Preparation method and quality detection method of vinegar rhizoma corydalis standard decoction

Abstract

The invention discloses a preparation method and a quality detection method of a vinegar rhizoma corydalis standard decoction, and relates to the technical field of quality research of traditional Chinese medicinal materials. The preparation method comprises the following steps: weighing a proper amount of vinegar-processed rhizoma corydalis decoction pieces, putting into a marmite, decocting twice, adding 9 times of water for soaking for 30 minutes for the first time, boiling with strong fire, and decocting with slow fire for 30 minutes; 7 times of water is added for the second time, the mixture is boiled with strong fire and then decocted with slow fire for 20 min, the liquid medicine is filtered with 200 meshes, the filtrate is cooled to the room temperature in a cold water bath, the two filtrates are combined, vacuum concentration is conducted at the temperature of 50 DEG C, vacuum concentration is conducted at the absolute vacuum degree of 30-40 mbar until the weight ratio of the medicinal materials to the fluid extract is about 1: 1, concentrated liquid is collected, and freeze drying is conducted to obtain a sample. According to the method, key quality indexes such as the main content, the characteristic chromatogram and the extract of the vinegar rhizoma corydalis after the decoction is prepared are measured, the transfer rate is investigated, the component transfer condition from 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 vinegar-processed Corydalis yanhusuo and its quality testing method. Background Technology

[0002] Modern research shows that Corydalis rhizome contains various components, such as alkaloids (isoquinoline alkaloids, apophene alkaloids, protoberberine alkaloids, and other alkaloids) and non-alkaloid compounds (in addition to basic non-alkaloid compounds such as water, protein, mucilage, and volatile oil, acetic corydalis rhizome also contains nearly 10 kinds of non-alkaloid compounds). Acetic corydalis rhizome has effects on the central nervous system (sedation, anti-anxiety, hypnosis, analgesia), the digestive system (treatment and adjuvant treatment of gastric ulcers), the cardiovascular system (good protective effect against myocardial ischemia), the pituitary-adrenal cortex system, and anti-tumor activity, among other pharmacological activities. Among them, tetrahydropalmatine (dl-THP), also known as tetrahydropalmatine, has a weaker analgesic effect than pethidine and morphine, but is less addictive and stronger than general antipyretic analgesics. It is effective for chronic, persistent dull pain, but less effective for post-traumatic or post-operative pain, or pain associated with advanced cancer. Ethylenediol hydrochloride is the main active substance in acetic corydalis, inducing hypotension and bradycardia in rats by antagonizing striatal dopamine D2 receptors. It has significant therapeutic effects on pain caused by the gastrointestinal and hepatobiliary systems, and can also relieve labor pains, headaches, and concussion headaches. It is also used to eliminate mental tension such as irritability and anxiety, and to treat neurasthenia-related insomnia. The 2020 edition of the Chinese Pharmacopoeia lists acethylenediol as an indicator component for quality control of acetic corydalis. Combined with the "Quality Standard for Ethylened Corydalis Formula Granules" YBZ-PFKL-2021133, which specifies acethylenediol as a content quality control component, this study selected acethylenediol as the quality control indicator component for the standard acetic corydalis 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 the fingerprint chromatograms of vinegar-processed Corydalis rhizome mostly employ conventional HPLC methods, and these methods only target the material basis of the original medicinal material. The indicator 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 herbal decoctions. Summary of the Invention

[0005] The purpose of this invention is to provide a method for preparing a standard decoction of vinegar-processed Corydalis and for its quality testing. By measuring key quality indicators such as the main content, characteristic spectrum, and extract of vinegar-processed Corydalis 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 vinegar-processed Corydalis 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 Corydalis yanhusuo in vinegar, comprising the following steps:

[0008] Step S1: Weigh out the vinegar-processed Corydalis slices, place them in a clay pot, and decoct twice;

[0009] Step S2: Add 9 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 7 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 a temperature of 50°C and an absolute vacuum of 30-40 mbar.

[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 Corydalis yanhusuo is concentrated under vacuum, and then 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 Corydalis yanhusuo in vinegar, comprising the following steps:

[0019] Step A, Preparation of the reference solution: Take an appropriate amount of acetocopherol reference standard, accurately weigh it, dissolve it in methanol to prepare a solution containing 35 μg per ml;

[0020] Step B, 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 25ml of dilute ethanol, weigh it, sonicate for 30 minutes, cool it, weigh it again, make up the lost weight with dilute ethanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0021] Step C, Preparation of reference solution: Take appropriate amounts of protopine reference standard, palmatine hydrochloride reference standard, berberine hydrochloride reference standard and acetocopherol reference standard, accurately weigh them, add methanol to prepare a mixed solution containing 10 μg of protopine, 10 μg of palmatine hydrochloride, 5 μg of berberine hydrochloride and 100 μg of acetocopherol per 1 ml, as the reference solution;

[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: 1.0 mL per minute;

[0028] Detection wavelength: 280nm;

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

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

[0031] From 0 to 15 min, the mobile phase A increased from 10 to 17%, while the mobile phase B decreased from 90 to 83%.

