Preparation process of standard fructus perillae decoction

Abstract

The invention discloses a preparation technology of a standard fructus perillae decoction, and relates to the technical field of traditional Chinese medicine decoction treatment.The preparation technology comprises the steps that fructus perillae decoction pieces are weighed and placed in a marmite to be decocted twice; comprising the following steps: Step 1: adding water which is 8 times of fructus perillae decoction pieces into a marmite for the first time, soaking for 30 minutes, boiling with strong fire, and decocting with slow fire for 30 minutes; step 2, adding water which is 6 times of the fructus perillae decoction pieces into the marmite for the second time, boiling with strong fire, and decocting with slow fire for 20 minutes; 3, filtering the liquid medicine with 200 meshes, and cooling the filtrate to room temperature in a cold water bath; step 4, combining the two filtrates, and carrying out vacuum concentration at the temperature of 50 DEG C; and Step 5, concentrating until the weight ratio of the medicinal material to the fluid extract is 1: 1, collecting a concentrated solution, and freeze-drying to obtain a sample. According to the method, the component transfer condition of the fructus perillae decoction pieces to the decoction is measured through objective quality standards, and the quality degree and effectiveness of the decoction are ensured.

Description

Technical Field

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

[0002] Perilla seeds are the dried, ripe fruit of the perilla plant (Perilla frutescens), belonging to the Lamiaceae family. It is mostly cultivated. Mainly produced in Gansu, Hubei, Jiangsu, Hunan, Zhejiang, Anhui, and Henan provinces, perilla seeds grow in wetlands, roadsides, villages, and wastelands, or are cultivated. It prefers a warm, humid climate and thrives best in loose, fertile, and sunny locations. It has a pungent taste and warm properties. It enters the lung meridian. Its functions include lowering qi and resolving phlegm, relieving cough and asthma, and moistening the intestines. Clinically, it is known as perilla seed, stir-fried perilla seed, honey-processed perilla seed, and perilla seed frost. The plant typically grows to over 1 meter tall, with green or purple, square-columnar stems with four blunt ridges, covered in long, soft hairs. Leaves opposite, herbaceous, broadly ovate or nearly orbicular, 7 to 13 cm long, 4.5 to 10 cm wide, apex shortly acuminate or abruptly acuminate, base broadly cuneate to rounded, margin with coarsely serrated, lacerate margins, purple on both sides or only purple below, purplish-green above, sparsely pubescent, below appressed pubescent. Petioles 3 to 5 cm long. Flowers bloom in autumn, purplish-red, arranged in axillary, densely flowered, lateral racemes; bracts broadly ovate or nearly orbicular, about 4 mm long, with reddish-brown glandular dots, glabrous; calyx campanulate, straight, about 3 mm long, 10-veined, pubescent and glandular-dotted, limb bilabiate, upper lip 3-lobed, middle lobe smaller, lower lip 2-lobed, longer than the upper lip; corolla 3 to 4 mm long, corolla tube short, throat nearly campanulate, limb nearly bilabiate, upper lip slightly notched, lower lip 3-lobed, middle lobe particularly large; stamens 4, nearly as high as the corolla lobes. The calyx is significantly enlarged, 1.1 cm long, horizontal or drooping, with one side of the base swollen. The nutlet is spherical, 1.5 mm in diameter, brown, and reticulate.

[0003] 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 perilla seed decoction as an example. It specifies key quality indicators such as the main content, characteristic chromatograms, and extracts of perilla seed after decoction preparation, examines the transfer rate, and defines the range of transfer rates for the main content of perilla seed. This ensures the effectiveness of perilla seed decoction after preparation according to standard methods. Summary of the Invention

[0004] The main objective of this invention is to provide a preparation process for a standard decoction of perilla seeds, in order to solve the problems mentioned in the background art.

[0005] In order to derive the preparation process of the standard decoction of perilla seeds, the present invention conducted relevant pharmaceutical research, such as screening 15 batches of perilla seed medicinal materials from major producing areas across the country for quality comparison research, and preparing 15 batches of standard decoctions in accordance with the "Management Standards for Traditional Chinese Medicine Decoction Rooms in Medical Institutions".

