A kind of clear-fried licorice formula granule and its preparation method and quality control method

Abstract

This application provides a stir-fried licorice formula granule, its preparation method, and quality control method. The method comprises: the stir-fried licorice formula granule is prepared from stir-fried licorice slices as raw material through water extraction, vacuum concentration, drying, and granulation. Each gram of formula granule is equivalent to 1.7-2.1 grams of stir-fried licorice slices, wherein the glycyrrhizin content is not less than 1.0%, and the glycyrrhizic acid content is not less than 2.1%. This invention also establishes a comprehensive quality control method for the formula granules. High-performance liquid chromatography (HPLC) is used to determine the content of key components, combined with chemical fingerprinting to evaluate overall chemical consistency, and physical fingerprinting is introduced to monitor the physical stability of the granules, forming a three-in-one quality control system of "component-chemistry-physics". This invention fills the technological gap in stir-fried licorice formula granules and provides a complete solution for their standardized production and safe and effective application.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine and natural medicine pharmaceutical manufacturing, and in particular to a stir-fried licorice formula granule, its preparation method, and quality control method. Background Technology

[0002] Licorice is the dried root and rhizome of *Glycyrrhiza uralensis*, *Glycyrrhiza inflata*, or *Glycyrrhiza glabra*, belonging to the legume family. It was first recorded in the *Shennong Bencao Jing* (Shennong's Classic of Materia Medica) and listed as a superior-grade herb. The Chinese Pharmacopoeia records that raw licorice is sweet and neutral in nature, possessing the functions of tonifying the spleen and replenishing qi, clearing heat and detoxifying, resolving phlegm and relieving cough, alleviating spasms and pain, and harmonizing other herbs. Ancient classic prescriptions and compound preparations of traditional Chinese medicine have been included in the current registration classification of Chinese medicine. In the three batches of the *Ancient Classic Prescriptions Catalogue* published by relevant units, totaling 331 prescriptions, 107 prescriptions contain licorice, accounting for 32.3%, of which 40 are labeled as "processed" or "processed licorice," accounting for 37.4%. In clinical practice, licorice often needs to be processed to alter or enhance certain medicinal effects. Stir-frying is one of its important processing methods. By stir-frying over a low flame, its cooling properties are reduced, its drying properties are enhanced, making it more effective in tonifying the spleen and replenishing qi, protecting the stomach qi, and effectively moderating its medicinal properties to prevent the adverse effects of overdose on abdominal distension (fullness in the middle jiao). According to ancient books such as the *Jin Gui Yu Han Jing*, the "roasted licorice" recorded in prescriptions from the Han Dynasty and earlier was actually prepared using the stir-frying method. Therefore, when formulating the technical documents accompanying the *Catalogue of Famous Ancient Prescriptions*, relevant national departments explicitly designated the "roasted licorice" used in classic prescriptions such as Houpu Wenzhong Tang and Danggui Sini Tang as stir-fried licorice.

[0003] With the modernization of traditional Chinese medicine (TCM), TCM granules have become an important supplement to traditional Chinese medicine decoction pieces due to their advantages such as convenient portability, accurate dosage, and controllable quality, meeting the clinical needs of modern fast-paced life. The National Pharmacopoeia Commission has successively promulgated hundreds of national standards for TCM granules, and the industry is in a stage of rapid development towards standardization and regulation. However, existing national standards and mainstream research focus on raw licorice granules and honey-processed licorice granules; the preparation process and quality standards for stir-fried licorice granules remain lacking, with a lack of unified national standards and mature industrialization technologies. Stir-fried licorice and its granules are in high demand both in classic prescriptions and in clinical dispensing. Therefore, effectively standardizing the preparation process of stir-fried licorice granules and establishing appropriate quality control methods are of great significance for ensuring the stable and reliable quality of stir-fried licorice granules and their safe and effective clinical application. Summary of the Invention

[0004] To address the lack of stir-fried licorice formula granules in the development and clinical dispensing of classic prescriptions, and the absence of standardized preparation processes and systematic quality control methods in existing technologies, this invention provides a stable and controllable stir-fried licorice formula granule, and establishes a complete preparation method and comprehensive quality control system for it.

