Preparation process of uncaria standard decoction

Abstract

The invention discloses a preparation process of an uncaria standard decoction, and relates to the technical field of traditional Chinese medicine decoction treatment. The preparation method comprises the following steps: firstly, weighing a proper amount of uncaria decoction pieces, putting the uncaria decoction pieces into a marmite, and decocting twice, and comprises the following steps: step 1, adding water and warm water (50-60 DEG C) which are 10-12 times of the uncaria decoction pieces into the marmite for the first time, soaking for 30-60 minutes, boiling with strong fire, and decocting with slow fire for 30-40 minutes; step 2, adding water 8-10 times of the uncaria decoction pieces into the marmite for the second time, boiling with strong fire, and decocting with slow fire for 20-30 minutes; 3, filtering the liquid medicine with 200 meshes, and cooling the filtrate to room temperature in a cold water bath; 4, combining the two filtrates, and carrying out vacuum concentration at the temperature of 60 + / -5 DEG C; and Step 5, concentrating until the weight ratio of the medicinal material to the fluid extract is about 1: 1, collecting a concentrated solution, and freeze-drying to obtain a sample. According to the preparation process of the uncaria standard decoction, an objective quality standard is used for measuring the component transfer condition from decoction pieces to the decoction, 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 decoction processing technology, and in particular relates to the preparation process of standard Uncaria rhynchophylla decoction. Background Technology

[0002] Traditional Chinese medicine decoctions are a widely used form of clinical medication. When developing Kampo (traditional Chinese medicine) preparations, the concept of a "standardized decoction" was proposed as a benchmark to measure the consistency of efficacy between modern industrial preparations and decoctions originating from the traditional agricultural era. Because Kampo medicines utilize ancient prescriptions from classic Chinese medical texts such as *Shanghan Zabing Lun* and *Jinkui Yaolue*, sharing a common origin, the management experience of Kampo medicines is worth learning from.

[0003] Standard decoctions are single-herb Chinese medicinal decoctions prepared using standardized processes, guided by traditional Chinese medicine theory and based on clinical application, while referencing modern extraction methods. They are used for standardized clinical medication, ensuring accuracy and consistency in dosage. The "standard" in standard decoctions primarily encompasses the representativeness of the medicinal materials used, consistency with traditional preparation methods, and rigorous quality control, essentially guaranteeing the stability and uniformity of the prescription's efficacy and quality. Furthermore, standard decoctions do not undergo a drying process, maintaining maximum consistency with traditional decoctions used in clinical practice.

[0004] Standardized decoctions serve as a "bridge" connecting traditional Chinese medicine (TCM) decoction pieces and modern TCM preparations. They provide a reference for controlling the quality of TCM end products, offer tools for standardizing different forms of TCM administration to ensure quality uniformity and efficacy consistency, and provide a reference for evaluating the consistency of product quality among different manufacturers. Therefore, research on the quality standards of standardized TCM decoction pieces will provide a foundation for the formulation of quality standards for all end products derived from decoctions of Chinese medicinal herbs.

[0005] Since hospitals and the market currently lack quality research and standardization for traditional Chinese medicine decoctions to measure their quality and efficacy after preparation, this study takes Uncaria decoction as an example. It specifies key quality indicators such as the main content, characteristic spectrum, and extracts of single Chinese medicinal herbs after decoction preparation, examines the transfer rate, and defines the range of transfer rates of the main medicinal content to assess the effectiveness of the decoction after preparation according to standard methods. Summary of the Invention

[0006] The purpose of this invention is to provide a preparation process for a standard decoction of Uncaria rhynchophylla, thereby solving existing problems.

[0007] To solve the above-mentioned technical problems, the present invention is achieved through the following technical solution:

[0008] This invention relates to a preparation process for a standard decoction of Uncaria rhynchophylla. First, an appropriate amount of Uncaria rhynchophylla slices are weighed, placed in a clay pot, and decocted twice. The process is characterized by the following steps:

[0009] Step 1: Add 10-12 times the amount of water as Uncaria rhynchophylla slices to the clay pot for the first time, soak in warm water (50℃-60℃) for 30-60 minutes, bring to a boil over high heat, then simmer over low heat for 30-40 minutes.

[0010] Step 2: Add 8-10 times the amount of water to the clay pot for the second time, bring to a boil over high heat, then simmer over low heat for 20-30 minutes.

