Preparation process of golden cypress standard decoction

Abstract

The invention discloses a preparation process of a golden cypress standard 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 cortex phellodendri decoction pieces, putting the cortex phellodendri decoction pieces into a marmite, decocting twice, adding 10 times of water for soaking for 30 minutes for the first time, boiling with strong fire, and then decocting with slow fire for 30 minutes; adding 8 times of water for the second time, boiling with strong fire, then decocting with slow fire for 20 minutes, filtering the liquid medicine with 200 meshes, rapidly cooling the filtrate to room temperature in a water bath, mixing the two filtrates, carrying out vacuum concentration at the temperature of 50 DEG C until the weight ratio of the medicinal material to the fluid extract is about 1: 1, collecting the concentrated solution, and freeze-drying to obtain a sample. According to the invention, the golden cypress decoction is taken as a standard, key quality indexes such as the main content, the characteristic spectrum and the extract of the traditional Chinese medicine single variety after the decoction is prepared are stipulated, the transfer rate is investigated, the transfer rate range of the main content of the golden cypress medicinal material is stipulated, and the effectiveness of the golden cypress decoction is ensured after the golden cypress decoction is prepared according to the standard.

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 standard Phellodendron bark decoction. Background Technology

[0002] Phellodendron bark is the dried bark of the Phellodendron amurense tree (Rutaceae family). It is in the form of plates or shallow grooves, varying in length and width, and 1-6 mm thick. The outer surface of Phellodendron bark is yellowish-brown or yellowish-brown, flat or with longitudinal grooves, and sometimes lenticel scars and remnants of grayish-brown rough bark are visible. The inner surface is dark yellow or light brown, with fine longitudinal ridges. It has the effects of clearing heat and drying dampness, purging fire and eliminating steaming, and detoxifying and healing sores. Traditional Chinese medicine decoction is a traditional and widely used form of medication in clinical practice. Standard decoction is a single-herb Chinese medicine decoction prepared by standardized processes based on traditional Chinese medicine theory and clinical application, with reference to modern extraction methods. It is 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 Phellodendron amurense decoction, so it is impossible to measure the quality and efficacy of the decoction after decoction, nor can it provide a basis for the formulation of quality standards for the final product of Phellodendron amurense decoction. Therefore, we propose a preparation process for a standard Phellodendron amurense decoction. Summary of the Invention

[0004] The purpose of this invention is to provide a preparation process for a standard decoction of Phellodendron bark, so as 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 decoction of Phellodendron bark, which includes two decoction processes. The preparation process includes the following steps:

[0007] S1: Place the Huangbai (Phellodendron bark) slices in a clay pot, add 10 times the amount of water, and soak for 30 minutes. Bring to a boil over high heat, then reduce to low heat and simmer for 30 minutes to obtain a first decoction. Pour out the first decoction.

[0008] S2: Add 8 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 first and second medicinal solutions obtained in the above steps 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 Phellodendron bark medicinal solution;

[0010] S4: The obtained Phellodendron bark 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 decoction of Phellodendron bark.

[0012] Among them, the maximum vacuum level that can be achieved by the oil-free variable frequency diaphragm vacuum pump is used as the vacuum environment, that is, the filtrate is concentrated under vacuum conditions of 50°C and absolute vacuum of 30-40 mbar.

[0013] Concentration and freeze-drying are used to measure the yield and extract indicators of Phellodendron bark in decoctions.

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

[0015] The preparation process of the standard decoction of Phellodendron bark in this invention uses objective quality standards to measure the component transfer of Phellodendron bark slices into decoction, and ensures the quality and effectiveness of the Phellodendron bark decoction.

[0016] The present invention discloses the preparation process of the standard decoction of Phellodendron bark and specifies the quality standards of the decoction, including content determination, characteristic chromatogram, extract index, and content transfer rate of the decoction.

[0017] The "standard" in the Huangbai 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 Huangbai prescriptions. In addition, the Huangbai standard decoction has not undergone a drying process, thus maintaining the consistency with the traditional decoction in clinical practice to the greatest extent.

[0018] 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

[0019] 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.

[0020] Figure 1 This is a flowchart illustrating the preparation process of the standard decoction of Phellodendron bark according to the present invention;

[0021] Figure 2 This is a characteristic peak comparison spectrum of the present invention;

[0022] Figure 3 This is a comparative characteristic spectrum of the standard decoction of Phellodendron bark in this invention. Detailed Implementation

[0023] 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.

[0024] Please refer to Figure 1 As shown, this invention relates to a preparation process for a standard decoction of Phellodendron bark, the method of which includes:

[0025] Weigh an appropriate amount of Phellodendron bark slices, place them in a clay pot, and decoct twice. For the first decoction, add 10 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 8 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 sieve, 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.