[0006] To achieve the above objectives, the technical solution adopted by this invention is as follows: The preparation process of the standard decoction of perilla seeds involves first weighing perilla seed slices, placing them in a clay pot, and decocting them twice; including the following steps:

[0007] Step 1: Add 8 times the amount of water as perilla seed slices to the clay pot for the first time, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes.

[0008] Step 2: Add 6 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 minutes.

[0009] Step 3: Then filter the above medicine solution through a 200-mesh filter and cool the filtrate to room temperature in a cold water bath;

[0010] Step 4: Combine the two filtrates and concentrate them under vacuum at 50°C;

[0011] Step 5: Concentrate the extract to a 1:1 weight ratio of medicinal material to fluid extract, collect the concentrate, and freeze-dry to obtain the sample.

[0012] Preferably, in Step 4, vacuum concentration is performed under an absolute vacuum of 30–40 mbar.

[0013] The present invention has the following beneficial effects:

[0014] The preparation process of the standard decoction of perilla seeds in this invention has objective quality standards to measure the component transfer of perilla seed slices into decoction, and to ensure the quality and effectiveness of the decoction.

[0015] 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

[0016] Figure 1 This is a flowchart illustrating the preparation process of the standard perilla seed decoction of the present invention.

[0017] Figure 2 This is a comparative characteristic chromatogram of the test solution of the standard decoction of perilla seeds of the present invention;

[0018] Figure 3 This is a comparative characteristic spectrum of the standard decoction of perilla seeds in this invention. Detailed Implementation

[0019] 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.

[0020] Example 1:

[0021] like Figure 1 As shown: The preparation process of the standard decoction of perilla seeds involves first weighing the perilla seed slices, placing them in a clay pot, and decocting them twice, including the following steps:

[0022] Step 1: First, add 8 times the amount of water (perilla seed slices) to the clay pot, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes. Step 2: Second, add 6 times the amount of water (perilla seed slices) to the clay pot, bring to a boil over high heat, then simmer over low heat for 20 minutes. Step 3: Then filter the above decoction 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 50°C. Step 5: Concentrate until the weight ratio of medicinal material to fluid extract is 1:1, collect the concentrate, and freeze-dry to obtain the sample.

[0023] Extract yield: After vacuum concentration of the perilla seed aqueous extract, the concentrate is freeze-dried to dryness. The dried extract powder is collected and weighed to calculate the extract yield. The formula for extract yield is as follows:

[0024]

[0025] Ointment yield range: 5.0% to 7.9%.

[0026] 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 11.8%–21.4%.

[0027] Content determination

[0028] Rosmarinic acid was determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020).

[0029] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; methanol-0.1% formic acid solution (40:60) was used as the mobile phase; the detection wavelength was 330 nm. The theoretical plate number, calculated based on the rosmarinic acid peak, should be no less than 3000.

[0030] Preparation of reference solution: Take an appropriate amount of rosmarinic acid reference standard, accurately weigh it, and add methanol to prepare a solution containing 80 μg per ml.

[0031] 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, accurately add 50ml of 80% methanol, stopper tightly, weigh it, sonicate (power 600W, frequency 40kHz) for 20 minutes, cool it, weigh it again, make up the lost weight with 80% methanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0032] Determination method: Accurately pipette 5 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0033] Table 1 shows the transfer rate of rosmarinic acid content from medicinal materials to the standard decoction:

[0034]

[0035] This product contains 6.8mg to 19.2mg of rosmarinic acid (C18H16O8) per 1g.

[0036] The transfer rate of rosmarinic acid content in the decoction of medicinal materials ranged from 14.9% to 27.7%.

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

[0038] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the 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 284 nm. The theoretical plate number, calculated based on the rosmarinic acid peak, should be no less than 3000.