[0011] Step 3: Then filter the above-mentioned medicinal solution through a 200-mesh filter, and cool the filtrate to room temperature in a cold water bath;

[0012] Step 4: Combine the two filtrates and concentrate them under vacuum at a temperature of 60℃±5℃;

[0013] Step 5: Concentrate the extract to a ratio of approximately 1:1 between the medicinal material and the fluid extract. Collect the concentrate and freeze-dry it to obtain the sample.

[0014] Preferably, in Step 4, vacuum concentration is performed under an absolute vacuum of 30–40 mbar.

[0015] The present invention has the following beneficial effects:

[0016] The preparation process of the Uncaria rhynchophylla standard decoction of the present invention has objective quality standards to measure the component transfer after the medicinal slices are converted into decoction, and to ensure the quality and effectiveness of the decoction.

[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 Uncaria rhynchophylla standard decoction of the present invention.

[0020] Figure 2 This is a comparative characteristic chromatogram of the test solution of the Uncaria rhynchophylla standard decoction of the present invention. Detailed Implementation

[0021] 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.

[0022] In the description of this invention, it should be understood that the terms "upper," "middle," "outer," "inner," etc., which indicate orientation or positional relationship, are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the components or elements referred to must have a specific orientation, or be constructed and operated in a specific orientation, and therefore should not be construed as limiting this invention.

[0023] In the description of this invention, it should be noted that, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0024] Example 1:

[0025] Please see Figure 1 As shown, this invention relates to a preparation process for a standard decoction of Uncaria rhynchophylla. First, an appropriate amount of Uncaria rhynchophylla slices are weighed and placed in a clay pot, then decocted twice. The process is characterized by the following steps:

[0026] Step 1: Add 10-12 times the amount of water as Uncaria rhynchophylla slices to the clay pot for the first time, soak in warm water (50℃-60℃) for 30-60 minutes, bring to a boil over high heat, then simmer over low heat for 30-40 minutes.

[0027] Step 2: Add 8-10 times the amount of water to the clay pot for the second time, bring to a boil over high heat, then simmer over low heat for 20-30 minutes.

[0028] Step 3: Then filter the above-mentioned medicinal solution through a 200-mesh filter, and cool the filtrate to room temperature in a cold water bath;

[0029] Step 4: Combine the two filtrates and concentrate them under vacuum at a temperature of 60℃±5℃;

[0030] Step 5: Concentrate the extract to a ratio of approximately 1:1 between the medicinal material and the fluid extract. Collect the concentrate and freeze-dry it to obtain the sample.

[0031] In Step 4, vacuum concentration is carried out under an absolute vacuum of 30–40 mbar.

[0032] Extract yield: After vacuum concentration of the Uncaria rhynchophylla 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:

[0033]

[0034] Ointment yield range: 6.5%-9.5% (national standard), 6.8%-9.4% (internal control).

[0035] Extractives: Determined by hot extraction method according to 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 should not be less than 23.1% (internal control).

[0036] Content determination

[0037] Determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020 Edition).

[0038] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 100 mm, inner diameter 2.1 mm, particle size 2.6 μm); acetonitrile-0.015 mol / L dipotassium hydrogen phosphate solution (adjusted to pH 7.5-7.6 with 2% phosphoric acid) (35:65) was used as the mobile phase, with a flow rate of 0.3 mL / min; column temperature was 25℃; detection wavelength was 246 nm. The theoretical plate number, calculated based on the isorhamnetin peak, should be no less than 5000.

[0039] Preparation of reference solution: Take an appropriate amount of isorhodin reference standard, accurately weigh it, and add methanol to prepare a solution containing 15 μg per ml.

[0040] 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 25ml of methanol accurately, stopper tightly, weigh it, heat under reflux for 30 minutes, cool it, weigh it again, make up the weight loss with methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0041] Assay: Accurately pipette 1 μl of the reference solution and the test solution into the liquid chromatograph and determine.

[0042] Using isorhynchine reference standard as a reference and its corresponding peak as the S peak, calculate the relative retention times of isodehydrorhynchine, dehydrorhynchine, and rhynchine. The relative retention times should be within ±5% of the specified value (if the relative retention time deviates by more than 5%, confirmation with the corresponding substituted reference standard should be used). The relative retention times and relative correction factors are shown in the table below:

[0043] Table 1 shows the relative retention time and relative correction factor comparison table for rhynchophylline:

[0044]

[0045]

[0046] Using the peak area of ​​isorhodin as a control, the contents of dehydrorhodin, isorhodin, rhodin, and isorhodin were calculated by multiplying by a correction factor.