[0026] Extract yield: After vacuum concentration of the Phellodendron amurense 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:

[0027]

[0028] Ointment yield range: 13%–19%.

[0029] Extractives: Determined by hot extraction method according to the method for determination of alcohol-soluble extractives (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201) using ethanol as solvent. The extractives range of the standard decoction is 34.4% to 63.3%.

[0030] Content determination: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 edition, Part IV, General Chapter 0512).

[0031] Berberine hydrochloride:

[0032] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-0.1% phosphoric acid (11:89) was used as the mobile phase; the detection wavelength was 326 nm. The theoretical plate number, calculated based on the berberine hydrochloride peak, should not be less than 5000.

[0033] Preparation of reference solution: Weigh an appropriate amount of berberine hydrochloride reference standard accurately, place it in a brown volumetric flask, and add methanol to prepare a solution containing 40 μg per ml.

[0034] Preparation of the test solution: Grind the sample into a fine powder, weigh approximately 0.2 g, place it in a stoppered conical flask, accurately add 50 ml of 50% methanol, seal tightly, weigh, sonicate (power 250 W, frequency 50 kHz) for 30 minutes, cool, weigh again, replenish the lost weight with 50% methanol, shake well, filter, and take the filtrate, place it in a brown bottle, and the test solution is obtained.

[0035] Determination method: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0036]

[0037] This product contains 98mg / g to 182mg / g of berberine hydrochloride per 1g.

[0038] The transfer rate of berberine hydrochloride content in the decoction of medicinal materials ranges from 23% to 43%.

[0039] Phellodendron hydrochloride:

[0040] Chromatographic conditions: C18 Superb column, 5μm, 4.6×250mm; mobile phase: acetonitrile-0.1% phosphoric acid solution (0.2g sodium dodecyl sulfonate per 100ml) (36:64); flow rate: 1.0ml / min; column temperature: 30℃; detection wavelength: 284nm. The theoretical plate number, calculated based on the berberine hydrochloride peak, should be no less than 6000.

[0041] Preparation of reference solution: Take an appropriate amount of berberine hydrochloride reference standard, accurately weigh it, and add methanol to prepare a solution containing 0.4 mg per ml.

[0042] 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 50% methanol, weigh it, sonicate it (power 250W, frequency 40kHz) for 20 minutes, cool it, weigh it again, add 50% methanol to make up the weight loss, shake it well, filter it, and take the filtrate to obtain the test solution.

[0043] Assay: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph and determine.

[0044] This product contains 11.3 mg / g to 24.7 mg / g of berberine hydrochloride per 1g.

[0045] The transfer rate of berberine hydrochloride content in the decoction of medicinal materials ranges from 44% to 63%.

[0046] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0047] 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)); acetonitrile was used as mobile phase A, and 0.1% phosphoric acid solution was used as mobile phase B. Elution was performed according to the gradient table in Table 1, with a wavelength of 225nm. The theoretical plate number, calculated based on the berberine hydrochloride peak, should not be less than 5000.

[0048] Table 1 shows the gradient elution table for chromatographic conditions and system suitability testing:

[0049]

[0050] Preparation of reference solution: Take 1g of Phellodendron bark reference material, place it in an Erlenmeyer flask, add 25ml of water, weigh it, heat under reflux for 30min, remove it, let it cool, weigh it again, replenish the lost weight with water, shake well, filter, and you will get the Phellodendron bark reference solution.

[0051] Reference solution: Take the reference solution from the [Assay] section.

[0052] Preparation of the test solution: Take the test solution from the [Assay] section to obtain the solution.

[0053] Determination method: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph, measure and record the chromatogram to obtain the result.

[0054] like Figure 2 As shown (each milligram of the test sample is approximately equivalent to 0.035 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 4 (S) should be consistent with the retention time of the reference peak; in the chromatogram of the test sample, the retention time of characteristic peak 1 should be within ±8% of the corresponding peak in the reference medicinal material, and the retention times of characteristic peaks 2 and 3 should be within ±4% of the corresponding peaks in the reference medicinal material. The repeatability and precision tests of the relative peak areas of peaks 1, 2, 3, and 5 show significant differences. Based on the detection limits of 18 batches of standard decoction, the specified ranges are: peak 1 not less than 0.10, peak 2 0.24–0.42, and peak 3 not less than 0.10.

