steps in the methods of the above embodiments can be implemented by a program instructing related hardware, and the corresponding program can be stored in a computer-readable storage medium.

[0131] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to the specific implementations described. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. A method for preparing a standard decoction of Sophora japonica fruit, characterized in that, Includes the following steps: Step S1: Weigh out the Sophora japonica fruit slices, place them in a clay pot, and decoct twice; Step S2: Add 8 times the amount of water, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes. Step S3: Add 6 times the amount of water, bring to a boil over high heat, then simmer over low heat for 20 minutes; Step S4: Filter the solution through a 200-mesh sieve, cool the filtrate to room temperature in a cold water bath, and combine the two filtrates; Step S5: Cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate them under vacuum at 50°C; Step S6: Concentrate to a fluid extract with a weight ratio of approximately 1:1 between the medicinal material and the fluid extract; Step S7: Collect the concentrated solution and freeze-dry it to obtain the sample; Step S8: Calculate and measure the indicators of Chinese medicine in the decoction.

2. The method for preparing a standard decoction of Sophora japonica fruit according to claim 1, characterized in that, In step S8, the aqueous extract of Sophora japonica is concentrated under vacuum, and the concentrate is freeze-dried to dryness. The dried extract powder is collected and weighed, and the yield is calculated. The formula for the yield is as follows:

3. A quality testing method for a standard decoction of Sophora japonica fruit, characterized in that, Includes the following steps: Step A, Preparation of the reference solution: Take an appropriate amount of succinic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 0.1 mg per 1 ml. Step B: Take an appropriate amount of this product, grind it into a fine powder, take about 0.2g, weigh it accurately, place it in a stoppered conical flask, accurately add 50ml of 50% methanol, weigh it, sonicate for 20 minutes, cool it, weigh it again, make up the lost weight with 50% methanol, shake it well, filter it, and take the filtrate to obtain the product. Step C, Preparation of reference solution: Take 0.5g of Sophora japonica pod reference material, place it in a stoppered conical flask, add 50ml of 50% methanol, seal tightly, sonicate for 20 minutes, cool, shake well, filter, and take the filtrate as the reference solution of the reference material; separately take an appropriate amount of rutin reference standard, accurately weigh it, add methanol to prepare a solution containing 0.2mg per 1ml, as the reference solution of the reference standard; Step D, Setup: Inject 10 μl each of the obtained reference solution, test solution, and standard solution into the liquid chromatograph for detection; The conditions used for chromatographic detection in the liquid chromatograph include: Chromatographic column: The column is packed with octadecylsilane-bonded silica gel, with a length of 250 mm, an inner diameter of 4.6 mm, and a particle size of 5 μm; Column temperature: 30℃; Mobile phase: Acetonitrile as mobile phase A, and 0.1% phosphoric acid as mobile phase B; Mobile phase flow rate: 1.2 mL per minute; Detection wavelength: 282nm; The theoretical plate number, calculated based on the rutin peak, should be no less than 3000; The mobile phase is eluted using a gradient program as follows: From 0 to 12 min, the mobile phase A increased from 5% to 20%, while the mobile phase B decreased from 95% to 80%. From 12 to 25 minutes, the concentration of mobile phase A increased from 20% to 22%, while the concentration of mobile phase B decreased from 80% to 78%. Over 25–40 minutes, the mobile phase A concentration increased from 22% to 50%, while the mobile phase B concentration decreased from 78% to 50%.

4. The quality testing method for a standard decoction of Sophora japonica fruit according to claim 1, characterized in that, The chromatogram of the test sample includes nine characteristic peaks, and the relative retention times of these peaks should correspond to those of the nine characteristic peaks in the chromatogram of the reference medicinal material. The peak corresponding to the rutin reference material is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 is 0.353, peak 2 is 0.679, peak 3 is 0.844, peak 4 is 0.945, peak 6 is 1.165, peak 7 is 1.220, peak 8 is 1.360, and peak 9 is 1.405.

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