[0032] From 15 to 65 minutes, the concentration of mobile phase A increased from 17 to 30%, while the concentration of mobile phase B decreased from 83 to 70%.

[0033] From 65 to 85 minutes, the concentration of mobile phase A increased from 30% to 55%, while the concentration of mobile phase B decreased from 70% to 45%.

[0034] From 85 to 105 min, the mobile phase A increased from 55% to 80%, while the mobile phase B decreased from 45% to 20%.

[0035] From 105 to 115 min, the mobile phase A increased from 80% to 85%, while the mobile phase B decreased from 20% to 15%.

[0036] At 115–116 min, the concentration of mobile phase A decreased from 85% to 10%, while the concentration of mobile phase B increased from 15% to 90%.

[0037] Between 116 and 130 minutes, the mobile phase A increased from 29% to 30%, while the mobile phase B reached 90%.

[0038] As a preferred technical solution, the chromatogram of the test sample includes four characteristic peaks, which should correspond to the relative retention times of the four characteristic peaks in the chromatogram of the reference medicinal material; wherein, the peak corresponding to the reference material of acetocopherol is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 not less than 0.398, peak 2 not less than 1.107, and peak 3 not less than 0.351.

[0039] The present invention has the following beneficial effects:

[0040] This invention measures key quality indicators such as the main content, characteristic spectrum, and extract of vinegar-processed Corydalis yanhusuo after decoction preparation, investigates the transfer rate, judges the range of transfer rate of the main content of the medicinal material, and detects the effectiveness of the decoction, thereby improving the technical level and scientific content of traditional Chinese medicine quality control.

[0041] 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

[0042] 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.

[0043] Figure 1 This is a flowchart illustrating the preparation process of a standard decoction of Corydalis yanhusuo in vinegar according to the present invention.

[0044] Figure 2 For comparison feature maps. Detailed Implementation

[0045] 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.

[0046] Please see Figure 1 As shown, this invention relates to the preparation of a standard decoction of Corydalis yanhusuo in vinegar, comprising the following steps:

[0047] Step S1: Weigh out the vinegar-processed Corydalis slices, place them in a clay pot, and decoct twice;

[0048] Step S2: Add 9 times the amount of water, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes;

[0049] Step S3: Add 7 times the amount of water, bring to a boil over high heat, then simmer over low heat for 20 minutes;

[0050] 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;

[0051] Step S5: Cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate them under vacuum at a temperature of 50°C and an absolute vacuum of 30-40 mbar.

[0052] Step S6: Concentrate to a fluid extract with a weight ratio of approximately 1:1 between the medicinal material and the fluid extract;

[0053] Step S7: Collect the concentrated solution and freeze-dry it to obtain the sample;

[0054] Step S8: Calculate and measure the indicators of traditional Chinese medicine in the decoction; after vacuum concentration of the vinegar-processed Corydalis yanhusuo aqueous 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:

[0055]

[0056] This invention relates to a quality testing method for a standard decoction of Corydalis yanhusuo in vinegar, comprising the following steps:

[0057] Step A, Preparation of the reference solution: Take an appropriate amount of acetocopherol reference standard, accurately weigh it, dissolve it in methanol to prepare a solution containing 35 μg per ml;

[0058] Step B, 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 25ml of dilute ethanol, weigh it, sonicate for 30 minutes, cool it, weigh it again, make up the lost weight with dilute ethanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0059] Step C, Preparation of reference solution: Take appropriate amounts of protopine reference standard, palmatine hydrochloride reference standard, berberine hydrochloride reference standard and acetocopherol reference standard, accurately weigh them, add methanol to prepare a mixed solution containing 10 μg of protopine, 10 μg of palmatine hydrochloride, 5 μg of berberine hydrochloride and 100 μg of acetocopherol per 1 ml, as the reference solution;

[0060] Step D, Setup: Inject 10 μl each of the obtained reference solution, test solution, and standard solution into the liquid chromatograph for detection;

[0061] The conditions used for chromatographic detection in liquid chromatography include:

[0062] 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;

[0063] Column temperature: 30℃;

[0064] Mobile phase: Acetonitrile as mobile phase A, and 0.1% phosphoric acid as mobile phase B;

[0065] Mobile phase flow rate: 1.0 mL per minute;

[0066] Detection wavelength: 280nm;

[0067] The theoretical plate number, calculated based on the peak of acetocopherol acetate, should be no less than 3000;

[0068] The mobile phase is eluted using a gradient program as follows:

[0069] From 0 to 15 min, the mobile phase A increased from 10 to 17%, while the mobile phase B decreased from 90 to 83%.