[0055] 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. Phellodendron bark is a cultivated product, distributed throughout my country. It is mainly produced in Guizhou, Hubei, and Sichuan provinces, where its cultivation has reached a large scale and is the main source of commercial Phellodendron bark. Based on a comprehensive evaluation of raw material quality, sustainable resource supply, and other factors, the preliminary producing areas for large-scale production were determined to be Bingchuan County in Shifang City, Sichuan Province; Kaili in Guizhou Province; Jianshi County in Enshi Prefecture, Hubei Province; and Wufeng County in Yichang City, Hubei Province. Ultimately, 18 qualified batches were selected for the study of standard decoctions.

[0056] Research and standard setting on extract yield and transfer rate:

[0057] Table 2 shows the transfer rate (berberine hydrochloride) of 18 batches of standard decoction of Phellodendron bark:

[0058]

[0059]

[0060] Based on the content of 18 batches of Phellodendron bark, the content of standard decoction, and the yield of extract, the average value of the transfer rate of standard decoction content (berberine hydrochloride) was calculated to be 33.4%. The upper and lower limits calculated by the two methods are slightly different. The range calculated using 70% to 130% of the mean (23% to 43%) is slightly narrower than the range calculated using the mean ± 3 times SD (21% to 46%). The actual range is 26% to 40%. It is recommended to take 23% to 43% as the upper and lower limits of the transfer rate of standard decoction.

[0061] Table 3 shows the transfer rate (berberine hydrochloride) of 18 batches of standard Phellodendron amurense decoction:

[0062]

[0063]

[0064] Based on the content of 18 batches of Phellodendron bark, the content of standard decoction, and the yield of extract, the average value of the transfer rate of standard decoction content (Phellodendron hydrochloride) was calculated to be 53%. The upper and lower limits calculated by the two methods are slightly different. The range calculated using 70% to 130% of the mean (44% to 63%) is slightly narrower than the range calculated using the mean ± 3 times SD (37% to 69%), and the actual range is 49% to 59%. It is recommended to take 44% to 63% as the upper and lower limits of the transfer rate of standard decoction.

[0065] Content Research and Standard Setting: Modern research shows that Phellodendron bark contains various components, such as monoterpenes and their glycosides, triterpenes, mixed-source terpenes, phenylethanol glycosides, volatile oils, flavonoids, tannins, steroids, and organic acids. Phellodendron bark has pharmacological effects such as anti-inflammatory, hepatoprotective, analgesic, and blood-nourishing properties. Berberine hydrochloride is the effective component of Phellodendron bark for hepatoprotection, anti-inflammatory, and analgesic effects. The 2020 edition of the Chinese Pharmacopoeia lists berberine hydrochloride as an indicator component for the quality control of Phellodendron bark; therefore, this study selected berberine hydrochloride as the quality control indicator component for the standard decoction of Phellodendron bark.

[0066] According to the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules," and based on data from 18 batches of Phellodendron amurense standard decoctions, the average berberine hydrochloride content in these 18 batches was calculated to be 140 mg / g. The 70%–130% range of the average is 98 mg / g–182 mg / g, and the SD (mean ± 3) is 32 mg / g–248 mg / g, with an actual range of 62 mg / g–168 mg / g. Because the upper and lower limits calculated using the SD (mean ± 3) are nearly eight times different, this method is not recommended. Considering both the 70%–130% average and the actual range, a range of 98 mg / g–182 mg / g is suggested as the upper and lower limits for the standard decoction content.

[0067] The calculated average content of berberine hydrochloride in 18 batches of standard decoctions was 19.0 mg / g. The 70%–130% range of the average was 13.3 mg / g–24.7 mg / g, and the range was 7.0 mg / g–31.0 mg / g (mean ± 3 times SD). The actual range was 11.2 mg / g–23.9 mg / g. Because the upper and lower limits calculated using the mean ± 3 times SD differ by nearly five times, this method is not recommended. Considering both the 70%–130% average and the actual range, a range of 11.3 mg / g–24.7 mg / g is suggested as the upper and lower limits for the standard decoction content.

[0068] Based on the content of 18 batches of Phellodendron bark, the content of standard decoction, and the yield of extract, the average value of the transfer rate of standard decoction content (berberine hydrochloride) was calculated to be 33.4%. The upper and lower limits calculated by the two methods are slightly different. The range calculated using 70% to 130% of the mean (23% to 43%) is slightly narrower than the range calculated using the mean ± 3 times SD (21% to 46%). The actual range is 26% to 40%. It is recommended to take 23% to 43% as the upper and lower limits of the transfer rate of standard decoction.

[0069] Based on the content of 18 batches of Phellodendron bark, the content of standard decoction, and the yield of extract, the average value of the transfer rate of standard decoction content (Phellodendron hydrochloride) was calculated to be 53%. The upper and lower limits calculated by the two methods are slightly different. The range calculated using 70% to 130% of the mean (44% to 63%) is slightly narrower than the range calculated using the mean ± 3 times SD (37% to 69%), and the actual range is 49% to 59%. It is recommended to take 44% to 63% as the upper and lower limits of the transfer rate of standard decoction.

