Preparation process of white mulberry root-bark standard decoction

Abstract

The invention discloses a preparation technology of a white mulberry root-bark standard decoction, and relates to the technical field of traditional Chinese medicine decoction preparation, the preparation technology comprises the following steps: weighing white mulberry root-bark decoction pieces, placing in a marmite, adding 10 times of water, soaking for 30 min, boiling with strong fire, and decocting with slow fire for 30 min to obtain a mixed liquid medicine; adding 8 times of water again, boiling with strong fire, and decocting with slow fire for 20 minutes to obtain further mixed liquid medicine; based on the steps, the mixed liquid medicine is filtered by 200 meshes, the filtrate is cooled to room temperature in a cold bath, and the two filtrates are combined; and carrying out vacuum concentration at the temperature of 50 DEG C until the weight ratio of the medicinal material to the fluid extract is 1: 1, collecting the concentrated solution, and freeze-drying to obtain the sample. According to the preparation process of the white mulberry root-bark standard decoction, key quality indexes such as the main content, the characteristic spectrum and the extract of the white mulberry root-bark after the decoction is prepared are stipulated.

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 mulberry bark standard decoction. Background Technology

[0002] Mulberry bark is the dried root bark of the mulberry tree (Morus alba) in the Moraceae family (dicotyledonous plant) after removing the cork. First recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), it is sweet and cold in nature, and enters the lung and spleen meridians. Mulberry bark has the effects of purging lung heat and relieving asthma, promoting diuresis and reducing swelling. Its main functions include: purging lung heat and relieving asthma, promoting diuresis and reducing swelling, tonifying deficiency, promoting blood circulation and removing blood stasis, resolving phlegm and stopping cough, stopping bleeding, promoting body fluid production and quenching thirst, purging and relieving constipation, expelling parasites, aiding digestion, dispelling wind, promoting urination, and relieving pain. Traditional Chinese medicine decoctions are a widely used form of medication in clinical practice. Standard decoctions are single-herb Chinese medicinal decoctions prepared using standardized processes, guided by traditional Chinese medicine theory and based on clinical application, referencing 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 standard regulations for the standard decoction of mulberry bark to measure the quality and efficacy of the decoction after decoction. Therefore, it is impossible to provide a basis for the formulation of quality standards for the final product derived from the decoction of medicinal slices. To this end, we propose a preparation process for the standard decoction of mulberry bark. Summary of the Invention

[0004] The main objective of this invention is to provide a preparation process for a standard decoction of mulberry bark, which will provide a basis for the formulation of quality standards for all final products derived from decoctions of medicinal slices.

[0005] To achieve the above objectives, the technical solution adopted by this invention is as follows: the preparation process of mulberry bark standard decoction includes the following steps:

[0006] S1: Weigh out the mulberry bark slices, place them in a clay pot, 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 to obtain a mixed medicinal liquid.

[0007] S2: Add 8 times the amount of water again, bring to a boil over high heat, then simmer over low heat for 20 minutes to obtain a further mixed medicinal liquid;

[0008] S3: Based on the above steps, filter the mixed solution through a 200-mesh filter, cool the filtrate to room temperature in a cold water bath, and combine the two filtrates;

[0009] S4: Vacuum concentration is carried out at a temperature of 50℃ until the weight ratio of medicinal material to fluid extract is 1:1. The concentrated liquid is then collected and freeze-dried to obtain the sample.

[0010] In the S5 process, vacuum concentration is performed at a temperature of 50°C, with an absolute vacuum of 30-40 mbar.

[0011] In step S5, the extraction rate and extract indicators of mulberry bark in decoction are measured through concentration and freeze-drying operations.

[0012] The present invention has the following beneficial effects:

[0013] The present invention discloses a standard decoction preparation process for mulberry bark, which specifies key quality indicators such as the main content, characteristic spectrum, and extract of mulberry bark after decoction preparation, and examines the transfer rate. By specifying the transfer rate range of the main content of mulberry bark, the effectiveness of mulberry bark decoction is guaranteed after preparation according to the standard method.

[0014] 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

[0015] Figure 1 This is a flowchart illustrating the preparation process of the standard decoction of mulberry bark according to the present invention.

[0016] Figure 2 This is a characteristic peak-to-peak value comparison spectrum diagram of the present invention;

[0017] Figure 3 This is a comparative characteristic chromatogram of the standard decoction of mulberry bark of the present invention. Detailed Implementation

[0018] The technical solutions in this utility model will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0019] like Figure 1 As shown: The preparation process of the standard decoction of mulberry bark is as follows: Weigh an appropriate amount of mulberry 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, and 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, 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 them under vacuum at 50°C 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.

[0020] The mulberry bark aqueous extract was concentrated under vacuum, then freeze-dried to dryness. The dried extract powder was collected, weighed, and the yield was calculated. The formula for the yield is as follows:

[0021]

[0022] Ointment yield range: 13%–21%.

[0023] Its extract was determined using ethanol as the solvent, according to the hot extraction method under the "Determination of Alcohol-Soluble Extracts" (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201). The extract range of the standard decoction was 25.0%–40.3%.