[0039] Table 2 shows the gradient elution table:

[0040]

[0041] Preparation of reference solution: Take 1g of Perilla frutescens seed reference material, place it in a stoppered conical flask, add 25ml of 80% methanol, seal tightly, sonicate (600W power, 40kHz frequency) for 30 minutes, shake well, filter, and take the filtrate as the reference solution. Separately, take an appropriate amount of rosmarinic acid reference standard, add methanol to prepare a solution containing 80μg per ml, and use this as the reference solution.

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

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

[0044] The chromatogram of the test sample should show 6 characteristic peaks, and the retention times should correspond to the 6 characteristic peaks in the chromatogram of the reference herb. The peak corresponding to the rosmarinic acid reference peak is designated as the S peak. The relative retention times of each characteristic peak and the S peak should be calculated, and the relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.341 (peak 1), 0.381 (peak 2), 0.457 (peak 3), 0.716 (peak 4), and 0.938 (peak 5).

[0045] Example 2:

[0046] 1. Raw material selection

[0047] 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. Perilla seeds are a cultivated product, distributed throughout my country. They are mainly produced in Shaanxi, Sichuan, Gansu, and Henan provinces, where cultivation has reached a large scale and is a major source of commercial perilla seeds. Based on a comprehensive evaluation of raw material quality and sustainable resource supply, the preliminary producing areas for large-scale production were determined to be Bin County in Xianyang City, Shaanxi Province; Dazhu County in Dazhou City, Sichuan Province; Ning County in Qingyang City, Gansu Province; Guangshan County in Xinyang City, Henan Province; and Tong County in Nanyang City, Henan Province. Ultimately, 15 qualified batches were selected for research on standard decoctions.

[0048] 1. Research and standard setting on extract yield and transfer rate

[0049] Table 3 shows the transfer rate (rosmarinic acid) of 15 batches of standard perilla seed decoction.

[0050]

[0051]

[0052] Based on the content of 15 batches of perilla seed slices, the content of standard decoction (rosmarinic acid), and the yield of extract, the average transfer rate of standard decoction content was calculated to be 16.5%.

[0053] The range calculated using the mean plus or minus 3 times the SD is 16.0% to 26.6%.

[0054] The range calculated using 70% to 130% of the mean is 14.9% to 27.7%.

[0055] Based on the actual range, it is 18.6% to 24.2%;

[0056] It is recommended to take 14.9% to 27.7% as the upper and lower limits of the standard decoction transfer rate.

[0057] 2. Content research and standard setting

[0058] Modern research shows that perilla seeds contain various components, such as fatty oils, amino acids, trace elements, apigenin, luteolin, and flavonoids. Perilla seeds possess a variety of pharmacological activities, including lowering blood lipids, improving memory, relieving cough and asthma, anti-aging, and anti-allergy. The 2020 edition of the Chinese Pharmacopoeia lists rosmarinic acid as an indicator component for the quality control of perilla seeds. Combined with the provisions under the "Content Determination" section of the National Drug Standard YBZ-PFKL-2021159 for Perilla Seed Formula Granules, this study selected rosmarinic acid as the quality control indicator component for the standard perilla seed decoction.

[0059] Based on the "Technical Requirements for Quality Control and Standard Formulation of Traditional Chinese Medicine Formula Granules", and combined with data from 15 batches of standard decoction of Perilla Seed, the limits and upper and lower limits of rosmarinic acid content in the standard decoction were calculated.

[0060] The range calculated using the mean plus or minus 3 times the SD is 6.8 mg to 19.2 mg.

[0061] The range calculated using 70%–130% of the mean is 9.1 mg–16.9 mg.

[0062] Based on the actual range of 10.4mg to 16.8mg, it is recommended to use the mean plus or minus 3 times the SD, i.e., 6.8mg to 19.2mg, as the upper and lower limits of the standard decoction content.

[0063] Based on the content of 15 batches of perilla seed slices, the content of standard decoction (rosmarinic acid), and the yield of extract, the average transfer rate of standard decoction content was calculated to be 16.5%.

[0064] The range calculated using the mean plus or minus 3 times the SD is 16.0% to 26.6%.

[0065] The range calculated using 70% to 130% of the mean is 14.9% to 27.7%.