[0070] Feature mapping research and standard setting

[0071] Using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (Version A)," the chromatogram of the standard decoction of Phellodendron amurense (batch number BT(HB)210801) was selected as the reference chromatogram. The median method was used to generate the control fingerprint chromatogram, with a time window width of 0.50 min, to establish the control characteristic chromatogram. Based on the data analysis results, five common peaks were finally selected as characteristic peaks. The generated standard chromatogram is shown below. Figure 3 Berberine hydrochloride was selected as the reference peak, and the relative retention times of the characteristic peaks in the chromatogram were calculated. Comparison with the chromatogram of the reference standard revealed that peak 4(S) of the five characteristic peaks in the chromatogram of the Huangbai standard decoction was berberine hydrochloride. According to the national drug standard YBZ-PFKL-2021049 for Huangbai formula granules, peak 2: 5-O feruloylquinic acid; peak 4(S): berberine hydrochloride; peak 5: berberine hydrochloride.

[0072] Table 4 shows the relative retention times of 18 batches of standard Phellodendron bark decoction:

[0073]

[0074]

[0075] Table 5 shows the relative peak areas of 18 batches of standard Phellodendron bark decoctions:

[0076]

[0077]

[0078] The results showed that the characteristic chromatograms of 18 batches of Phellodendron bark standard decoction samples all showed chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the reference medicinal materials. The relative retention time RSD of peaks 1-5 was less than 2%, indicating that the elution times of each peak were consistent among different batches.

[0079] The relative peak area RSDs of peaks 1, 2, and 3 are all greater than 10%, indicating that there are significant differences in the relative peak areas of each peak among different batches. Based on the detection limits of 18 batches of standard soup, the specified ranges are: peak 1 not less than 0.10, peak 2 0.24–0.42, and peak 3 not less than 0.10.

[0080] Based on the test results of 18 batches of Phellodendron bark standard decoction and the retention time data in the repeatability and precision tests of the methodology (see Tables 6, 7, 8, and 9 below), it can be seen that the retention time of peak 2 has a large drift, basically within ±8%, while the retention times of peaks 1 and 3 are relatively 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 4 (S) should be consistent with the retention time of the reference peak; in the chromatogram of the test sample, the retention time of characteristic peak 1 should be within ±8% of the corresponding peak in the reference medicinal material, and the retention times of characteristic peaks 1 and 3 should be within ±4% of the corresponding peaks in the reference medicinal material. The repeatability and precision tests of the relative peak areas of peaks 1, 2, 3 and 5 showed significant differences. Based on the detection limits of 18 batches of standard soup, the specified ranges are: peak 1 not less than 0.10, peak 2 0.24 to 0.42, and peak 3 not less than 0.10.

[0081] Table 6 is a comparison table of relative retention times for repeatability:

[0082]

[0083] Table 7 is a comparison table of reproducible relative peak areas:

[0084]

[0085] Table 8 is a retention time comparison table for precision:

[0086]

[0087]

[0088] Table 9 is a comparison table of relative peak areas for precision:

[0089]

[0090] 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 Phellodendron amurense standard decoction are shown in Table 10.

[0091] Table 10 is a comparison table of the results of extract determination of 18 batches of Phellodendron bark:

[0092]

[0093]

[0094] The alcohol-soluble extract of 18 batches of standard decoction was determined, with an average value of 49.1%. The range calculated using the mean plus or minus 3 times the SD was 47.3%–50.9%; the range calculated using the mean of 70%–130% was 34.4%–63.3%; combined with the actual range of 48.0%–50.3%, 34.4%–63.3% was taken as the upper and lower limits of the extract of the standard decoction.

[0095] 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.

[0096] 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.

[0097] 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 standard Phellodendron bark decoction, characterized in that, The preparation process includes two decoctions and includes the following steps: S1: Place the Huangbai (Phellodendron bark) slices in a clay pot, add 10 times the amount of water, and soak for 30 minutes. Bring to a boil over high heat, then reduce to low heat and simmer for 30 minutes to obtain a first decoction. Pour out the first decoction. S2: Add 8 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 first and second medicinal solutions obtained in the above steps 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 Phellodendron bark medicinal solution; S4: The obtained Phellodendron bark 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 decoction of Phellodendron bark.

2. The preparation process of the standard decoction of Phellodendron bark according to claim 1, characterized in that, In step S4, the obtained Phellodendron bark 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 decoction of Phellodendron bark according to claim 1, characterized in that, In S5, concentration and freeze-drying are used to measure the yield and extract index of Phellodendron bark in decoction.

Images