[0005] The first aspect of the present invention provides a stir-fried licorice formula granule, comprising: a solid preparation made from stir-fried licorice slices as raw material by water extraction, vacuum concentration, drying and granulation processes; wherein the stir-fried licorice formula granule contains glycyrrhizin, glycyrrhizic acid and glycyrrhizin, and the content of glycyrrhizin is not less than 1.0% and the content of glycyrrhizic acid is not less than 2.1%.

[0006] Furthermore, each gram of stir-fried licorice granules is equivalent to 1.7~2.1g of stir-fried licorice slices.

[0007] A second aspect of this invention provides a method for preparing stir-fried licorice root granules, comprising the following steps: S1. Extraction: Weigh the stir-fried licorice slices, add water and heat under reflux to extract, filter and collect the filtrate; S2. Concentration: The filtrate obtained in step S1 is concentrated under reduced pressure at 60~70℃ to obtain a clear extract; S3. Drying and pulverizing: The clear extract obtained in step S2 is dried under reduced pressure at 70~80℃ to obtain a dry extract, and then pulverized to obtain a dry extract powder; S4. Granulation: The dry powder obtained in step S3 is mixed with excipients, and then granulated after being made into a soft mass with an 80%~90% ethanol solution. The granules are then dried at 55℃~65℃ to obtain stir-fried licorice formula granules.

[0008] Furthermore, the heating and reflux extraction in step S1 is performed 2 to 3 times, with each extraction lasting 1.5 to 2.5 hours, and the amount of water added is 8 to 12 times the weight of the medicinal slices.

[0009] Furthermore, the excipient mentioned in step S4 is dextrin, and the mass ratio of the dry extract powder to dextrin is 1.5-2.5:1.

[0010] A third aspect of this invention provides a quality control method for stir-fried licorice root granules, comprising the following steps: SS1. Determination of active ingredient content: The contents of glycyrrhizin and glycyrrhizic acid in the formula granules were simultaneously determined by high performance liquid chromatography. SS2, Chemical fingerprint evaluation: Establish the high performance liquid chromatography fingerprint of the formulated particles and calculate the similarity between the fingerprint and the control fingerprint. SS3. Physical fingerprint spectrum evaluation: Multiple physical indicators of the formulated particles are measured, and after standardization, a physical fingerprint spectrum is generated and a similarity evaluation is performed.

[0011] Furthermore, in steps SS1 and SS2, the chromatographic conditions for the high-performance liquid chromatography include: Waters Symmetry C18 columns were used, with dimensions of 4.6mm × 250mm and a thickness of 5μm. Mobile phase A is acetonitrile, and mobile phase B is a 0.05% aqueous solution of phosphoric acid; The gradient elution program was set as follows: 0–15 min, phase A ratio 17%–19%; 15–30 min, phase A ratio 19%–30%; 30–55 min, phase A ratio 30%–50%. The flow rate was 1.0 mL / min, the detection wavelength was 237 nm, the column temperature was 25 °C, and the injection volume was 10 μL.

[0012] Furthermore, in step SS2, the similarity is not less than 0.90.

[0013] Furthermore, in step SS3, the multiple physical indicators include moisture content, moisture absorption rate, angle of repose, Hausner ratio, bulk density, tap density, and relative uniformity index.

[0014] Compared with the prior art, the present invention has the following beneficial effects: 1) For the first time, a complete production technology solution for stir-fried licorice formula granules was constructed, filling the technological gap in the development and clinical application of classic prescriptions for stir-fried licorice formula granules.

[0015] 2) By clarifying the core parameters of key processes such as water extraction, vacuum concentration / drying, and granulation, a stable and repeatable standardized preparation process was established, ensuring the quality consistency of the conversion process from medicinal slices to granules, and providing a reliable basis for large-scale production.

[0016] 3) A comprehensive quality control system integrating "content determination, chemical fingerprinting, and physical fingerprinting" has been established.

[0017] 4) The quantitative conversion relationship between decoction pieces and granules was clarified (1g granules≈1.7~2.1g decoction pieces), ensuring the accuracy of clinical dosage and medication safety.