[0047]

[0048] The total amount of dehydrorheumatine, isodehydrorheumatine, rheumatine, and isorheumatine per 1g of this product should be 1.38mg-2.57mg (internal control). The chlorogenic acid content transfer rate of the medicinal materials to the decoction ranges from 12.5% ​​to 25.5% (internal control).

[0049] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0050] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the packing material (column length 150 mm, inner diameter 2.1 mm, particle size 1.6 μm); acetonitrile was used as mobile phase A, and 0.3% phosphoric acid solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 0.3 mL / min; the column temperature was 40 °C; and the detection wavelength was 245 nm. The theoretical plate number, calculated based on the Uncaria rhynchophylline peak, should not be less than 3000.

[0051] Table 2 shows the gradient elution table:

[0052]

[0053]

[0054] Preparation of reference solutions: Take 0.5g of Uncaria rhynchophylla reference material, add 20ml of 70% methanol, sonicate (250W power, 40kHz frequency) for 20 minutes, cool, shake well, filter, and take the filtrate as the reference solution. Separately, take appropriate amounts of catechin reference standard, chlorogenic acid reference standard, dehydrorhynchophylline reference standard, and rhynchophylline reference standard, accurately weigh them, and add methanol to prepare solutions containing 40μg of catechin, 50μg of chlorogenic acid, 40μg of dehydrorhynchophylline, and 0.1mg of rhynchophylline per ml, as reference solutions.

[0055] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 0.1 g, weigh it accurately, place it in a stoppered conical flask, accurately add 25 ml of 70% methanol, stopper tightly, weigh it, sonicate (power 250W, frequency 40kHz) for 30 minutes, cool it, weigh it again, make up the lost weight with 70% methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0056] Determination method: Accurately pipette 1 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0057] The chromatogram of the test sample should show 10 characteristic peaks, and the retention times should correspond to the 10 characteristic peaks in the chromatogram of the reference medicinal material. Specifically, the retention times of peaks 2, 3, 8, and 9 should correspond to the retention times of the peaks in the chromatograms of the catechin, chlorogenic acid, dehydrorheumatoidin, and rheumatoidin reference standards. The peak corresponding to the chlorogenic acid (peak 3) reference standard is peak S1. Calculate the relative retention times of peaks 1 and 4 with peak S1; these relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.65 (peak 1) and 1.13 (peak 4). The peak corresponding to the rheumatoidin (peak 9) reference standard is peak S2. Calculate the relative retention times of peaks 5–7 and peak 10 with peak S2; these relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.66 (peak 5), 0.70 (peak 6), 0.81 (peak 7), and 1.22 (peak 10). Calculate the relative peak areas of peaks 5, 6, 8, 10 and S2. The relative peak area of ​​peak 5 shall not be less than 0.45, the relative peak area of ​​peak 6 shall not be less than 0.80, the relative peak area of ​​peak 8 shall not be less than 0.25, and the relative peak area of ​​peak 10 shall not be less than 0.60.

[0058] Peak 2: Catechin; Peak 3 (S1): Chlorogenic acid; Peak 4: Cryptochlorogenic acid; Peak 7: Isochlorogenic acid B; Peak 8: Dehydrorhegine; Peak 9 (S2): Rhynchophylline; Peak 10: Rhynchophylline.

[0059] Example 2:

[0060] 1. Raw material selection

[0061] 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. Uncaria rhynchophylla (Gou Teng) is a cultivated product, distributed throughout my country. It is mainly produced in Sichuan, Jiangxi, and Guizhou provinces, where its cultivation has reached a large scale and is the main source of commercial Uncaria rhynchophylla. Based on a comprehensive evaluation of raw material quality, sustainable resource supply, and other factors, the preliminary production areas for raw materials used in large-scale production were determined to be Dazhu County, Dazhou City, Sichuan Province; Jiangyou City, Mianyang City, Sichuan Province; Zhangshu City, Yichun City, Jiangxi Province; and Jianhe County, Qiandongnan Miao and Dong Autonomous Prefecture, Guizhou Province. Ultimately, 12 qualified batches were selected for research on standard decoctions.

