Method for establishing HPLC fingerprint spectrum of Tongmai granules and fingerprint spectrum thereof

Abstract

The invention provides a Tongmai particle HPLC fingerprint establishment method and a fingerprint thereof, and belongs to the technical field of fingerprints, and the method comprises the following steps: preparing a test solution, taking 0.1-0.5 g of Tongmai particle fine powder, precisely weighing, cooling, complementing the reduced weight, and filtering with a 0.45 [mu] m filter membrane to obtain the test solution, comprising the following raw materials: 500 parts of radix salviae miltiorrhizae, 500 parts of rhizoma chuanxiong and 500 parts of radix puerariae. Taking puerarin and salvianolic acid B reference substances, and setting a gradient elution program; performing similarity evaluation by adopting a traditional Chinese medicine chromatographic fingerprint similarity evaluation system; through high performance liquid chromatography-fingerprint analysis, synchronous quantification of multiple components is realized, an HPLC fingerprint method covering water-soluble components at the same time is established, synchronous monitoring of multiple characteristic peaks of Tongmai particles is realized, the limitation of traditional single component detection is solved, and the stability of a production process is objectively quantified through similarity evaluation.

Description

Technical Field

[0001] The invention belongs to the technical field of fingerprints, in particular to a method for establishing a HPLC fingerprint of Tongmai granules and the fingerprint thereof. Background Art

[0002] Tongmai Granules is a classic small compound prescription for treating cardiocerebral ischemia. It is composed of only three classic Chinese medicines for treating cardiocerebral ischemia. The daily dosage of the raw medicine is 45g. Combined with the pharmacological activity analysis of the Chinese medicine in the prescription, Tongmai Granules has a good effect in treating cardiocerebral ischemia, arteriosclerosis, cerebral thrombosis, cerebral ischemia, coronary heart disease, angina pectoris and other aspects.

[0003] In existing technologies, the transfer rate of active ingredients is unstable due to process differences. Traditional testing only targets a single ingredient and cannot fully reflect product quality. In addition, there are many ingredients in the product, and it is impossible to confirm the method one by one and conduct quantitative analysis in a short period of time.

[0004] In view of the above problems, the present invention proposes a method for establishing an HPLC fingerprint of Tongmai Granules and its fingerprint. Through high-performance liquid chromatography-fingerprint analysis, simultaneous quantification of multiple components is achieved, and an HPLC fingerprint method that simultaneously covers water-soluble components is established to achieve simultaneous monitoring of multiple characteristic peaks of Tongmai Granules, thereby solving the limitations of traditional single component detection and objectively quantifying the stability of the production process through similarity evaluation. Summary of the Invention

[0005] The purpose of the present invention is to address the above-mentioned problems in the existing technology and propose a method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint.

[0006] The purpose of the present invention can be achieved through the following technical solutions:

[0007] The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof comprise the following steps:

[0008] S1: Prepare the test solution. Take 0.1-0.5g of Tongmai granule powder, accurately weigh it, add 25ml of 30% ethanol or 70% methanol solution, and ultrasonically treat it for 30min at a power of 300W and a frequency of 40kHz. After cooling, make up the weight loss, and filter it through a 0.45μm filter membrane to obtain the test solution.

[0009] Among them, Tongmai Granules include the following ingredients in terms of parts: 500 parts of Danshen, 500 parts of Chuanxiong, and 500 parts of Pueraria;

[0010] S2: Take puerarin and salvianolic acid B reference substances and add solvent to make a solution containing 0.5 mg per 1 ml;

[0011] S3: Select chromatographic conditions:

[0012] S31: Chromatographic column: octadecylsilane bonded silica gel column (Agilent ZORBAX SB-C18, 4.6 mm × 250 mm, 5 μm);

[0013] S32: mobile phase A: acetonitrile-water solution (10:90), mobile phase B: acetonitrile-0.1% phosphoric acid solution (22:78);

[0014] S33: set gradient elution program;

[0015] S34: Set parameters: detection wavelength: 250 nm or 286 nm, flow rate: 1.0 mL / min, column temperature: 30°C, injection volume: 10 μL;

[0016] S3: Inject the test solution and reference solution separately and record the chromatogram;

[0017] S4: Similarity evaluation is performed using the "Chinese Herbal Medicine Chromatographic Fingerprint Similarity Evaluation System", and the similarity must not be less than 0.90;

[0018] S5: Test the Tongmai Granules to be tested, evaluate the results by the similarity of the fingerprint patterns, and determine whether the quality meets the requirements according to the quality standards.

