Preparation process of standard pueraria thomsonii decoction

Abstract

The invention discloses a preparation process of a standard pueraria thomsonii decoction, and relates to the technical field of traditional Chinese medicine decoction preparation. The preparation method comprises the following steps: firstly, weighing a proper amount of pachyrhizua angulatus decoction pieces, putting the pachyrhizua angulatus decoction pieces into a marmite, decocting twice, adding 9 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; 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, filtrate is rapidly cooled to the room temperature in a water bath mode, the two filtrates are mixed, vacuum concentration is conducted at the temperature of 50 DEG C 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 present invention, the pachyrhizua angulatus decoction is adopted as the standard, the key quality indexes such as the main content, the characteristic spectrum, the extract and the like of the single traditional Chinese medicine variety after the decoction preparation are stipulated, the transfer rate is investigated, the transfer rate range of the main content of the pachyrhizua angulatus medicinal material is stipulated, and the effectiveness of the pachyrhizua angulatus decoction is ensured according to the standard preparation method;

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine decoction preparation technology, and in particular to the preparation process of kudzu root standard decoction. Background Technology

[0002] Kudzu root is a perennial vine belonging to the genus *Pueraria* of the legume family. It is a green food that can be used for both medicinal and culinary purposes. It is high in starch and contains inorganic elements such as iron and calcium, as well as essential amino acids. Kudzu root is used medicinally to relieve muscle tension and fever, promote body fluid production and quench thirst, clear the meridians and collaterals, and detoxify alcohol. It can also be used for external fever and headache, lowering blood lipids and blood sugar, relieving fever, and protecting the cardiovascular system. Traditional Chinese medicine decoctions are a widely used form of medication in clinical practice. Standardized decoctions are single-herb Chinese medicine decoctions prepared using standardized processes based on traditional Chinese medicine theory and clinical application, with reference to modern extraction methods. They are used for standardized clinical medication to ensure the accuracy of medication and the consistency of dosage.

[0003] Currently, there are no quality studies or standards for kudzu root decoctions, making it impossible to measure the quality and efficacy of the decoction after preparation. This also makes it impossible to provide a basis for setting quality standards for the final product of kudzu root decoctions, leaving room for improvement. Therefore, we propose a preparation process for a standard kudzu root decoction. Summary of the Invention

[0004] The purpose of this invention is to provide a preparation process for a standard kudzu root decoction to solve the problems mentioned in the background art.

[0005] This invention is achieved through the following technical solution:

[0006] This invention relates to a preparation process for a standard kudzu root decoction, comprising the following steps:

[0007] S1: Place the kudzu root slices in a clay pot, add 9 times the amount of water, and soak for 30 minutes. Bring to a boil over high heat, then simmer over low heat for 30 minutes to obtain one batch of medicinal liquid. Pour out the first batch of medicinal liquid.

[0008] S2: Add 7 times the amount of water to the clay pot again, bring to a boil over high heat, then simmer over low heat for 20 minutes to obtain a second decoction. Pour out the second decoction.

[0009] S3: Filter the primary and secondary medicinal solutions obtained from S1 and S2 through a 200-mesh filter, cool the filtrate to room temperature in a cold water bath, and then combine the two filtrates to obtain the kudzu root medicinal solution;

[0010] S4: The obtained kudzu root extract was concentrated under vacuum at a temperature of 50°C until the weight ratio of the medicinal material to the fluid extract was 1:1.

[0011] S5: Based on S4 above, the concentrated liquid is collected and freeze-dried to obtain the standard kudzu root decoction.

[0012] Concentration and freeze-drying are used to measure the yield and extract indicators of kudzu root in decoctions.

[0013] In this invention, standard decoctions serve as a "bridge" connecting traditional Chinese medicine decoction pieces and modern Chinese medicine preparations. They provide a reference for controlling the quality of Chinese medicine end products, a tool for standardizing different forms of Chinese medicine administration to ensure the uniformity of quality and the consistency of efficacy, and a reference for evaluating the consistency of product quality from different manufacturers. Therefore, the research on the quality standards of standard decoctions of Chinese medicine decoction pieces will provide a foundation for the formulation of quality standards for all end products derived from decoctions of Chinese medicine decoction pieces.