[0024] The content determination procedure includes determining morin A by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020 Edition);

[0025] Chromatographic conditions and system suitability tests were conducted using octadecylsilane-bonded silica gel as the packing material (column length 250 mm, inner diameter 4.6 mm, particle size 5 μm); acetonitrile-0.1% phosphoric acid solution (11:89) as the mobile phase; and a detection wavelength of 324 nm. The theoretical plate number, calculated based on the peak of morin A, should be no less than 8000.

[0026] To prepare the reference solution, accurately weigh an appropriate amount of morin A reference standard, add 50% methanol to prepare a solution containing 80 μg per ml.

[0027] To prepare 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 50 ml of 50% methanol, stopper tightly, weigh it, sonicate (power 600W, frequency 40kHz) for 20 minutes, cool it, weigh it again, make up the lost weight with 50% methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0028] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0029] Table 1 shows the calculation of the transfer rate of quercetin in the sample solution:

[0030] This product contains 49.4 mg to 127.9 mg of morin A per 1g.

[0031] The transfer rate of morin A content in the decoction of medicinal materials ranged from 34.4% to 63.9%.

[0032] The characteristic chromatogram was determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0033] 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% formic acid solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the detection wavelength was 280 nm. The theoretical plate number, calculated based on the peak of morin A, should not be less than 8000.

[0034] Table 2 shows the gradient elution table for chromatographic conditions and system suitability testing:

[0035]

[0036] The preparation of the reference solution included: taking approximately 0.25g of mulberry bark reference material, placing it in a stoppered conical flask, adding 50ml of 60% methanol, sealing tightly, sonicating (600W power, 40kHz frequency) for 20 minutes, cooling, shaking well, filtering, and collecting the filtrate as the reference solution. Separately, taking appropriate amounts of morin A and morin G reference standards, adding 50% methanol to prepare solutions containing 80μg each per ml, as reference solutions.

[0037] The preparation of the test solution includes: taking an appropriate amount of this product, grinding it into a fine powder, and taking about 0.1 g, preparing the test solution in the same way as the preparation method of the reference solution of the control medicinal material.

[0038] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0039] like Figure 2 As shown, 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 5 (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 2, 3, and 8 should be within ±4% of the corresponding peaks in the reference medicinal material, and the retention times of characteristic peaks 4, 6, and 7 should be within ±1% of the corresponding peaks in the reference medicinal material. Figure 2 In the sample, peak 1 represents morin A; peak 5 (S) represents morin G; each milligram of the test sample is approximately equivalent to 0.012 grams of medicinal slices.

[0040] In this plan, the selection of raw materials is based on 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. Mulberry bark is a cultivated product, distributed throughout my country. It is mainly produced in Sichuan, Guizhou, Guangxi, and Hubei provinces, where its cultivation has reached a large scale and is the main source of commercial mulberry bark. 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 are Jingyan County (Sichuan), Zhongjiang County (Sichuan), Yashan County (Guizhou), Anshun County (Guizhou), Yunlin County (Guangxi), and Enshi Prefecture (Hubei). Ultimately, 18 qualified batches were selected for research on standard decoctions.

[0041] Table 3 shows the transfer rate of 18 batches of mulberry bark standard decoction:

[0042]

[0043]

[0044] In the table above, based on the content of mulberry bark slices, standard decoction content, and extract yield data from 18 batches, the average transfer rate of standard decoction content was calculated to be 49.2%. The range calculated using the mean plus or minus 3 times the SD is 37.0% to 61.3%. The range calculated using the mean of 70% to 130% is 34.4% to 63.9%. Considering the actual range of 44.8% to 56.7%, 34.4% to 63.9% was taken as the upper and lower limits of the standard decoction transfer rate.

[0045] Modern research shows that mulberry bark contains various components, such as Diels-Alder adducts, flavonoids (including flavonoids, dihydroflavonoids, and chalcones), furans, coumarins, terpenes, sterols, sugars, volatile oils, and rutin (moriposide). Mulberry bark possesses a variety of pharmacological activities, including hypoglycemic and hypotensive effects, bronchodilatory and diuretic effects, analgesic and anti-inflammatory effects, hepatoprotective effects, antiviral effects, and antitumor effects. The 2020 edition of the Chinese Pharmacopoeia lists morin A as an indicator component for quality control of mulberry bark; therefore, this study selected morin A as the quality control indicator component for the standard decoction of mulberry bark.

[0046] Among them, based on the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules", and combined with the data of 18 batches of mulberry bark standard decoction, the content limits and upper and lower limits of the standard decoction were calculated.

[0047] The range calculated using the mean plus or minus 3 times the SD is: 49.4 mg to 127.9 mg;

[0048] The range calculated using 70%–130% of the mean is 62.0 mg–115.2 mg.

[0049] Based on the actual range of 75.2mg to 118.7mg, the average value of 70% to 130%, i.e., 49.4mg to 127.9mg, was adopted as the upper and lower limits of the standard decoction content.