[0070] From 15 to 65 minutes, the concentration of mobile phase A increased from 17 to 30%, while the concentration of mobile phase B decreased from 83 to 70%.

[0071] From 65 to 85 minutes, the concentration of mobile phase A increased from 30% to 55%, while the concentration of mobile phase B decreased from 70% to 45%.

[0072] From 85 to 105 min, the mobile phase A increased from 55% to 80%, while the mobile phase B decreased from 45% to 20%.

[0073] From 105 to 115 min, the mobile phase A increased from 80% to 85%, while the mobile phase B decreased from 20% to 15%.

[0074] At 115–116 min, the concentration of mobile phase A decreased from 85% to 10%, while the concentration of mobile phase B increased from 15% to 90%.

[0075] Between 116 and 130 minutes, the mobile phase A increased from 29% to 30%, while the mobile phase B reached 90%.

[0076] The chromatogram of the test sample includes four characteristic peaks, and the relative retention times of the four characteristic peaks in the chromatogram of the reference medicinal material should correspond to those of the reference material. Among them, the peak corresponding to the reference material of acetocopherol is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 not less than 0.398, peak 2 not less than 1.107, and peak 3 not less than 0.351.

[0077] Example 1

[0078] The preparation method of the standard decoction of vinegar-processed Corydalis is as follows: Weigh an appropriate amount of vinegar-processed Corydalis slices, place them in a clay pot, and decoct twice. For the first decoction, add 9 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 7 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, cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate under vacuum at a temperature of 50℃ and an absolute vacuum of 30-40 mbar 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.

[0079] After vacuum concentration of the aqueous extract of Corydalis yanhusuo, 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:

[0080]

[0081] Ointment yield range: 14.5% to 21.7%.

[0082] 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 12.1%–21.5%.

[0083] Content determination:

[0084] Ethyroxine was determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020).

[0085] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-0.1% phosphoric acid solution (adjusted to pH 6.0 with triethylamine) (55:45) was used as the mobile phase; the flow rate was 1.0 mL / min; the column temperature was 30℃; and the detection wavelength was 280 nm. The theoretical plate number, calculated based on the acetophenone peak, should be no less than 3000.

[0086] Preparation of reference solution: Take an appropriate amount of acetocopherol reference standard, accurately weigh it, dissolve it in methanol (heat if necessary) to prepare a solution containing 30 μg per ml.

[0087] 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 25ml of dilute ethanol, weigh it, sonicate it (power 250W, frequency 40kHz) for 30 minutes, cool it, weigh it again, make up the weight loss with dilute ethanol, shake it well, filter it, and take the filtrate to obtain the test solution.

[0088] Assay: Accurately pipette 10 μl each of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0089]

[0090] This product contains 1.1mg to 5.2mg of corydaline per 1g.

[0091] The transfer rate of corydaline content in the decoction of medicinal materials ranged from 29.6% to 54.9%.

[0092] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0093] 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 was used as mobile phase A, and 0.1% phosphoric acid solution (adjusted to pH 6.0 with triethylamine) was used as mobile phase B, with gradient elution as specified in the table below; the flow rate was 1.0 mL / min; the column temperature was 30℃; and the detection wavelength was 280 nm. The theoretical plate number, calculated based on the acetophenone peak, should be no less than 3000.

[0094]

[0095]

[0096] Preparation of reference solution: Take appropriate amounts of protopine reference standard, palmatine hydrochloride reference standard, berberine hydrochloride reference standard and acetocopherol reference standard, accurately weigh them, add methanol to prepare a mixed solution containing 10 μg of protopine, 10 μg of palmatine hydrochloride, 5 μg of berberine hydrochloride and 100 μg of acetocopherol per 1 ml, as the reference solution.

[0097] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take 0.3g, place it in a stoppered conical flask, add 100ml of 30% methanol, stopper tightly, sonicate (power 200W, frequency 50kHz) for 30 minutes, take it out, cool it, filter it, and take the filtrate to obtain the test solution.

