Preparation process of salted semen cuscutae standard decoction

Abstract

The invention discloses a preparation technology of a salted semen cuscutae standard decoction, and relates to the technical field of traditional Chinese medicine decoction treatment.The preparation technology comprises the following steps that firstly, a proper amount of salted semen cuscutae decoction pieces are weighed, placed in an earthen pot and decocted twice, and the preparation technology comprises the following steps that firstly, water 8 times of the salted semen cuscutae decoction pieces is added into the earthen pot for the first time, soaked for 30 min, boiled with strong fire and then decocted with slow fire for 30 min; step 2, adding water which is 6 times of the salt semen cuscutae 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 salted semen cuscutae 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 Cuscuta chinensis with salt. Background Technology

[0002] Dodder, an annual parasitic herb belonging to the genus *Cuscuta* in the Convolvulaceae family, has slender, twining, yellow stems; it is leafless; its bracts are small and scale-like; its flowers grow in clusters, with stout pedicels; the calyx is cup-shaped with ovate or oblong lobes; the corolla is white, urn-shaped or bell-shaped; the capsule is nearly spherical; the seeds are ovoid, light brown, and have a rough surface; it flowers from July to August and fruits from August to September. The name *Cuscuta* actually means "silk-spinning seed," referring to the seed coat cracking after boiling to reveal a small, yellowish-white embryo, resembling the spinning of silk threads. Dodder is distributed in Asia, Africa, and Australia, and in most parts of China; it prefers hot, humid climates and grows mostly at altitudes of 200-3000 meters in fields, sunny slopes, roadside thickets, or coastal dunes; it typically parasitizes various plants in the Fabaceae, Asteraceae, and Zygophyllaceae families. Dodder primarily reproduces and spreads through seeds. The dried, mature seeds of dodder are used medicinally, a practice first recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica), which states that they "treat injuries, replenish deficiencies, boost energy, and promote longevity." As both food and medicine, they can also treat excessive urination or incontinence, tonify the kidneys and replenish essence, and nourish the liver and improve eyesight. Modern research indicates that dodder enhances immunity and improves blood circulation. However, dodder is a harmful weed in soybean-producing areas. It has a very wide host tolerance, with a single plant capable of parasitizing multiple hosts. If any host is infected with a virus, the virus can be transmitted to other hosts via dodder, thus harming crops such as sesame, ramie, peanuts, and potatoes.

[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 salt-cured dodder seed decoction as an example. It specifies key quality indicators such as the main content, characteristic chromatograms, and extracts of salt-cured dodder seed after decoction preparation, examines the transfer rate, and defines the range of transfer rates for the main content of salt-cured dodder seed. This ensures the effectiveness of salt-cured dodder seed decoction after standard preparation. Summary of the Invention

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

[0005] In order to derive the preparation process of the standard decoction of Cuscuta chinensis, this invention conducted relevant pharmaceutical research, such as screening 15 batches of Cuscuta chinensis 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 salted dodder seed involves first weighing an appropriate amount of salted dodder 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 salted dodder 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 as salted dodder seed slices 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 salted dodder seed of the present invention has objective quality standards to measure the component transfer of salted dodder 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 decoction of salted dodder seed according to the present invention.

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

[0018] Figure 3 This is a comparative characteristic chromatogram of the standard decoction of salted dodder seed of the present 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 salted dodder seed involves first weighing an appropriate amount of salted dodder seed slices, placing them in a clay pot, and decocting them twice, including the following steps:

[0022] Step 1: Add 8 times the amount of water as salted dodder 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.

[0023] Step 2: Add 6 times the amount of water as salted dodder seed slices to the clay pot for the second time, bring to a boil over high heat, then simmer over low heat for 20 minutes.

[0024] 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;

[0025] Step 4: Combine the two filtrates and concentrate them under vacuum at 50°C;

[0026] 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.