[0014] The present invention has the following beneficial effects:

[0015] The preparation process of the standard kudzu root decoction of the present invention uses objective quality standards to measure the component transfer of kudzu root slices into decoction, and ensures the quality and effectiveness of kudzu root decoction.

[0016] The "standard" in the kudzu root standard decoction of this invention mainly covers the representativeness of the raw materials, the consistency of the preparation process with the traditional method, and the rigor of quality control, which basically ensures the stability and uniformity of the efficacy and quality of the kudzu root prescription. In addition, the kudzu root standard decoction has not undergone a drying process, thus maintaining the consistency with the traditional decoction in clinical practice to the greatest extent.

[0017] 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

[0018] 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.

[0019] Figure 1 This is a flowchart illustrating the preparation process of the standard kudzu root decoction of the present invention.

[0020] Figure 2 This is a characteristic peak comparison spectrum of the present invention;

[0021] Figure 3 This is a comparative characteristic chromatogram of the standard kudzu root decoction of the present invention. Detailed Implementation

[0022] 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.

[0023] Please refer to Figure 1 As shown, this invention relates to a preparation process for a standard kudzu root decoction, the method comprising:

[0024] Weigh an appropriate amount of kudzu root 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, 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, then simmer over low heat for 20 minutes. Filter the decoction through a 200-mesh filter, and rapidly cool the filtrate to room temperature in a water bath. Mix the two filtrates and concentrate them under vacuum at 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.

[0025] Extract yield: After vacuum concentration of the kudzu root aqueous extract, the concentrate is freeze-dried to dryness. The dried extract powder is collected and weighed, and the extract yield is calculated. The formula for extract yield is as follows:

[0026]

[0027] Ointment yield range: 18%–31%.

[0028] Extractives: Determined by hot extraction method under the alcohol-soluble extractives determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201) using ethanol as solvent. The extractives of the standard decoction range from 36.3% to 67.3%.

[0029] Content determination: Puerarin was determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020).

[0030] 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-water (25:75) was used as the mobile phase; the detection wavelength was 250 nm. The theoretical plate number, calculated based on the puerarin peak, should not be less than 4000.

[0031] Preparation of reference solution: Take an appropriate amount of puerarin reference standard, accurately weigh it, and add 30% ethanol to prepare a solution containing 80 μg per ml.

[0032] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 0.5g, weigh it accurately, place it in a stoppered conical flask, add 50ml of 30% ethanol accurately, weigh it, heat under reflux for 30 minutes, cool it, weigh it again, make up the weight loss with 30% ethanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0033] Assay: Accurately pipette 5-10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0034]

[0035] This product contains 25mg / g to 37mg / g of puerarin per 1g;

[0036] The transfer rate of puerarin content in the decoction of medicinal materials ranges from 42% to 78%.

[0037] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0038] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase (Shiseido: CAPCELL PAK C18 (5μm, 25×0.46cm), Yuexu: Ultimate XB-C18 (5μm, 25×0.46cm), Shimadzu: Wondasil C18.superb (5μm, 25×0.46cm)); octadecylsilane-bonded silica gel was used as the stationary phase (column length 250mm, inner diameter 4.6mm, particle size 5μm); acetonitrile was used as mobile phase A, and 0.1% formic acid solution was used as mobile phase B, with gradient elution performed according to the specifications in Table 1; the detection wavelength was 250nm. The theoretical plate number, calculated based on the daidzein peak, should not be less than 3000.

[0039] Table 1 shows the gradient elution table for chromatographic conditions and system suitability testing:

[0040]

[0041] Preparation of reference solution: Take about 0.5g of kudzu root reference material, place it in a stoppered conical flask, add 50ml of 30% ethanol, weigh it, heat under reflux for 30 minutes, cool, weigh it again, replenish the lost weight with 30% ethanol, shake well, filter, and take the filtrate as the reference solution. Separately, take an appropriate amount of daidzein reference standard, accurately weigh it, add 30% ethanol to prepare a solution containing 70μg per ml, and use it as the reference solution.