[0050] Based on the content data of 18 batches of mulberry bark slices, standard decoction content, and extract yield, the average transfer rate of the standard decoction content was calculated to be 49.2%. The range calculated using the mean plus or minus 3 times the SD was 37.0%–61.3%. The range calculated using the mean of 70%–130% was 34.4%–63.9%. Considering the actual range of 44.8%–56.7%, 34.4%–63.9% was taken as the upper and lower limits of the standard decoction transfer rate.

[0051] The research and standardization of characteristic maps are as follows:

[0052] The chromatogram of the standard decoction of mulberry bark with experimental batch number BT(SBP)220101 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 mulberry senna G was selected as the reference peak. The relative retention times of the characteristic peaks in the chromatogram were calculated.

[0053] like Figure 3 As shown; by comparing the chromatogram with that of the reference standard, it can be seen that among the five characteristic peaks in the characteristic chromatogram of the mulberry bark standard decoction, peak 5 (S) is morin A. According to the "Quality Standard of Mulberry Bark Formula Granules" YBZ-PFKL-2021106, peak 1 is morin A and peak 5 is morin G.

[0054] Table 4 shows a comparison of the retention times of six batches of mulberry bark standard decoction:

[0055]

[0056]

[0057] Table 5 shows the relative retention times of six batches of mulberry bark standard decoction:

[0058]

[0059]

[0060] The results showed that the characteristic chromatograms of the six batches (one batch randomly selected from each origin) of mulberry bark standard decoction all showed chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the control herb. 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. (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.)

[0061] The national standard "Quality Standard for Mulberry Bark Formula Granules" (YBZ-PFKL-2021106) stipulates that five characteristic peaks should correspond to the retention times of the reference medicinal material peaks. Specifically, two peaks (peak 1: morin A, peak 5: morin G) should correspond to the retention times of their respective reference medicinal material peaks, and the relative retention times of peaks 2-4 are also specified. Based on the results of six batches of standard mulberry bark decoctions and the retention time data from the repeatability and precision tests, it can be seen that the retention times of the five characteristic peaks are stable and correspond to the retention times of the reference medicinal material peaks. The relative peak drift of peaks 2-4 is very small, basically within ±8%, indicating that the retention times of each characteristic peak are stable. Therefore, the regulations stipulate that the chromatogram of the test sample should show 5 characteristic peaks, and the retention times should correspond to the 5 characteristic peaks in the chromatogram of the reference medicinal material. Two of these peaks should correspond to the retention times of the corresponding reference material peaks, and the peak corresponding to the peak of the Sanghuang ketone G reference material is designated as the S peak. The relative retention times of each characteristic peak and the S peak should be calculated, and these relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.420 (peak 2), 0.438 (peak 3), and 0.550 (peak 4).

[0062] In this plan, the research and standard setting of leachate are as follows:

[0063] Grind the product into a fine powder, accurately weigh approximately 2g, and precisely add 100ml of ethanol. Determine the soluble extract using the hot extraction method as described in the General Chapter 2201 of the Chinese Pharmacopoeia 2020. The test results for 18 batches of standard Platycladus orientalis leaf decoction are shown in the table below.

[0064] Table 6 is a comparison table of the extract results of 18 batches of mulberry bark:

[0065]

[0066] The average value of the alcohol-soluble extract from 18 batches of standard decoctions was 32.7%.

[0067] The range calculated using the mean plus or minus 3 times the SD is: 25.0% to 40.3%;

[0068] The range calculated using 70% to 130% of the mean is 22.9% to 42.5%.

[0069] Based on the actual range, it is 29.7% to 38.4%;

[0070] The upper and lower limits of the extract in the standard decoction were set at 25.0% to 40.3%, with the mean value plus or minus 3 times the SD. In this invention, taking mulberry bark decoction as an example, the key quality indicators such as the main content, characteristic chromatogram, and extract of mulberry bark after decoction preparation were specified, and the transfer rate was investigated. The range of transfer rate of the main content of mulberry bark was specified, ensuring the effectiveness of the mulberry bark decoction after following the standard preparation method.

[0071] 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, 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.

[0072] 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 the standard decoction of mulberry bark, characterized in that, The preparation process includes the following steps: S1: Weigh out the mulberry bark slices, place them in a clay pot, 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 to obtain a mixed medicinal liquid. S2: Add 8 times the amount of water again, bring to a boil over high heat, then simmer over low heat for 20 minutes to obtain a further mixed medicinal liquid; S3: Based on the above steps, filter the mixed solution through a 200-mesh filter, cool the filtrate to room temperature in a cold water bath, and combine the two filtrates; S4: Vacuum concentration is carried out at a temperature of 50℃ until the weight ratio of medicinal material to fluid extract is 1:

1. The concentrated liquid is then collected and freeze-dried to obtain the sample.

2. The preparation process of the standard mulberry bark decoction according to claim 1, characterized in that, In S4, vacuum concentration is performed at a temperature of 50°C, with an absolute vacuum of 30-40 mbar.

3. The preparation process of the mulberry bark standard decoction according to claim 1, characterized in that, In step S4, the extraction rate and extract indicators of mulberry bark in the decoction are measured through concentration and freeze-drying operations.

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