Preparation process of honey white mulberry root-bark standard decoction

Abstract

The invention discloses a preparation technology of a honey 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 honey 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 honey white mulberry root-bark standard decoction, key quality indexes such as the main content, the characteristic chromatogram and the extract of the honey 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 honey mulberry bark standard decoction. Background Technology

[0002] Mulberry bark is the dried root bark of the dicotyledonous plant Moraceae after removing the cork. 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. Honey-processed mulberry bark is a processed traditional Chinese medicine obtained by processing mulberry bark with honey. Honey-processed mulberry bark has a dark outer surface and a sweet taste. It is milder in medicinal properties and has higher medicinal value than natural mulberry bark, possessing the effects of clearing heat and moistening the lungs, promoting diuresis and reducing swelling, protecting the liver, clearing heat and removing fire, and preventing hypertension. Traditional Chinese medicine decoctions are a traditional and widely used form of medication in clinical practice. Standard decoctions are single-herb decoctions prepared using standardized processes based on traditional Chinese medicine theory and 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 standards for the standard decoction of honey-mulberry bark to measure its quality and efficacy 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 honey-mulberry bark. Summary of the Invention

[0004] The main objective of this invention is to provide a preparation process for a standard decoction of honey-processed 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 the standard decoction of honey-mulberry bark includes the following steps:

[0006] S1: Weigh out the slices of honey-processed mulberry bark, 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 S4, vacuum concentration is performed at a temperature of 50°C, with an absolute vacuum of 30-40 mbar.

[0011] In step S4, the extraction rate and extract indicators of mulberry bark in the decoction are measured through concentration and freeze-drying operations.

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

[0013] The present invention discloses a preparation process for a standard decoction of honey mulberry bark. It specifies key quality indicators such as the main content, characteristic spectrum, and extract of honey mulberry bark after decoction preparation, and examines the transfer rate. By specifying the range of transfer rate of the main content of honey mulberry bark, the effectiveness of the decoction is guaranteed after following the standard preparation 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 honey-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 honey 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 The preparation process of the standard decoction of honey-processed mulberry bark is as follows: Weigh an appropriate amount of honey-processed 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, 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, cool the filtrate to room temperature in a cold water bath, combine the two filtrates, and concentrate under vacuum at 50℃ and an absolute vacuum of 30–40 mbar until the weight ratio of the medicinal material to the fluid extract is approximately 1:1. Collect the concentrated liquid and freeze-dry to obtain the sample. Subsequent concentration and freeze-drying are used to measure the yield and extract content of the traditional Chinese medicine in the decoction.

[0020] The aqueous extract of mulberry bark 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: 21.6% to 29.7%.

[0023] The 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 31.7%–58.9%.

[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 morin in the sample solution:

[0030]

[0031] This product contains 19.5 mg to 48.6 mg of morin A per 1g.

[0032] The transfer rate of morin A content in the decoction of medicinal materials ranged from 21.3% to 43.9%.

[0033] The characteristic chromatogram was determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0034] 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.

[0035] Table 2 shows the gradient elution table for chromatographic conditions and system suitability testing:

[0036]

[0037] 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 of each per ml, as reference solutions.

[0038] 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.

[0039] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0040] like Figure 2 As shown, the chromatogram of the test sample should show 7 characteristic peaks, which should correspond to the retention times of the 7 characteristic peaks in the chromatogram of the reference decoction pieces. Two of these peaks should correspond to the retention times of the corresponding reference peaks. The peak corresponding to the peak of mulberry ginseng G is the S peak. Calculate the relative retention times of each characteristic peak and the S peak; the relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.10 (peak 1), 0.13 (peak 2), 0.42 (peak 4), 0.44 (peak 5), and 0.55 (peak 6); where peak 3 represents morin A; peak 7 (S) represents mulberry ginseng G; each milligram of the test sample is approximately equivalent to 0.078 grams of decoction pieces.

[0041] 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. Honey 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 honey 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.

[0042] Table 3 shows the transfer rate of 18 batches of standard decoction of honey-processed mulberry bark:

[0043]

[0044]

[0045] In the table above, based on the content of 18 batches of honey mulberry bark slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 32.6%. The range calculated using the mean plus or minus 3 times the SD is 21.3% to 43.9%. The range calculated using the mean of 70% to 130% is 22.8% to 42.4%. Considering the actual range of 27.8% to 39.2%, 21.3% to 43.9% is taken as the upper and lower limits of the standard decoction transfer rate.

[0046] 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.

[0047] 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 decoction of honey mulberry bark, the content limits and upper and lower limits of the standard decoction were calculated. The range calculated using the mean plus or minus 3 times the SD is 19.5mg to 48.6mg; the range calculated using 70% to 130% of the mean is 23.8mg to 44.3mg.