[0062] 2. Research and standard setting on extract yield and transfer rate

[0063] Fifteen batches of Uncaria rhynchophylla slices were taken and prepared into standard decoction samples according to the standard decoction preparation method. The yield of extract and the content of Uncaria rhynchophylla alkaloids in each standard decoction were determined, and the transfer rate was calculated according to the following formula. The results are shown in Table 2-9. (Content A includes: isorhodin, isorhodin, deorhodin, and rhodin.)

[0064]

[0065] Table 2-9 Transfer rate of 12 batches of Radix Paeoniae Alba Standard Decoction

[0066]

[0067] Based on the total content of 12 batches of Uncaria rhynchophylla slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 19.0%.

[0068] The range calculated using the mean plus or minus 3 times the SD is 12.5% ​​to 25.5%.

[0069] The range calculated using 70% to 130% of the mean is 13.3% to 24.8%.

[0070] Based on the actual range, it is 16.6% to 24.8%;

[0071] It is recommended to take 12.5% ​​to 25.5% as the upper and lower limits of the standard decoction transfer rate.

[0072] Standard method for measuring the transfer rate of decoctions:

[0073] σ: Standard deviation of the transfer rate of standard decoction

[0074] The Bonett method is effective for any continuous distribution.

[0075] The chi-square method is only effective for normal distributions.

[0076] Descriptive statistics

[0077]

[0078] Statistical analysis of the transfer rate (total alkaloids from Uncaria rhynchophylla): There were no significant differences in the (transfer rate - total amount) among the various batches of standard decoctions. Normal distribution analysis showed a p-value of 0.896 (p > 0.05), meeting the conditions for normal distribution. Analysis of variance under normal distribution showed a variance value of 0.00171 (variance < 0.05). Therefore, there were no significant differences in the (transfer rate - total amount) among the various batches of standard decoctions corresponding to different batches of medicinal materials. This proves that the (transfer rate - total amount) of the standard decoctions within the scope of this study is controllable, and that there are no significant differences in the (transfer rate - total amount) after Uncaria rhynchophylla from different producing areas is processed into standard decoctions. The total amount range is highly controllable and acceptable, and can be used as the (transfer rate) item for standard decoctions in standardization.

[0079] 3. Content research and standard setting

[0080] Modern research shows that Uncaria rhynchophylla contains a variety of chemical components, mainly alkaloids (alkaloids are abundant in Uncaria rhynchophylla and are also important active ingredients for its pharmacological effects. The main alkaloids in Uncaria rhynchophylla are indole alkaloids and oxyindole alkaloids, of which rhynchophylline accounts for 28.9% of the total alkaloids and isorhynchophylline accounts for 14.7%, accounting for more than 40% of the total alkaloids), flavonoids (9 flavonoid compounds including kaempferol, quercetin, isoquercetin, and (1)-epicatechin rutin), terpenes, esters, and other active ingredients. Uncaria rhynchophylla (Gou Teng) possesses various pharmacological activities, including hypotensive effects (Uncaria alkaloids can directly lower blood pressure by dilating blood vessels, reducing cardiac output, and decreasing the influx of exogenous calcium ions into tissues, and indirectly by blocking nerve conduction and reducing neurotransmitter secretion), effects on the central nervous system (sedative, anticonvulsant, and antiepileptic effects; Uncaria alkaloids also regulate the central dopamine (DA) system), protective effects against cerebral ischemia-reperfusion injury, anticancer effects, anti-inflammatory effects, and analgesic effects. The 2020 edition of the Chinese Pharmacopoeia does not specify quality control indicators. Referring to the "Standard for Uncaria rhynchophylla (Gou Teng) Formula Granules" YBZ-PFKL-2021052, which specifies the content of isorhodin, isorhodin, dehydrorhodin, and the total amount of rhodin, this study selected isorhodin, isorhodin, dehydrorhodin, and the total amount of rhodin as the quality control indicators for standard Uncaria rhynchophylla (Gou Teng) decoctions.

[0081] According to the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules", and based on the data of 12 batches of Uncaria rhynchophylla (Gou Teng) standard decoction, the content limit and upper and lower limits of the standard decoction were calculated, and the average value was 1.98 mg.

[0082] The range calculated using the mean plus or minus 3 times the SD is: 1.61mg-2.34mg;

[0083] The range calculated using 70%–130% of the mean is: 1.38 mg–2.57 mg ；

[0084] Based on the actual range of 1.78mg to 2.16mg, it is recommended to use 70%-130% of the average value.