[0027] Extract yield: After vacuum concentration of the aqueous extract of Cuscuta chinensis (Southern Cuscuta chinensis), 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:

[0028]

[0029] Ointment yield range: 13%–20%.

[0030] Extractives: Determined by hot extraction method according to the alcohol-soluble extractives determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 2201) using ethanol as solvent. The extractives range of the standard decoction is 17.9%–40.6%.

[0031] Content determination

[0032] Hyperoside was determined by high performance liquid chromatography (General Chapter 0512, Chinese Pharmacopoeia 2020).

[0033] Chromatographic conditions and system suitability test: Octadecylsilane-bonded silica gel was used as the stationary phase; acetonitrile-0.1% phosphoric acid solution (17:83) was used as the mobile phase; the detection wavelength was 360 nm. The theoretical plate number, calculated based on the hyperoside peak, should not be less than 5000.

[0034] Preparation of reference solution: Take an appropriate amount of hyperoside reference standard, accurately weigh it, and add methanol to prepare a solution containing 50 μg per ml.

[0035] Preparation of the test solution: Take an appropriate amount of this product, grind it into a fine powder, take about 0.2g, weigh it accurately, place it in a stoppered conical flask, accurately add 25ml of 70% ethanol, stopper tightly, weigh it, sonicate (power 300W, frequency 40kHz) for 30 minutes, cool it, weigh it again, make up the weight loss with 70% ethanol, shake well, filter it, and take the filtrate to obtain the test solution.

[0036] Determination method: Accurately pipette 10 μl of the reference solution and the test solution into the liquid chromatograph and determine the result.

[0037] Table 1 shows the hyperoside content transfer rate from the medicinal materials to the standard decoction:

[0038]

[0039] This product contains 4.7 mg / g to 8.7 mg / g of hyperoside per 1g.

[0040] The hyperoside content transfer rate of medicinal materials to standard decoction ranges from 35% to 66%.

[0041] Characteristic chromatogram: determined by high performance liquid chromatography (Chinese Pharmacopoeia 2020 Edition, Part IV, General Chapter 0512).

[0042] 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% phosphoric acid solution was used as mobile phase B, with gradient elution performed according to the specifications in the table below; the flow rate was 1.0 mL / min; the column temperature was 30℃; and the detection wavelength was 360 nm. The theoretical plate number, calculated based on the hyperoside peak, should not be less than 5000.

[0043] Table 2 shows the gradient elution table:

[0044]

[0045]

[0046] Preparation of reference solution: Take 1g of Cuscuta chinensis (Southern Cuscuta chinensis) reference material, place it in a stoppered conical flask, add 50ml of 70% methanol, heat under reflux for 30 minutes, cool, shake well, filter, and collect the filtrate as the reference solution. Separately, take appropriate amounts of chlorogenic acid reference standard, hyperoside reference standard, and isoquercitrin reference standard, accurately weigh them, and add methanol to prepare a mixed solution containing 50μg of each per ml, as the reference solution.

[0047] Preparation of test solution: Same as (content determination).

[0048] Assay: Accurately pipette 10 μl of the reference solution and 5 μl of the test solution, inject them into the liquid chromatograph, and determine the result.

[0049] The national standard for "Formula Granules of Salted Cuscuta (Southern Cuscuta)" (No. YBZ-PFKL-2021138) stipulates 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. Among them, 3 peaks should correspond to the retention times of the corresponding reference material peaks. The peak corresponding to the hyperoside reference peak is peak S. The relative retention times of peaks 2 and 3 with peak S are calculated. The relative retention times should be within ±5% of the specified values, which are 0.43 (peak 2) and 0.88 (peak 3). The results show that the characteristic chromatograms of 18 batches of salted dodder seed (southern dodder seed) standard decoction test samples all show chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the reference herb. The RSD of the relative retention times of peaks 1 to 5 is less than 2%, indicating that the elution times of each peak are consistent among batches. Therefore, it is recommended that the relative retention times should be within ±5% of the specified values, which are 0.43 (peak 2) and 0.88 (peak 3).