[0018] 5) A scalable quality control paradigm has been established, providing a systematic solution for the in-depth development of similar Chinese herbal medicine formula granules. Attached Figure Description

[0019] Figure 1 This is a typical liquid chromatogram of stir-fried licorice granules.

[0020] Figure 2 This is the liquid chromatogram of a mixed reference standard of glycyrrhizin and glycyrrhizic acid.

[0021] Figure 3The fingerprint chromatograms are of 15 batches of stir-fried licorice formula granules; in the figure, K1-K15 represent the fingerprint chromatograms of 15 batches of stir-fried licorice formula granules; R(10) is the generated control fingerprint chromatogram; peaks 1-10 are 10 common peaks; peak 3 is glycyrrhizin; peak 5 is glycyrrhizin; peak 9 is glycyrrhizic acid.

[0022] Figure 4 The figures show the physical fingerprint spectra of 15 batches of stir-fried licorice formula granules and the control physical fingerprint spectra; Figure A shows the physical fingerprint spectra of 15 batches of stir-fried licorice formula granules (K1-K15); Figure B shows the generated control physical fingerprint spectra. Detailed Implementation

[0023] To enable those skilled in the art to better understand the technical solutions in this specification, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this specification, and not all embodiments. Based on the embodiments in this specification, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of this specification.

[0024] Test instruments and materials The instruments and materials used in the specific implementation of this invention are as follows: Experimental instruments: Agilent 1260 high performance liquid chromatograph (Agilent Technologies); GZX-9146MBE digital display drying oven (Shanghai Boxun Medical Biological Instrument Co., Ltd.); BSA1248-CW electronic balance (Beijing Sartorius Scientific Instruments Co., Ltd.); XS3DU balance (Mettler Toledo International Ltd.); ultrasonic cleaner (Dongguan Jiekang Ultrasonic Equipment Co., Ltd.); N-1300 rotary evaporator (EYELA Tokyo Rika); MZ-0.3M3 pulsed vacuum dryer (Changzhou Zhenhua Drying Equipment Co., Ltd.).

[0025] Raw materials and reagents: 15 batches of licorice slices from different origins (details in Table 1); glycyrrhizin (batch number 111610-202209, purity 95.2%, purchased from the National Institutes for Food and Drug Control); ammonium glycyrrhizate reference standard (batch number 110731-202122, purity 94.4%, purchased from the National Institutes for Food and Drug Control); acetonitrile (chromatographic grade, Merck AG, Germany); phosphoric acid (chromatographic grade, Tianjin Kemeio Chemical Reagent Co., Ltd.); Wahaha purified water; other reagents were all analytical grade.

[0026] Example 1 Preparation of stir-fried licorice slices: Take 15 batches of licorice slices from Table 1 (raw material source and batch number are shown in Table 1) and refer to the processing method under "Stir-fried Licorice" in the "Standards for Processing Traditional Chinese Medicine Slices in Zhejiang Province" (Zhejiang Provincial Food and Drug Administration. Standards for Processing Traditional Chinese Medicine in Zhejiang Province [M]. Beijing: China Medical Science and Technology Press, 2015: 23.): "Take licorice slices and stir-fry them according to the stir-frying method until the surface is deep yellow and slightly charred. Then take them out and let them cool."

[0027] The specific preparation process is as follows: 100g of licorice slices (numbered S1~S15) are placed in a preheated stir-frying machine, the temperature is controlled at 120℃, the stir-frying time is 25min, and when the surface is dark yellow and slightly charred, they are taken out and cooled to obtain 15 batches of stir-fried licorice slices, which are numbered P1~P15 in sequence. The quality of all batches meets the above-mentioned "Zhejiang Province Traditional Chinese Medicine Slices Processing Specifications".