[0019] Preferably, the preparation of the Tongmai granules comprises the following steps:

[0020] S11: Weigh 500 parts of Salvia miltiorrhiza, 500 parts of Chuanxiong, and 500 parts of Pueraria root according to weight;

[0021] S12 was boiled for 1.5 hours after adding water for the first time and for 1 hour after adding water for the second time. The two decoctions were combined.

[0022] The amount of water added each time is 8-12 times the weight of the medicinal materials;

[0023] S13: filtering the decoction and concentrating it to a relative density of 1.08, and filtering it while hot;

[0024] S14: Concentrate the filtrate again to an appropriate volume, add sucrose powder to make granules,

[0025] Wherein, the amount of sucrose powder is 20%-40% of the total weight of the granules;

[0026] S15 is made into 1000g granular finished product through low temperature drying or spray drying process.

[0027] The low-temperature drying temperature is 40-60° C., the drying time is 8-12 hours, and the particle size range is 10-60 mesh.

[0028] Preferably, in the chromatographic conditions, the system suitability requirements are that the number of theoretical plates is not less than 5000 based on the puerarin peak, not less than 6000 based on the salvianolic acid B peak, the resolution is ≥2.0, and the tailing factor is 0.8-1.5.

[0029] Preferably, the quality standard in step S5 also includes: determining the content of puerarin and salvianolic acid B in Tongmai Granules, and calculating the content limit with reference to the peak area of ​​the reference solution in the fingerprint, the puerarin content is not less than 55 mg / bag, and the salvianolic acid B content is not less than 40 mg / bag.

[0030] Preferably, the quality standards also include identification and inspection items of Tongmai Granules. Identification adopts thin layer chromatography, and inspection items include particle size, moisture, filling volume difference, solubility and microbial limit.

[0031] Among them, the filling volume difference limit is ±7%, the solubility requirement is that all the substances must be dissolved within 5 minutes, and the microbial limit test is carried out in accordance with General Rules 1105 and 1106 of the 2025 edition of the "Chinese Pharmacopoeia".

[0032] Preferably, thin layer chromatography is used to identify Danshen, Chuanxiong and Pueraria root in Tongmai granules, and the specific steps include:

[0033] S61: Identification of Danshen: add ferric chloride ethanol solution to the test solution to develop color, and examine the spots under sunlight;

[0034] S62: Identification of Chuanxiong: the test solution was examined for fluorescent spots at 365 nm ultraviolet light;

[0035] S63: Identification of puerarin: examine the fluorescent spots of the test solution under ultraviolet light at 365nm and compare them with those of the reference solution.

[0036] Preferably, among the inspection items, the microbial limit inspection includes the total aerobic bacteria count, the total mold and yeast count, and the control bacteria inspection, wherein the total aerobic bacteria count is ≤600 cfu / g, the total mold and yeast count is ≤50 cfu / g, and Escherichia coli shall not be detected.

[0037] Compared with the existing technology, this medium frequency heat treatment process for supporting rollers has the following beneficial effects:

[0038] 1. The method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint provided by the present invention realizes simultaneous quantification of multiple components through high-performance liquid chromatography-fingerprint analysis, establishes an HPLC fingerprint method that simultaneously covers water-soluble components, realizes simultaneous monitoring of multiple characteristic peaks of Tongmai Granules, solves the limitations of traditional single component detection, and objectively quantifies the stability of the production process through similarity evaluation.

[0039] 2. The method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint provided by the present invention use a new detection method to analyze the proportion of each component in the drug, thereby ensuring the stability and uniformity of the drug's efficacy, further improving the Tongmai Granule product, and reducing product safety risks.

[0040] 3. The method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint provided by the present invention can quickly screen out products with process defects through the fingerprint similarity threshold, and is suitable for quality evaluation across the industry.

[0041] 4. The method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint provided by the present invention are tested by high-performance liquid chromatography, which enhances the strength of product quality description, has good batch-to-batch quality consistency, and has a more competitive advantage for the product.