[0050] The national standard stipulates that the relative peak areas of peaks 3, 5, and S should be within the specified range, which is not less than 0.400 (peak 3) and not less than 0.100 (peak 5). The RSD of the relative peak areas of the characteristic peaks of peaks 3 and 5 in 18 batches of salted dodder seed (southern dodder seed) standard decoction is less than 2%, indicating that the difference in the relative peak areas of peaks 3 and 5 between batches is small. Therefore, it is recommended that the relative peak areas of peaks 3 and 5 and peak 4 (S) be not less than 0.400 (peak 4) and 0.100 (peak 5).

[0051] Example 2:

[0052] 1. Raw material selection

[0053] 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. Salt-cured dodder seed (Southern dodder seed) is a cultivated product, distributed throughout my country. It is mainly produced in Guangdong, Jiangxi, Guangxi, and Fujian provinces, where its cultivation has reached a large scale and is the main source of commercial salt-cured dodder seed (Southern dodder seed). Based on a comprehensive evaluation of raw material quality, sustainable resource supply, and other factors, the preliminary production areas for raw materials used in large-scale production were determined to be Guangdong (Meizhou), Jiangxi (Zhangshu, Wuning, and Ethanol), and Guangxi (Yulin and Guigang). Ultimately, 18 qualified batches were selected for research on standard decoctions.

[0054] 2. Research and standard setting on extract yield and transfer rate

[0055] Table 3 shows the transfer rate of 18 batches of salted dodder seed (southern dodder seed) standard decoction.

[0056]

[0057]

[0058] Based on the content of 18 batches of salted dodder seed (southern dodder seed) slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 50.3%.

[0059] The range calculated using the mean plus or minus 3 times the SD is 41.0% to 59.6%.

[0060] The range calculated using 70% to 130% of the mean is 35.4% to 65.8%.

[0061] Based on the actual range, it is 45.9% to 55.6%;

[0062] It is recommended to use 35% to 66% as the upper and lower limits of the standard decoction transfer rate.

[0063] 3. Content research and standard setting

[0064] Modern research shows that Cuscuta chinensis (Southern Cuscuta) contains various components, such as flavonoids, glycosides, amino acids, and trace elements, as well as cholesterol, brassosterol, sitosterol, stigmasterol, triterpenoid acids, alkaloids, coumarins, tannic acid, and other chemical components. Cuscuta chinensis (Southern Cuscuta) possesses various pharmacological activities, including pregnancy support, aphrodisiac effects, anti-aging properties, and delaying cataract formation. The 2020 edition of the Chinese Pharmacopoeia lists hyperoside as an indicator component for quality control of Cuscuta chinensis (Southern Cuscuta). Therefore, this study selected hyperoside as the quality control indicator component for the standard decoction of Cuscuta chinensis (Southern Cuscuta).

[0065] 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 standard decoction of Cuscuta chinensis (Southern Cuscuta chinensis), the limits and upper and lower limits of glycyrrhizin content in the standard decoction were calculated.

[0066] The range calculated using the mean plus or minus 3 times the SD is 5.3 mg to 8.1 mg.

[0067] The range calculated using 70%–130% of the mean is 4.7 mg–8.7 mg.

[0068] Based on the actual range of 5.8mg to 7.6mg, it is recommended to use the mean value plus or minus 3 times the SD, i.e., 4.7mg to 8.7mg, as the upper and lower limits of the standard decoction content.

[0069] Based on the content of 18 batches of salted dodder seed (southern dodder seed) slices, the content of standard decoction, and the yield of extract, the average transfer rate of standard decoction content was calculated to be 50.3%.

[0070] The range calculated using the mean plus or minus 3 times the SD is 41.0% to 59.6%.

[0071] The range calculated using 70% to 130% of the mean is 35.4% to 65.8%.

[0072] Based on the actual range, it is 45.9% to 55.6%;

[0073] It is recommended to use 35% to 66% as the upper and lower limits of the standard decoction transfer rate.