[0098] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0099] The chromatogram of the test sample should show four characteristic peaks, each corresponding to the retention time of the corresponding reference peak. The peak corresponding to the reference peak of acetocopherol is designated as the S peak. The relative peak areas of each characteristic peak and the S peak should be calculated, and these relative peak areas should be within the specified ranges: not less than 0.312 (peak 1), not less than 0.512 (peak 2), and not less than 0.141 (peak 3).

[0100] like Figure 2 Peak 1: Protopine; Peak 2: Palmatine hydrochloride; Peak 3: Berberine hydrochloride; Peak 4 (S): Ethylenediol, each milligram of the test sample is approximately equivalent to 0.044 grams of medicinal slices.

[0101] Example 2

[0102] 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. Vinegar-processed Corydalis Rhizome 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 vinegar-processed Corydalis Rhizome. 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, Henan, Biyang County of Zhumadian, Henan, Tongbai County of Nanyang, Henan, and Ruyang County of Luoyang, Henan. Ultimately, 15 qualified batches were selected for research on standard decoctions.

[0103] The transfer rates of 15 batches of vinegar-processed Corydalis yanhusuo standard decoction are shown in the table below:

[0104]

[0105]

[0106] Based on the content of 15 batches of vinegar-processed Corydalis rhizome slices, the content of standard decoction, and the yield of extract, the average transfer rate of the standard decoction was calculated to be 44.5%.

[0107] The range calculated using the mean plus or minus 3 times the SD is 34.6% to 54.3%.

[0108] The range calculated using 70% to 130% of the mean is 31.1% to 57.8%.

[0109] Based on the actual range, it is 39.3% to 49.2%;

[0110] It is recommended to take the average value of 70% to 130%, or 31.1% to 57.8%, as the upper and lower limits of the standard decoction transfer rate.

[0111] Example 3

[0112] 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 vinegar-processed Corydalis Rhizome standard decoction, the content limits and upper and lower limits of the standard decoction were calculated.

[0113] The range calculated using the mean plus or minus 3 times the SD is: 1.2 mg to 5.4 mg;

[0114] The range calculated using 70%–130% of the mean is 2.3 mg–4.3 mg.

[0115] Based on the actual range of 2.5mg to 5.2mg, it is recommended to use the mean plus or minus 3 times the SD, i.e., 1.2mg to 5.4mg, as the upper and lower limits of the standard decoction content.

[0116] Based on the content of 15 batches of vinegar-processed Corydalis rhizome slices, the content of standard decoction, and the yield of extract, the average transfer rate of the standard decoction was calculated to be 44.5%.

[0117] The range calculated using the mean plus or minus 3 times the SD is 34.6% to 54.3%.

[0118] The range calculated using 70% to 130% of the mean is 31.1% to 57.8%.

[0119] Based on the actual range, it is 39.3% to 49.2%;

[0120] It is recommended to take the average value of 70% to 130%, or 31.1% to 57.8%, as the upper and lower limits of the standard decoction transfer rate.

[0121] The standard decoction of Corydalis yanhusuo with vinegar, batch number BT(YHS)220401, was selected as the reference chromatogram to establish a control characteristic chromatogram. Based on the data analysis results, four common peaks were finally selected as characteristic peaks. The generated standard chromatogram is shown below. Figure 2 Using acetocopherol as a reference peak, the relative retention times of the characteristic peaks in the chromatogram were calculated.

[0122] By comparing the chromatogram with that of the reference standard, it can be seen that among the four characteristic peaks in the characteristic chromatogram of the vinegar-corydalis standard decoction, peak 4(S) is corydalis ethyl acetate. This can be confirmed by referring to the characteristic chromatogram of the national drug standard YBZ-PFKL-2021133 for "Vinegar-corydalis Formula Granules".

[0123]

[0124] The results showed that the characteristic chromatograms of the 15 batches of vinegar-processed Corydalis standard decoction samples all showed chromatographic peaks corresponding to the retention times of the four main characteristic peaks in the control herb. The RSD of the retention times of peaks 1 to 4 was less than 2%, indicating that the elution times of each peak were consistent among the batches. (Note: The batch number of the standard decoction was selected to correspond to the batch of the medicinal slices to prove that the prepared sample had almost no effect on the characteristic chromatogram of the medicinal slices.)