[0042] In summary, the present invention provides a method for establishing the HPLC fingerprint of Tongmai Granules and its fingerprint. Through process refinement, detection technology upgrades and standard improvements, it solves the common problems of batch-to-batch quality fluctuations, insufficient efficacy of sugar-free types and safety risks of Chinese medicine granules, making the product significantly superior to competing products in terms of active ingredient content, stability and safety, providing a technical barrier for seizing the market share of cardiovascular Chinese patent medicines. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figure 1 This is the fingerprint of multiple batches of products of Sunflower Pharmaceutical Tangshan Company in the present invention;

[0044] Figure 2 for Figure 1 The similarity comparison results of the fingerprint maps;

[0045] Figure 3 This is a comparison chart of the contents of Tongmai Granules of the present invention and products from commercial manufacturers;

[0046] Figure 4 The fingerprints of Tongmai granules from different manufacturers in the present invention;

[0047] Figure 5 for Figure 4 Comparison results of similarity of fingerprint spectra of Tongmai granules from different manufacturers. DETAILED DESCRIPTION

[0048] The following are specific embodiments of the present invention and the accompanying drawings to further describe the technical solutions of the present invention, but the present invention is not limited to these embodiments.

[0049] Example 1:

[0050] The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof comprise the following steps:

[0051] S1: Take Danshen 500g, Chuanxiong 500g, Gegen 500g, decoct twice, the first time 1.5 hours, the second time 1 hour, combine the decoction, filter, concentrate the filtrate to a relative density of 1.08 (80°C), filter while hot, concentrate the filtrate to the appropriate amount, add an appropriate amount of sucrose powder, make granules, dry at low temperature, make 1000g; or spray dry, add an appropriate amount of sucrose powder, mix evenly, make granules 1000g;

[0052] S2: Gegen content determination test solution preparation: take about 0.1g of granules, accurately weigh, add 75% methanol 25ml, weigh, ultrasonic treatment (power 300W, frequency 40kHz) for 30 minutes, cool, weigh again, make up the weight loss with 75% methanol, shake well, filter, take the filtrate, and get,

[0053] Danshen content determination test solution preparation: take about 0.5g of granules, accurately weigh, add 30% ethanol 50ml, weigh, ultrasonic treatment (power 300W, frequency 40kHz) for 30min, cool, weigh again, make up the weight loss with 30% ethanol, shake well, filter, take the filtrate, and get;

[0054] S3: Gegen content determination control solution preparation: take Gegenin control substance, accurately weigh, add 30% ethanol to make a solution containing 40μg per 1ml,

[0055] Danshen content determination control solution preparation: take Danshensan B control substance, accurately weigh, add 75% methanol to make a solution containing 80μg per 1ml, and get.

[0056] S4: Chromatographic conditions:

[0057] Chromatographic column: octadecylsilane bonded silica gel chromatographic column (Agilent ZORBAX SB-C18, 4.6mm x 250mm, mobile phase A: acetonitrile-water solution (10:90) for Gegen content detection, mobile phase B: acetonitrile-0.1% phosphoric acid solution (22:78) for Danshen content detection;

[0058] Set gradient elution program;

[0059] 0-5min, mobile phase A, 90%→85%, mobile phase B, 10%→15%, flow rate: 0.8→1.0ml / min, column temperature: 30°C;

[0060] 5-15min, mobile phase A, 85%→75%, mobile phase B, 15%→25%, flow rate: 1.0→1.2ml / min, column temperature: 30°C;

[0061] 15-25 min, mobile phase A, 75% → 65%, mobile phase B, 25% → 35%, flow rate: 1.2 → 1.0 ml / min, column temperature: 30°C;

[0062] Setting parameters: detection wavelength of Pueraria lobata content: 250nm, detection wavelength of Salvia miltiorrhiza content: 286nm, column temperature: 30℃.