[0066] Based on the actual range, it is 18.6% to 24.2%;

[0067] It is recommended to take 14.9% to 27.7% as the upper and lower limits of the standard decoction transfer rate.

[0068] 4. Characteristic spectrum research and standard setting

[0069] The standard decoction of Perilla frutescens seeds with batch number BT(ZSZ)220201 was selected as the reference chromatogram to establish a control characteristic chromatogram. Based on the data analysis results, six common peaks were finally selected as characteristic peaks. The generated standard chromatogram is shown below. Figure 3 Rosmarinic acid was selected as the reference peak, and the relative retention times of the characteristic peaks in the chromatogram were calculated.

[0070] Comparison with the chromatogram of the reference standard revealed that among the six characteristic peaks in the chromatogram of the Perilla Seed Standard Decoction, peak 6(S) was rosmarinic acid. This chromatogram conforms to the national drug standard for Perilla Seed Granules.

[0071] According to YBZ-PFKL-2021159, peak 3 is caffeic acid; peak 6 (S) is rosmarinic acid.

[0072] Table 4 shows the relative retention times of 15 batches of standard Perilla Seed decoction.

[0073]

[0074]

[0075] The results showed that the chromatograms of five random batches of Perilla Seed standard decoction samples and processed medicinal materials all exhibited chromatographic peaks corresponding to the retention times of the six main characteristic peaks in the control medicinal materials. The relative retention times (RSD) of peaks 1-5 were less than 2%, indicating that the elution times of each peak were consistent across batches, and the characteristic chromatograms were not affected by the process. (Note: The batch number of the standard decoction was selected to correspond to the batch of processed medicinal materials to demonstrate that the prepared sample had almost no effect on the characteristic chromatograms of the processed medicinal materials.)

[0076] The national standard YBZ-PFKL-2021159, "Perilla Seed Formula Granules," only specifies the retention times of peak 6 and the reference standard, and the relative retention times of peaks 1-5 and peak S. Combining the results of five random batches of Polygonum multiflorum standard decoction and the retention time data from the repeatability and precision tests in the methodology, it can be seen that the retention times of peak 6 and the reference standard are corresponding, and the relative peak drift of peaks 1-5 and peak S is very small, basically within ±10%. Therefore, it is stipulated that the chromatogram of the test sample should show six characteristic peaks, corresponding to the retention times of the six characteristic peaks in the chromatogram of the reference medicinal material. The peak corresponding to the rosmarinic acid reference peak is the S peak. The relative retention times of each characteristic peak and peak S should be calculated, and their relative retention times should be within ±10% of the specified values. The specified values ​​are: 0.341 (peak 1), 0.381 (peak 2), 0.457 (peak 3), 0.716 (peak 4), and 0.938 (peak 5).

[0077] 5. Leachate research and standard setting

[0078] 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 15 batches of Perilla Seed standard decoction are shown in Table 5.

[0079] Table 5 shows the results of extract determination of 15 batches of perilla seeds.

[0080]

[0081]

[0082] The average value of the alcohol-soluble extract from 15 batches of standard decoction was 16.5%.

[0083] The range calculated using the mean plus or minus 3 times the SD is 11.8% to 21.2%.

[0084] The range calculated using 70% to 130% of the mean is 11.5% to 21.4%.

[0085] Based on the actual range, it is 14.7% to 19.6%;

[0086] It is recommended to take 11.8% to 21.4% as the upper and lower limits of the standard decoction extract.

[0087] 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.

[0088] 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 perilla seeds, firstly, weigh the perilla seed slices, place them in a clay pot, and decoct twice, characterized in that... Includes the following steps: Step 1: Add 8 times the amount of water as perilla seed slices to the clay pot for the first time, soak for 30 minutes, bring to a boil over high heat, then simmer over low heat for 30 minutes. Step 2: Add 6 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 minutes. Step 3: Then filter the above medicine 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 50°C; Step 5: Concentrate the extract to a 1:1 weight ratio of medicinal material to fluid extract, collect the concentrate, and freeze-dry to obtain the sample.

2. The preparation process of the standard perilla seed 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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