[0042] Preparation of test solution: Same as (content determination).

[0043] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0044] like Figure 2 As shown (peak 1: puerarin; peak 3(S): daidzein; peak 5: daidzein; each milligram of the test sample is approximately equivalent to 0.041 grams of medicinal slices), the characteristic chromatogram of the test sample should show chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the reference medicinal material, and peak 3(S) should be consistent with the retention time of the reference peak; in the chromatogram of the test sample, the retention times of characteristic peaks 1-2 should be within ±4% of the corresponding peaks in the reference medicinal material, and the retention times of characteristic peaks 4-5 should be within ±7% of the corresponding peaks in the reference medicinal material.

[0045] Raw material selection: 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. Kudzu root is a cultivated product, distributed throughout China. It is mainly produced in Guangxi, Guangdong, Sichuan, Shandong, and other regions of my country, where its cultivation has reached a large scale and is also the main source of kudzu root products. Based on a comprehensive evaluation of raw material quality, sustainable resource supply, and other factors, the preliminary producing areas for raw materials used in large-scale production were determined to be Guangxi, Yulin (Guangxi), Zhaoqing (Guangdong), Pingba Town (Hunan), Pingnan (Guigang), and Tengxian (Wuzhou). Ultimately, 18 qualified batches were selected for the study of standard decoctions.

[0046] Research and standard setting on extract yield and transfer rate;

[0047] Table 2 shows the transfer rate comparison of 18 batches of standard kudzu root decoction:

[0048]

[0049]

[0050]

[0051] Based on the content of 18 batches of kudzu root slices, the content of standard decoction, and the yield of extract, the average transfer rate of the standard decoction was calculated to be 60.1%.

[0052] The range calculated using the mean plus or minus 3 times the SD is 56% to 65%.

[0053] The range calculated using 70% to 130% of the mean is 42% to 78%.

[0054] Based on the actual range, it is 54.3% to 69.7%;

[0055] The upper and lower limits of the standard decoction transfer rate were set at 42% to 78%.

[0056] Content Research and Standardization: Modern research shows that kudzu root contains various components, such as isoflavones (including puerarin), triterpenoids, starch, cellulose, and trace elements. Kudzu root has various pharmacological activities, including effects on the cardiovascular system, lowering blood sugar and lipids, anti-oxidative and alcohol-relieving effects, and anti-tumor effects. Among them, puerarin has the effects of dilating coronary arteries, enhancing myocardial contractility, lowering blood lipids, lowering blood pressure, and preventing and treating dizziness, headache, and neck pain caused by hypertension. The 2020 edition of the Chinese Pharmacopoeia lists puerarin as an indicator component for the quality control of kudzu root. Therefore, this study selected puerarin as the quality control indicator component for the standard kudzu root decoction.

[0057] Based on the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules", and combined with data from 18 batches of standard kudzu root decoction, the content limits and upper and lower limits of the standard decoction were calculated.

[0058] The range calculated using the mean plus or minus 3 times the SD is: 25mg to 37mg;

[0059] The range calculated using 70%–130% of the mean is 22 mg–40 mg.

[0060] Based on the actual range of 28.03mg to 34.54mg, it is recommended to use the mean plus or minus 3 times the SD, i.e., 25mg to 37mg, as the upper and lower limits of the standard decoction content.

[0061] Based on the content of 18 batches of kudzu root slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 46.6%.

[0062] The range calculated using the mean plus or minus 3 times the SD is 56% to 65%.

[0063] The range calculated using 70% to 130% of the mean is 42% to 78%.

[0064] Based on the actual range, it is 54.3% to 69.7%;

[0065] The upper and lower limits of the standard decoction transfer rate were set at 42% to 78%.

[0066] Feature mapping research and standard setting: such as Figure 3 As shown, the chromatogram of the standard kudzu root decoction with experimental batch number BT(FG)211001 was selected as the reference chromatogram to establish a control characteristic chromatogram; based on the data analysis results, five common peaks were finally selected as characteristic peaks, and puerarin was selected as the reference peak. The relative retention times of the characteristic peaks in the chromatogram were calculated.