[0048] Based on the actual range of 25.2mg to 42.5mg, the upper and lower limits of the standard decoction content were set as the mean plus or minus 3 times the SD, i.e., 19.5mg to 48.6mg.

[0049] Based on the content data of 18 batches of honey mulberry bark slices, standard decoction content, and extract yield, the average transfer rate of the standard decoction content was calculated to be 32.6%. The range calculated using the mean plus or minus 3 times the SD was 21.3%–43.9%. The range calculated using the mean of 70%–130% was 22.8%–42.4%. Considering the actual range of 27.8%–39.2%, 21.3%–43.9% was taken as the upper and lower limits of the standard decoction transfer rate.

[0050] The research and standardization of characteristic maps are as follows:

[0051] like Figure 3 As shown, the chromatogram of the standard decoction of honey-processed mulberry bark with experimental batch number BT(MSBP)220101 was selected as the reference chromatogram to establish a control characteristic chromatogram. Based on the data analysis results, seven 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.

[0052] By comparing the chromatogram with that of the reference standard, it can be seen that among the seven characteristic peaks in the characteristic chromatogram of the honey mulberry bark standard decoction, peak 5 (S) is morin A; according to the "Quality Standard of Honey Mulberry Bark Formula Granules" YBZ-PFKL-2021093, peak 3 is morin A and peak 7 is morin G; wherein, peak 3: morin A; peak 7 (S): morin G; wherein, peak 3: morin A; peak 7 (S): morin G.

[0053] Table 4 shows a comparison of the retention times of six batches of standard decoction of honey-processed mulberry bark:

[0054]

[0055]

[0056] Storage time of honey mulberry bark slices

[0057]

[0058] Table 5 shows the relative retention schedule for six batches of standard decoction of honey mulberry bark:

[0059]

[0060]

[0061] Relative shelf life of honey mulberry bark slices

[0062]

[0063] The results showed that the characteristic chromatograms of the six batches of honey mulberry bark standard decoction samples all showed chromatographic peaks corresponding to the retention times of the seven main characteristic peaks in the control medicinal material. The relative retention time RSD of peaks 1-7 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 in accordance with the batch of medicinal slices to prove that the prepared sample had almost no effect on the characteristic chromatogram of the medicinal slices.)

[0064] The national standard YBZ-PFKL-2021093, "Quality Standard for Mulberry Bark Formula Granules," stipulates that the retention times of seven characteristic peaks should correspond to those of the reference medicinal material peaks. Specifically, two peaks (peak 3: morin A, peak 7: morin G) should correspond to the retention times of their respective reference medicinal material peaks. The relative retention times of peaks 1, 2, and 4-6 are also specified. Based on the results of six batches of standard mulberry bark decoctions and the retention time data from repeatability and precision tests in the methodology, it can be seen that the retention times of the seven characteristic peaks are stable and correspond to those of the reference medicinal material peaks. The relative peak drift of peaks 1, 2, and 4-6 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 7 characteristic peaks, and the retention times should correspond to the 7 characteristic peaks in the chromatogram of the reference decoction pieces. Two of these peaks should correspond to the retention times of the corresponding reference peaks, and the peak corresponding to the peak of the Sanghuang ketone G reference 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: 0.10 (peak 1), 0.13 (peak 2), 0.42 (peak 4), 0.44 (peak 5), and 0.55 (peak 6).

[0065] In this plan, the research and standard setting of leachate are as follows:

[0066] Grind the product into a fine powder, take approximately 2g, accurately weigh it, and accurately 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 decoction of honey-processed mulberry bark are shown in the table below.

[0067] Table 6 is a comparison table of the extract results of 18 batches of honey mulberry bark:

[0068]

[0069]

[0070] The average value of the alcohol-soluble extract from 18 batches of standard decoctions was 40.7%.

[0071] The range calculated using the mean plus or minus 3 times the SD is 35.8% to 45.7%.

[0072] The range calculated using 70% to 130% of the mean is 28.5% to 52.9%.

[0073] Based on the actual range of 38.9% to 44.1%;

[0074] The upper and lower limits of the standard decoction extract were taken as the mean value plus or minus 3 times the SD value, which ranged from 31.7% to 58.9%.

[0075] In this invention, taking honey mulberry bark decoction as an example, the key quality indicators such as the main content, characteristic spectrum and extract of honey mulberry bark after decoction preparation are specified, and the transfer rate is investigated. The range of the transfer rate of the main content of honey mulberry bark is specified to maintain the effectiveness of honey mulberry bark decoction after preparation according to the standard method.

[0076] 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.

[0077] 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 honey-mulberry bark, characterized in that, The preparation process includes the following steps: S1: Weigh out the slices of honey-processed mulberry bark, 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 decoction of honey-processed mulberry bark 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 standard decoction of honey-processed mulberry bark 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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