[0028] Table 1. Sources of raw licorice slices and the serial numbers of 15 batches of stir-fried licorice slices obtained. Example 2 Preparation of stir-fried licorice granules: This invention uses the aforementioned 15 batches of stir-fried licorice slices as raw materials to prepare 15 batches of stir-fried licorice formula granules. The preparation process for each batch is as follows: Take 100g of stir-fried licorice slices, add 10 times the amount of water and soak for 30 minutes, then heat and reflux for 2 hours, and filter while hot. Add 10 times the amount of water to the residue and heat and reflux twice more, each time for 2 hours. Filter while hot, combine the filtrates, and concentrate the filtrate under reduced pressure at 65℃ to obtain a clear extract. Place the clear extract in a pulsed vacuum dryer and dry under reduced pressure at 75℃ for 9 hours until it reaches a dry extract state, then pulverize it into a fine powder. Add an appropriate amount of dextrin (the ratio of dry extract powder to dextrin is 2:1) to the dry extract powder, mix well, prepare a soft mass with an appropriate amount of 85% ethanol, granulate, and dry at 60℃ for 4 hours to obtain stir-fried licorice formula granules. Using 15 batches of stir-fried licorice slices (P1~P15) in Table 1, 15 batches of stir-fried licorice formula granules (corresponding numbers K1~K15) were prepared, as shown in Table 2.

[0029] Table 2. Fifteen batches (K1~K15) of stir-fried licorice granules prepared. Example 3 Determination of the content (glycyrrhizin and glycyrrhizic acid) in stir-fried licorice powder: 1. Chromatographic conditions: The chromatographic column was a Waterssymmetry C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile (A) - 0.05% phosphoric acid solution (B), with gradient elution (0–15 min, 17–19% A; 15–30 min, 19%–30% A; 30–55 min, 30%–50% A); the flow rate was 1 mL / min; the detection wavelength was 237 nm; the column temperature was 25 °C; and the injection volume was 10 μL.

[0030] 2. Preparation of reference solution: Take appropriate amounts of glycyrrhizin reference standard and ammonium glycyrrhizate reference standard, accurately weigh them, and add 70% ethanol to prepare 1 ml solutions containing 20 μg of glycyrrhizin and 0.2 mg of ammonium glycyrrhizate, respectively, to obtain the reference solution (weight of glycyrrhizic acid = weight of ammonium glycyrrhizate / 1.0207).

[0031] 3. Preparation of the test solution: Accurately weigh 0.2g of stir-fried licorice granules and place them in a 100mL stoppered conical flask. Accurately add 25mL of 70% ethanol, weigh the flask, and sonicate (500W power, 40kHz frequency) for 30min. Cool the flask, weigh it again, and replenish the lost mass with 70% ethanol. Shake well. Filter the flask and collect the filtrate to obtain the test solution.

[0032] 4. Methodological validation: Method validation was performed on the content determination method, including linearity, precision, stability, repeatability, and recovery tests. Specific test procedures are as follows: ① Linear relationship experiment: Regression was performed with the concentration (mg / ml) of each reference standard as the abscissa (x) and the peak area as the ordinate (y) (see Table 3). The regression equations for glycyrrhizin and glycyrrhizic acid were y=10865x+13.617 (R²=0.9996) and y=5487.8x+4.3774 (R²=1), respectively. This indicates that glycyrrhizin has a good linear relationship in the concentration range of 0.0030–0.1933 mg / ml, and glycyrrhizic acid has a good linear relationship in the concentration range of 0.0108–0.6930 mg / ml.

[0033] Table 3. Results of linear relationship test for determination of glycyrrhizin and glycyrrhizic acid content. ②Precision test: The solution was prepared according to the above method, and the injection was repeated 6 times under the above chromatographic conditions. The peak area was measured and the RSD value was calculated (see Table 4). The RSD values ​​of glycyrrhizin and glycyrrhizic acid were 0.09% and 0.81% respectively (n=6), indicating that the instrument has good precision.

[0034] Table 4. Precision test results of the method for determining glycyrrhizin and glycyrrhizic acid content. ③Stability test: The solution was prepared according to the above method, and injected at 0, 2, 4, 6, 8, 10, 12, and 24 h under the above chromatographic conditions. The peak area was measured and the RSD value of the peak area was calculated (see Table 5). The RSD values ​​of glycyrrhizin and ammonium glycyrrhizate were 0.21% and 0.64% (n=6), respectively, indicating that the solution had good stability within 24 h.