[0074] 4. Characteristic spectrum research and standard setting

[0075] The standard decoction chromatogram of salted Cuscuta chinensis (Southern Cuscuta chinensis) with experimental batch number BT(YTSZ)211101 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. The generated standard chromatogram is shown below. Figure 3 Hyperoside was selected as the reference peak, and the relative retention times of the characteristic peaks in the chromatogram were calculated.

[0076] By comparing the chromatogram with that of the reference standard, it was found that among the five characteristic peaks in the standard decoction of Cuscuta chinensis (Southern Cuscuta chinensis), peak 4 (S) is hyperoside. According to the "Formula Granules of Cuscuta chinensis (Southern Cuscuta chinensis)" (No. YBZ-PFKL-2021138), peak 1 is chlorogenic acid, peak 2 is cryptochlorogenic acid, and peak 5 is isoquercitrin.

[0077] Table 4 shows the relative retention times of 18 batches of standard decoction of salted dodder seed (southern dodder seed).

[0078]

[0079] Table 5 shows the peak area of ​​18 batches of salted dodder seed (southern dodder seed) standard decoction.

[0080]

[0081]

[0082] The national standard for "Formula Granules of Salted Cuscuta (Southern Cuscuta)" (No. YBZ-PFKL-2021138) stipulates 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. Among them, 3 peaks should correspond to the retention times of the corresponding reference material peaks. The peak corresponding to the hyperoside reference peak is peak S. The relative retention times of peaks 2 and 3 with peak S are calculated. The relative retention times should be within ±10% of the specified values, which are 0.43 (peak 2) and 0.88 (peak 3). The results show that the characteristic chromatograms of 18 batches of salted dodder seed (southern dodder seed) standard decoction test samples all show chromatographic peaks corresponding to the retention times of the five main characteristic peaks in the reference herb. The RSD of the relative retention times of peaks 1 to 5 is less than 2%, indicating that the elution times of each peak are consistent among batches. Therefore, it is recommended that the relative retention times be within ±5% of the specified values, which are 0.43 (peak 2) and 0.88 (peak 3).

[0083] The national standard stipulates that the relative peak areas of peaks 3, 5, and S should be within the specified range, which is not less than 0.400 (peak 3) and not less than 0.100 (peak 5). The RSD of the relative peak areas of the characteristic peaks of peaks 3 and 5 in 18 batches of salted dodder seed (southern dodder seed) standard decoction is less than 2%, indicating that the difference in the relative peak areas of peaks 3 and 5 between batches is small. Therefore, it is recommended that the relative peak areas of peaks 3 and 5 and peak 4 (S) be not less than 0.400 (peak 4) and 0.100 (peak 5).

[0084] 5. Leachate research and standard setting

[0085] 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 under the "Determination of Alcohol-Soluble Extracts" section of the Chinese Pharmacopoeia 2020 General Chapter 2201. The test results of 18 batches of standard decoction of salted dodder seed (Southern dodder seed) are shown in Table 6.

[0086] Table 6 shows the results of extract determination for 18 batches of salted dodder (southern dodder).

[0087]

[0088]

[0089] The average value of the alcohol-soluble extract from 18 batches of standard decoctions was 25.6%.

[0090] The range calculated using the mean plus or minus 3 times the SD is 10.6% to 40.6%.

[0091] The range calculated using 70% to 130% of the mean is 17.9% to 33.3%.

[0092] Based on the actual range, it is 19.5% to 34.7%;

[0093] It is recommended to take 17.9% to 40.6% as the upper and lower limits of the standard decoction extract.

[0094] 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.

[0095] 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 salted dodder seed, firstly, weigh the salted dodder seed (southern dodder 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 salted dodder 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 as salted dodder seed slices 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 decoction of salted dodder seed according to claim 1, characterized in that, In Step 4, vacuum concentration is carried out under an absolute vacuum of 30–40 mbar.

Images