[0067] According to the national drug standard YBZ-PFKL-2021043 for "Pueraria lobata and Cypress Formula Granules", among the five characteristic peaks in the characteristic spectrum of the standard decoction of Pueraria lobata, peak 1 is puerarin, peak 3 is daidzin, and peak 5 is daidzein.

[0068] Table 3 shows the comparison of retention times for 18 batches of standard kudzu root decoction:

[0069]

[0070]

[0071] Table 4 shows the relative retention times of 18 batches of standard kudzu root decoction:

[0072]

[0073]

[0074] The results showed that the characteristic chromatograms of 18 batches of kudzu root standard decoction samples all showed chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the control medicinal material. The relative retention time RSD of peaks 1-5 was less than 2%, indicating that the elution times of each peak were consistent among the batches.

[0075] Based on the test results of 18 batches of standard kudzu root decoction and the retention time data in the repeatability and precision tests of the methodology (see Tables 5 and 6 below), it can be seen that the retention times of peaks 4 and 5 drifted significantly, basically within ±7%, while the retention times of peaks 1 and 2 were more stable, fluctuating within ±4%. Therefore, it is stipulated that: the characteristic chromatogram of the test sample should show chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the reference medicinal material, and peak 3 (S) should be consistent with the retention time of the reference peak; in the chromatogram of the test sample, the retention times of characteristic peaks 1-2 should be within ±4% of the corresponding peaks in the reference medicinal material, and the retention times of characteristic peaks 4-5 should be within ±7% of the corresponding peaks in the reference medicinal material.

[0076] Table 5 is a comparison table of retention times for repeatability:

[0077]

[0078] Table 6 is a retention time comparison table for precision:

[0079]

[0080] Extractives study and standard setting: Extractives were determined using ethanol as the solvent, following the hot extraction method under the "Determination of Alcohol-Soluble Extractives" section of the Chinese Pharmacopoeia 2020, Part IV, General Chapter 2201. The test results for 18 batches of kudzu root standard decoction are shown in Table 7.

[0081] Table 7 is a comparison table of the results of extract determination of 18 batches of kudzu root:

[0082]

[0083]

[0084] The average value of the alcohol-soluble extract from 18 batches of standard decoctions was 51.8%.

[0085] The range calculated using the mean plus or minus 3 times the SD is 50.2% to 53.4%.

[0086] The range calculated using 70% to 130% of the mean is 36.3% to 67.3%.

[0087] Based on the actual range of 51.1% to 52.7%;

[0088] The upper and lower limits of the standard decoction extract were set at 36.3% to 67.3%.

[0089] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0090] 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 any specific implementation. 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. The preparation process of kudzu root standard decoction, characterized in that, Includes the following steps: S1: Place the kudzu root slices in a clay pot, add 9 times the amount of water, and soak for 30 minutes. Bring to a boil over high heat, then simmer over low heat for 30 minutes to obtain one batch of medicinal liquid. Pour out the first batch of medicinal liquid. S2: Add 7 times the amount of water to the clay pot again, bring to a boil over high heat, then simmer over low heat for 20 minutes to obtain a second decoction. Pour out the second decoction. S3: Filter the primary and secondary medicinal solutions obtained from S1 and S2 through a 200-mesh filter, cool the filtrate to room temperature in a cold water bath, and then combine the two filtrates to obtain the kudzu root medicinal solution; S4: The obtained kudzu root extract was concentrated under vacuum at a temperature of 50°C until the weight ratio of the medicinal material to the fluid extract was 1:

1. S5: Based on S4 above, the concentrated liquid is collected and freeze-dried to obtain the standard kudzu root decoction.

2. The preparation process of the standard kudzu root decoction according to claim 1, characterized in that, In step S4, the obtained kudzu root extract is concentrated under vacuum at a temperature of 50°C, with the absolute vacuum maintained at 30–40 mbar.

3. The preparation process of the standard kudzu root decoction according to claim 1, characterized in that, In step S5, concentration and freeze-drying are used to measure the yield and extract index of kudzu root in the decoction.

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