[0035] Table 5. Stability test results of the method for determining glycyrrhizin and glycyrrhizic acid content. ④ Repeatability test: Six solutions were prepared according to the above-described method for preparing the test solution. The peak areas were measured and the contents of glycyrrhizin and glycyrrhizic acid were calculated under the above chromatographic conditions (see Table 6). The results showed that the average contents of glycyrrhizin and glycyrrhizic acid in the repeatability test were 1.33% and 2.38%, respectively, and the RSDs of glycyrrhizin and glycyrrhizic acid were 0.93% and 1.87% (n=6), respectively, indicating that the method has good repeatability.

[0036] Table 6. Results of repeatability tests for the determination methods of glycyrrhizin and glycyrrhizic acid content. ⑤ Recovery test: Appropriate amounts of glycyrrhizin and glycyrrhizic acid ammonium reference standards were added to six samples of stir-fried licorice root with known concentrations. The peak areas were determined and RSD values ​​were calculated under the chromatographic conditions described above (see Tables 7 and 8). The results showed that the average recoveries of glycyrrhizin and glycyrrhizic acid ammonium were 101.18% and 100.40%, respectively, with RSDs of 0.91% and 2.33% (n=6), indicating good accuracy of the above chromatographic conditions.

[0037] Table 7 Results of glycyrrhizin recovery test Table 8 Results of glycyrrhizic acid recovery test 5. Results of glycyrrhizin and glycyrrhizic acid content determination: Fifteen batches of stir-fried licorice granules (K1~K15) were taken, and solutions were prepared according to the above-described method for preparing test solutions. The solutions were then injected and analyzed under the above-described chromatographic conditions. The contents of glycyrrhizin and glycyrrhizic acid in the samples were calculated using the standard curve method. The results are shown in Table 9. The content range of glycyrrhizin was 1.03%~2.66%, and the content range of glycyrrhizic acid was 2.19%~3.91%. A typical liquid chromatogram for the content determination of stir-fried licorice granules is shown in Table 9. Figure 1 ), liquid chromatogram of glycyrrhizin and glycyrrhizic acid mixed reference standard (see Figure 2 ).

[0038] Table 9. Results of determination of glycyrrhizin and glycyrrhizic acid content in 15 batches of stir-fried licorice granules. Example 4 Establishment and evaluation of high-performance liquid chromatography fingerprints: 1. Chromatographic conditions: The chromatographic column was a Waterssymmetry C18 (4.6 mm × 250 mm, 5 μm); the mobile phase was acetonitrile (A) - 0.05% phosphoric acid solution (B), with gradient elution (0–15 min, 17–19% A; 15–30 min, 19%–30% A; 30–55 min, 30%–50% A); the flow rate was 1 mL / min; the detection wavelength was 237 nm; the column temperature was 25 °C; and the injection volume was 10 μL.

[0039] 2. Preparation of reference solution: Take appropriate amounts of glycyrrhizin reference standard and ammonium glycyrrhizate reference standard, accurately weigh them, and add 70% ethanol to prepare 1 ml solutions containing 20 μg of glycyrrhizin and 0.2 mg of ammonium glycyrrhizate, respectively, to obtain the reference solution (weight of glycyrrhizic acid = weight of ammonium glycyrrhizate / 1.0207).

[0040] 3. Preparation of the test solution: Accurately weigh 0.2g of stir-fried licorice granules and place them in a 100mL stoppered conical flask. Accurately add 25mL of 70% ethanol, weigh the flask, and sonicate (500W power, 40kHz frequency) for 30min. Cool the flask, weigh it again, and replenish the lost mass with 70% ethanol. Shake well. Filter the flask and collect the filtrate to obtain the test solution.