[0085] The upper and lower limits of the standard decoction content are 1.38mg-2.57mg.

[0086] Statistical analysis of the content (Uncaria alkaloids): There were no significant differences in the content determination results of Uncaria alkaloids among the various batches of standard decoctions. Normal distribution analysis showed a p-value of 0.495 (P > 0.05), meeting the conditions for normal distribution. Analysis of variance under normal distribution showed a variance value of 0.0147 (Variance < 0.05). Therefore, there were no significant differences in the content determination of Uncaria alkaloids among the various batches of standard decoctions corresponding to different batches of medicinal materials. This proves that the content determination of Uncaria alkaloids in the standard decoctions within the scope of this study is controllable, and that there are significant differences in the content determination of Uncaria alkaloids after processing Uncaria from different producing areas into standard decoctions. Further analysis of the transfer rate of the standard decoctions is needed to determine the reliability of the standard decoction process.

[0087] 4. Characteristic spectrum research and standard setting

[0088] The chromatogram of the Uncaria rhynchophylla standard decoction with batch number BT(TR)01 was selected as the reference chromatogram. Based on the data analysis results, 10 common peaks were ultimately selected as characteristic peaks. The generated standard chromatogram is shown below. Comparison of the characteristic chromatogram with the reference standard chromatogram revealed that among the 10 characteristic peaks of the Uncaria rhynchophylla standard decoction, those specified in the "Uncaria rhynchophylla Formula Granules" were...

[0089] YBZ-PFKL-2021052 indicates that (peak 2: catechin; peak 3 (S1): chlorogenic acid; peak 4: cryptochlorogenic acid; peak 7: isochlorogenic acid B; peak 8: dehydrorhegine; peak 9 (S2): rhegine; peak 10: picroside).

[0090] Retention time of 12 batches of standard decoction of Uncaria rhynchophylla (peak 1 to peak 5)

[0091]

[0092] Retention time of 12 batches of standard decoction of Uncaria rhynchophylla (peak 6-peak 10)

[0093]

[0094] Relative retention time of 12 batches of standard decoction of Uncaria rhynchophylla (Gou Teng)

[0095]

[0096]

[0097] Peak area of ​​12 batches of Uncaria (Gou Teng) standard decoction

[0098]

[0099] Relative peak area of ​​12 batches of Uncaria (Gou Teng) standard decoction

[0100]

[0101] The results showed that the characteristic chromatograms of all 12 batches of Uncaria rhynchophylla (Gou Teng) standard decoction samples exhibited chromatographic peaks corresponding to the retention times of the 10 main characteristic peaks in the reference herb. Seven peaks (peak 2: catechin; peak 3 (S1): chlorogenic acid; peak 4: cryptochlorogenic acid; peak 7: isochlorogenic acid B; peak 8: dehydrorhynchophylline; peak 9 (S2): rhynchophylline; peak 10: picroside) should correspond to the retention times of the corresponding reference peaks. The relative retention times RSD of peaks 1 and 4 with peak 3 (S1), and peaks 5, 6, 7, and 10 with peak 9 (S2) were less than 2%. The relative peak areas of peaks 5, 6, 8, and 10 with peak S2 were calculated; the relative peak area of ​​peak 5 should not be less than 0.45, the relative peak area of ​​peak 6 should not be less than 0.80, the relative peak area of ​​peak 8 should not be less than 0.25, and the relative peak area of ​​peak 10 should not be less than 0.60.