[0125] The national standard YBZ-PFKL-2021031, "Quality Standard for Corydalis Rhizome Granules," stipulates that the retention times of the four characteristic peaks should correspond to the retention times of the reference peaks, and the relative peak areas of peaks 1-3 and peak 4 should also be specified. Based on the test results of six batches of Corydalis Rhizome standard decoctions and the retention time data from the repeatability and precision tests in the methodology, it can be seen that the retention times of the four characteristic peaks are stable and correspond to the retention times of the reference peaks. The average relative peak areas of peaks 1-3 and peak 4 are not less than 0.398 (peak 1), not less than 1.107 (peak 2), and not less than 0.351 (peak 3), respectively. Therefore, it is stipulated that the chromatogram of the test sample should show four characteristic peaks, and the retention times of each of the four peaks should correspond to the retention times of the corresponding reference peaks. The peak corresponding to the reference peak of corydalis ethyl is called the S peak. Calculate the relative peak area of ​​each characteristic peak and the S peak. The relative peak area should be within the specified range, which is: not less than 0.398 (peak 1), not less than 1.107 (peak 2), and not less than 0.351 (peak 3).

[0126] Using ethanol as solvent, the extractives of 15 batches of vinegar-processed Corydalis rhizome were determined by the hot extraction method under the alcohol-soluble extractives determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201). The results are shown in the table below:

[0127]

[0128]

[0129]

[0130] The average value of the alcohol-soluble extract from 15 batches of standard decoction was 16.8%.

[0131] The range calculated using the mean plus or minus 3 times the SD is 12.1% to 21.5%.

[0132] The range calculated using 70% to 130% of the mean is 11.8% to 21.8%.

[0133] Based on the actual range, it is 14.5% to 19.2%;

[0134] It is recommended to use the mean plus or minus 3 times the SD, i.e., 12.1% to 21.5%, as the upper and lower limits of the standard decoction extract.

[0135] It is worth noting that the various 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 functions 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.

[0136] 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.

[0137] 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 Corydalis yanhusuo in vinegar, characterized in that, Includes the following steps: Step S1: Weigh out the vinegar-processed Corydalis slices, place them in a clay pot, and decoct twice; Step S2: Add 9 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 7 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 a temperature of 50°C and an absolute vacuum of 30-40 mbar. 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 Corydalis yanhusuo in vinegar according to claim 1, characterized in that, In step S8, the aqueous extract of Corydalis yanhusuo 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:

3. A quality testing method for a standard decoction of Corydalis yanhusuo in vinegar, characterized in that, Includes the following steps: Step A, Preparation of the reference solution: Take an appropriate amount of acetocopherol reference standard, accurately weigh it, dissolve it in methanol to prepare a solution containing 35 μg per ml; Step B, 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 25ml of dilute ethanol, weigh it, sonicate for 30 minutes, cool it, weigh it again, make up the lost weight with dilute ethanol, shake it well, filter it, and take the filtrate to obtain the test solution. Step C, Preparation of reference solution: Take appropriate amounts of protopine reference standard, palmatine hydrochloride reference standard, berberine hydrochloride reference standard and acetocopherol reference standard, accurately weigh them, add methanol to prepare a mixed solution containing 10 μg of protopine, 10 μg of palmatine hydrochloride, 5 μg of berberine hydrochloride and 100 μg of acetocopherol per 1 ml, as the reference solution; 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: 1.0 mL per minute; Detection wavelength: 280nm; The theoretical plate number, calculated based on the peak of acetocopherol acetate, should be no less than 3000; The mobile phase is eluted using a gradient program as follows: From 0 to 15 min, the mobile phase A increased from 10 to 17%, while the mobile phase B decreased from 90 to 83%. From 15 to 65 minutes, the concentration of mobile phase A increased from 17 to 30%, while the concentration of mobile phase B decreased from 83 to 70%. From 65 to 85 minutes, the concentration of mobile phase A increased from 30% to 55%, while the concentration of mobile phase B decreased from 70% to 45%. From 85 to 105 min, the mobile phase A increased from 55% to 80%, while the mobile phase B decreased from 45% to 20%. From 105 to 115 min, the mobile phase A increased from 80% to 85%, while the mobile phase B decreased from 20% to 15%. At 115–116 min, the concentration of mobile phase A decreased from 85% to 10%, while the concentration of mobile phase B increased from 15% to 90%. Between 116 and 130 minutes, the mobile phase A increased from 29% to 30%, while the mobile phase B reached 90%.

4. The quality testing method for a standard decoction of Corydalis yanhusuo in vinegar according to claim 1, characterized in that, The chromatogram of the test sample includes four characteristic peaks, and the relative retention times of the four characteristic peaks in the chromatogram of the reference medicinal material should correspond to those of the reference material. Among them, the peak corresponding to the reference material of acetocopherol is the S peak, and the relative retention times of each characteristic peak are as follows: peak 1 not less than 0.398, peak 2 not less than 1.107, and peak 3 not less than 0.351.

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