[0041] 4. Methodological validation: Methodological validation was performed on the preparation method of the test solution, including precision, repeatability, and stability tests. Specific test contents are as follows: ①Precision test: Take the stir-fried licorice granules (No. K10), prepare a solution according to the above-described method for preparing the test solution, and continuously inject and determine six times under the above chromatographic conditions, recording the peak area. After identification with the reference standard, peak 3 was identified as the glycyrrhizin chromatographic peak. The retention times in the six consecutive injections were 16.3470, 16.6880, 16.7040, 16.6410, 16.6240, and 16.6760 min, respectively, and the peak areas were 1165.4, 1167.8, 1165.1, 1166.8, 1166.6, and 1165.8 min, respectively. Using peak 3 (S) as the glycyrrhizin chromatographic peak as the reference peak, the relative retention times (Table 10) and relative peak areas (Table 11) of each of the 10 common peaks were calculated. The RSDs obtained were all less than 1.42%, indicating that the method has good precision.

[0042] Table 10 Precision test results (relative retention time) Table 11 Precision test results (relative peak area) ② Repeatability test: Six parallel test solutions were prepared using stir-fried licorice granules (K10) according to the above-described test solution preparation method. These solutions were then continuously injected and analyzed under the aforementioned chromatographic conditions, and the peak areas were recorded. After identification with the reference standard, peak 3 was confirmed to be the glycyrrhizin chromatographic peak. The retention times in the six consecutive injections were 16.7040, 16.4760, 16.4880, 16.4090, 16.4350, and 16.1410 min, respectively, and the peak areas were 1165.1, 1210.5, 1183.2, 1175.90, 1180.0, and 1173.5, respectively. Using peak 3 (the glycyrrhizin chromatographic peak) as the reference peak, the relative retention times (Table 12) and relative peak areas (Table 13) of the ten common peaks were calculated. The RSDs were all less than 2.64%, indicating good repeatability of the method.

[0043] Table 12 Results of repeatability tests (relative retention time) Table 13 Results of repeatability tests (relative peak area) ③Stability test: Take stir-fried licorice granules (K10) and prepare a solution according to the above-described method for preparing the test solution. Inject and determine the peak area at 0, 2, 4, 8, 10, 12, and 24 hours under the above chromatographic conditions. Peak 3 was identified as glycyrrhizin by the reference standard. Its retention times in six consecutive injections were 16.3470, 16.7040, 16.6240, 16.5650, 16.4770, and 16.1290 min, respectively, and its peak areas were 1165.4, 1165.1, 1166.6, 1167.2, 1166.5, and 1160.5 min, respectively. Using peak 3 (glycyrrhizin peak) as the reference peak, calculate the relative retention times (Table 14) and relative peak areas (Table 15) of each of the ten common peaks. The RSDs were all less than 2.77%, indicating that the test solution has good stability within 24 hours at room temperature.

[0044] Table 14 Stability test results (relative retention time) Table 15 Stability test results (relative peak area) 5. Establishment of fingerprint pattern: Fifteen batches of stir-fried licorice root granules were prepared in parallel, and the solutions were injected and analyzed under the chromatographic conditions described above. The chromatograms were recorded. The HPLC chromatograms of the 15 batches of stir-fried licorice root granules were imported into the 2012 version of the "Evaluation Software for Chromatographic Fingerprints of Traditional Chinese Medicines". Using the K10 fingerprint chromatogram as a reference chromatogram, with a time window width of 0.1 min, the median method was used. After multi-point correction and Mark peak matching, the fingerprint chromatograms of the 15 batches of stir-fried licorice root granules and the reference fingerprint chromatogram R were generated (see [link to software]). Figure 3 The HPLC chromatogram of the mixture was compared with that of the mixed reference standard, and the components corresponding to peaks 3, 5 and 9 were glycyrrhizin, glycyrrhizin and glycyrrhizic acid, respectively.

[0045] 6. Fingerprint similarity calculation: Using the 2012 edition of the "Chromatographic Fingerprint Evaluation Software for Traditional Chinese Medicine" provided by the National Pharmacopoeia Commission, the chromatograms of 15 batches of stir-fried licorice root granules (K1~K15) were sequentially imported into the software, and the similarity of the 15 batches of stir-fried licorice root granules was calculated (see Table 16). The similarity of the 15 batches of stir-fried licorice root granules was not less than 0.968, indicating that the types of chemical components contained in the 15 batches of stir-fried licorice root granules were basically the same.