[0102] The national standard YBZ-PFKL-2021052, "Uncaria rhynchophylla Formula Granules," only specifies the retention times of reference standards for peaks 2, 3, 8, and 9; the relative retention times of peaks 1, 4, and 3 (S1); peaks 5, 6, 7, and 10 and peak 9 (S2); and the calculation of the relative peak areas of peaks 5, 6, 8, 10, and S2. Based on the determination results of 12 batches of Uncaria rhynchophylla (Gou Teng) standard decoction and the retention time data in the repeatability and precision tests of the methodology, it can be seen that the relative peak drift of peaks 1, 4 and 3 (S1), 5, 6, 7, 10 and 9 (S2) is very small, basically within ±5%. The relative peak areas of peaks 5, 6, 8, 10 and S2 were calculated; the relative peak area of ​​peak 5 should not be less than 0.45, peak 6 should not be less than 0.80, peak 8 should not be less than 0.25, and peak 10 should not be less than 0.60. Therefore, it is stipulated that the chromatogram of the test sample should show 10 characteristic peaks, and the retention times should correspond to the 10 characteristic peaks in the chromatogram of the reference medicinal material. Among them, the retention times of peaks 2, 3, 8, and 9 should correspond to the retention times of the reference peaks in the chromatograms of catechin, chlorogenic acid, dehydrouncitonine, and uncitonine. The peak corresponding to the chlorogenic acid (peak 3) reference is peak S1. Calculate the relative retention times of peaks 1 and 4 with peak S1. These relative retention times should be within ±10% of the specified values, which are: 0.65 (peak 1) and 1.13 (peak 4). The peak corresponding to the rhizoside (peak 9) reference is peak S2. Calculate the relative retention times of peaks 5-7 and peak 10 with peak S2. These relative retention times should be within ±10% of the specified values, which are: 0.66 (peak 5), 0.70 (peak 6), 0.81 (peak 7), and 1.22 (peak 10). Calculate the relative peak areas of peaks 5, 6, 8, and 10 with peak S2. The relative peak area of ​​peak 5 should not be less than 0.45, peak 6 should not be less than 0.80, peak 8 should not be less than 0.25, and peak 10 should not be less than 0.60.

[0103] 5. Leachate research and standard setting

[0104] According to the extractives section of the "Uncaria rhynchophylla (Gnaphalium affine) Formula Granules" standard YBZ-PFKL-2021052, the finished granule product is required to include an extractives determination item. Refer to the method for determining the extractives of the finished granule product.

[0105] Results of extract analysis of 12 batches of Uncaria rhynchophylla (Gou Teng)

[0106]

[0107] The average value of the alcohol-soluble extract from 12 batches of standard decoction was 26.8%.

[0108] The range calculated using the mean plus or minus 3 times the SD is 23.1% to 30.5%.

[0109] The range calculated using 70% to 130% of the mean is 18.8% to 34.9%.

[0110] Based on the actual range of 25.2% to 29.1%;

[0111] It is recommended to take 23.1% to 30.5% as the upper and lower limits of the standard decoction extract.

[0112] Method for extracting standard broth:

[0113] σ: Standard deviation of standard decoction extract

[0114] The Bonett method is effective for any continuous distribution.

[0115] The chi-square method is only effective for normal distributions.

[0116] Descriptive statistics

[0117]

[0118] Statistical analysis of extracts: There were no significant differences in the extracts measured among the various batches of standard decoctions. Normal distribution analysis showed a p-value of 0.562 (P > 0.05), meeting the conditions for normal distribution. Analysis of variance under normal distribution showed a variance value of 0.000154 (Variance < 0.05). Therefore, there were no significant differences in the extracts among the various batches of standard decoctions corresponding to different batches of medicinal materials. This demonstrates that the extracts in the standard decoctions within the scope of this study are controllable, and that there are no significant differences in the extracts of Uncaria rhynchophylla from different producing areas after being processed into standard decoctions. The established extract range is highly controllable and acceptable, and can be used as an extract item for standard decoctions in standard formulation.

[0119] In the description of this specification, references to terms such as "an embodiment," "example," and "specific example" 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, 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.

[0120] 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. The preparation process of the standard decoction of Uncaria rhynchophylla, firstly, weigh the Uncaria rhynchophylla slices, place them in a clay pot, and decoct twice, characterized in that... Includes the following steps: Step 1: Add 10-12 times the amount of water to the clay pot for the first time, soak in warm water for 30-60 minutes, bring to a boil over high heat, then simmer over low heat for 30-40 minutes. Step 2: Add 8-10 times the amount of water to the clay pot for the second time, bring to a boil over high heat, then simmer over low heat for 20-30 minutes. Step 3: Then filter the above-mentioned medicinal solution through a 200-mesh filter, and cool the filtrate to room temperature in a cold water bath; Step 4: Combine the two filtrates and concentrate them under vacuum at a temperature of 60℃±5℃; Step 5: Concentrate the extract to a ratio of approximately 1:1 between the medicinal material and the fluid extract. Collect the concentrate and freeze-dry it to obtain the sample.

2. The preparation process of the Uncaria standard decoction according to claim 1, characterized in that, In Step 4, vacuum concentration is carried out under an absolute vacuum of 30–40 mbar.

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