[0046] Table 16. Similarity Evaluation Results of 15 Batches of Stir-fried Licorice Granules Example 5 Establishment and evaluation of the physical fingerprint spectrum of stir-fried licorice granules: 1. Indicator Measurement: Based on the physical properties of stir-fried licorice root granules, flowability, stability, bulking capacity, and uniformity were selected as primary indicators; moisture content, hygroscopicity, angle of repose, Hausner ratio, loose density, tap density, and relative homogeneity index were selected as secondary indicators (characterizing the physical properties of the stir-fried licorice root granule samples). The batches of stir-fried licorice root granules to be tested were analyzed to obtain the measured data of their secondary indicators. The methods for determining the secondary indicators are as follows: ① Moisture content: The moisture content of 15 batches of stir-fried licorice formula granules was determined according to Method II of General Chapter 0832 in the 2025 edition of the Chinese Pharmacopoeia.

[0047] ② Moisture absorption rate: Equilibrate the solution containing saturated sodium chloride in a desiccator for 24 hours, then take 2g of granules and place them in a constant-weight volumetric flask, place it in the above-mentioned equilibrated desiccator, and place it under constant temperature and humidity (75% relative humidity, 25℃) for 24 hours. Take it out, weigh it, and calculate the moisture absorption rate.

[0048] ③ Angle of repose: Connect two identical funnels in series and fix them on an iron stand at a certain height H above the paper. Slowly pour the dried granules along the funnel wall until the tip of the small cone formed by the extract powder on the paper touches the funnel opening. Read the diameter 2R of the cone on the graph paper and calculate the angle of repose. The formula is angle of repose = acrtan(H / R). Calculate the average value of the two parallel calculations.

[0049] ④ Loose density: Take a dry, clean 10ml graduated cylinder, accurately weigh a certain mass (M) of particles, slowly add them into the cylinder, and read the volume V. a To calculate bulk density, use the formula: Bulk density = M / V a .

[0050] ⑤ Tapped density: Vibrate the graduated cylinder containing the particles in ④ up and down 200 times at a frequency of 2 seconds per cycle, and read the volume V. c Calculate the tap density using the formula: Tap density = M / V c .

[0051] ⑥ Hausnerby: The formula is Hausnerby = tapped density / loose density.

[0052] ⑦ Relative homogeneity index: Pass the particles sequentially through sieves No. 1 to No. 9, vibrate for 5 minutes, and record the mass of particles retained by each sieve. Take particles retained by sieves with average apertures of 2000, 1425, 602.5, 302.5, 215, 165, 137.7, 107.5, and 82.5 μm, respectively, and calculate the relative homogeneity index. The formula is: Relative homogeneity index = F m / [100+(d m -d m-1)×F m-1 +(d m+1 -d m )×F m+1 +(d m -d m-2 )×F m-2 +...+(d m +nd m )×F m+n ], where F m F represents the mass percentage of particle size over most ranges. m+1 F represents the percentage of particles retained by a single sieve across most particle size ranges. m-1 d represents the percentage of particles retained by the next sieve layer across most particle size ranges. m d represents the average particle size across most particle size ranges. m+1 d represents the average particle size retained by one sieve layer across most particle size ranges. m-1 The average particle size retained by the next layer of sieves in most particle size ranges, where n is the number of defined particle size ranges.

[0053] 2. Standardized transformation: To eliminate the influence of dimensions, the physical indicators are standardized, i.e., converted to the same range of 0 to 10. The conversion formula is shown in Table 17, and the conversion results are shown in Table 18.

[0054] Table 17 Standardized Conversion Formulas for Physical Indicators Table 18 Results of Standardization Conversion of Physical Indicators 3. Atlas generation and similarity analysis: The detection data of the secondary indicators are standardized and used to construct the physical fingerprint spectrum of the stir-fried licorice formula granules.

[0055] The standardized values ​​of the secondary physical indicators of 15 batches of samples were used to create physical fingerprint spectra in an Excel spreadsheet. Then, radar charts were plotted using the average of the standardized values ​​to establish a control physical fingerprint spectrum. The results are shown below. Figure 4 The similarity results of samples K1 to K15 were compared using SPSS 27.0 software and similarity analysis was performed using the Pearson correlation coefficient method. The similarity results were 0.987, 0.990, 0.997, 0.991, 0.988, 0.992, 0.985, 0.983, 0.998, 0.982, 0.984, 0.986, 0.996, 0.977, and 0.975, respectively, all greater than 0.975, indicating that the samples from different batches have good stability and consistency.

[0056] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any changes or substitutions conceived without inventive effort should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope defined in the claims.

[0057] The above-described embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A granule formula for stir-fried licorice root, characterized in that, A solid preparation made from stir-fried licorice slices as raw material through water extraction, vacuum concentration, drying and granulation processes; wherein the stir-fried licorice formula granules contain glycyrrhizin, glycyrrhizic acid and glycyrrhizin, and the content of glycyrrhizin is not less than 1.0% and the content of glycyrrhizic acid is not less than 2.1%.

2. The stir-fried licorice granules according to claim 1, characterized in that, Each gram of stir-fried licorice granules is equivalent to 1.7~2.1g of stir-fried licorice slices.

3. The method for preparing the stir-fried licorice formula granules according to claim 1 or 2, characterized in that, Includes the following steps: S1. Extraction: Weigh the stir-fried licorice slices, add water and heat under reflux to extract, filter and collect the filtrate; S2. Concentration: The filtrate obtained in step S1 is concentrated under reduced pressure at 60~70℃ to obtain a clear extract; S3. Drying and pulverizing: The clear extract obtained in step S2 is dried under reduced pressure at 70~80℃ to obtain a dry extract, and then pulverized to obtain a dry extract powder; S4. Granulation: The dry powder obtained in step S3 is mixed with excipients, and then granulated after being made into a soft mass with an 80%~90% ethanol solution. The granules are then dried at 55℃~65℃ to obtain stir-fried licorice formula granules.

4. The preparation method according to claim 3, characterized in that, The heating and reflux extraction in step S1 is performed 2 to 3 times, with each extraction lasting 1.5 to 2.5 hours, and the amount of water added is 8 to 12 times the weight of the medicinal slices.

5. The preparation method according to claim 3, characterized in that, The excipient mentioned in step S4 is dextrin, and the mass ratio of the dry extract powder to dextrin is 1.5-2.5:

1.

6. The quality control method for stir-fried licorice root granules according to claim 1 or 2, characterized in that, Includes the following steps: SS1. Determination of active ingredient content: The contents of glycyrrhizin and glycyrrhizic acid in the formula granules were simultaneously determined by high performance liquid chromatography. SS2, Chemical fingerprint evaluation: Establish the high performance liquid chromatography fingerprint of the formulated particles and calculate the similarity between the fingerprint and the control fingerprint. SS3. Physical fingerprint spectrum evaluation: Multiple physical indicators of the formulated particles are measured, and after standardization, a physical fingerprint spectrum is generated and a similarity evaluation is performed.

7. The quality control method according to claim 6, characterized in that, In steps SS1 and SS2, the chromatographic conditions for the high-performance liquid chromatography include: Waters Symmetry C18 columns were used, with dimensions of 4.6mm × 250mm and a thickness of 5μm. Mobile phase A is acetonitrile, and mobile phase B is a 0.05% aqueous solution of phosphoric acid; The gradient elution program was set as follows: 0–15 min, phase A ratio 17%–19%; 15–30 min, phase A ratio 19%–30%; 30–55 min, phase A ratio 30%–50%. The flow rate was 1.0 mL / min, the detection wavelength was 237 nm, the column temperature was 25 °C, and the injection volume was 10 μL.

8. The quality control method according to claim 6, characterized in that, In step SS2, the similarity is not less than 0.

90.

9. The quality control method according to claim 6, characterized in that, In step SS3, the multiple physical indicators include moisture content, moisture absorption rate, angle of repose, Hausner ratio, bulk density, tap density, and relative uniformity index.

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