[0063] Accurately pipette 10 μl of each reference solution and test solution, inject into liquid chromatograph, measure, and record the chromatogram;

[0064] Determination of protocatechuic aldehyde and daidzein content: Preparation of test solution: 0.5 g of Tongmai granule powder was accurately weighed and placed in a stoppered Erlenmeyer flask; 50 ml of 70% methanol solution was accurately added and the flask was sealed; ultrasonic treatment (power 250 W, temperature 25°C) was performed for 30 minutes, and the mixture was allowed to cool and then weighed; the filtrate was filtered and the filtrate was filtered through a 0.45 μm microporous membrane to obtain the test solution;

[0065] Preparation of reference solution:

[0066] Take appropriate amount of protocatechuic aldehyde reference substance and daidzein reference substance and weigh them accurately;

[0067] Add 70% methanol to prepare a mixed reference solution containing 0.2 mg of protocatechuic aldehyde and 0.5 mg of daidzein per 1 ml;

[0068] Chromatographic conditions:

[0069] Chromatographic column: octadecylsilane bonded silica column (Agilent ZORBAX SB-C18, 4.6 mm × 250 mm, 5 μm); mobile phase: acetonitrile-0.1% phosphoric acid solution (15:85), isocratic elution; detection wavelength: protocatechuic aldehyde 280 nm, daidzein 254 nm; flow rate: 1.0 mL / min, column temperature 30°C; injection volume: 10 μL.

[0070] like Figure 1 and Figure 2 As shown, three batches of products from Sunflower Pharmaceuticals Tangshan Company were tested by HPLC to generate fingerprints. The "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" was used for analysis, and the similarities were all ≥0.95. The fingerprints of multiple batches of products from Sunflower Pharmaceuticals Tangshan Company show that they begin with a high-intensity dead time peak, then go through a characteristic region containing multiple compound peaks (manifested as response value fluctuations), and finally return to zero response value. The fingerprints of 10 batches of products show a high degree of consistency in the characteristic peak patterns of these 10 batches on the fingerprints. Good consistency indicates that the chemical composition and relative content of these 10 batches of products are stable, and the production process is well controlled.

[0071] like Figures 1-5As shown in the figure, 37 batches of samples from 9 companies (including 10 batches from Sunflower Pharmaceutical Tangshan Company and 27 batches from competitors) were selected. Puerarin, salvianolic acid B, daidzein, and protocatechuic aldehyde were detected by HPLC. The similarity between batches was calculated based on 12 characteristic peaks. The results are as follows:

[0072] Table 1 Comparative statistics of the main components of Tongmai granules from multiple manufacturers

[0073]

[0074] The results in Table 1 show that the content of main ingredients of Kuihua Pharmaceutical Tangshan Company is significantly higher than the industry average, and the effective ingredient retention rate is 2.1-3.5 times higher than that of sugar-free granules, confirming that differences in process control directly affect the ingredient transfer rate.

[0075] The results in Table 2 show that in the fingerprint study, it can be judged that all manufacturers use water extraction technology for preparation. Due to different process controls, the extraction transfer rates between components are not the same. In general, Taiji Group, Sunflower Pharmaceutical Tangshan Company, and Shenwei Pharmaceutical are better than other companies in various indicators. From the perspective of uniformity control of multiple batches of products, these three companies are also significantly better than other companies' products, especially the main ingredients such as puerarin, salvianolic acid B, daidzein, protocatechuic aldehyde and other active ingredients. The products of these three companies are also ahead of other companies, and will have better effects in pharmacology and efficacy.

[0076] Table 2 Comparison of the main components of Tongmai granules from different manufacturers

[0077]

[0078]

[0079] Conclusion: Sunflower Pharmaceutical Tangshan Company surpasses competitors by more than 40% in core ingredients such as puerarin (98.2 mg / bag) and salvianolic acid B (58.7 mg / bag), with fingerprint similarity ≥0.96 (SD ≤ 2.1%), ensuring consistency in clinical efficacy. The HPLC fingerprint method can effectively distinguish high-quality products (similarity >0.90) from products with process defects. By establishing an HPLC fingerprint method that simultaneously covers water-soluble components, simultaneous monitoring of multiple characteristic peaks of Tongmai Granules is achieved, overcoming the limitations of traditional single ingredient detection.

[0080] Example 2: Based on Example 1,

[0081] The quality standards also include identification and inspection items of Tongmai Granules. Identification adopts thin layer chromatography, and inspection items include particle size, moisture, filling volume difference, solubility and microbial limit.

[0082] Among them, the filling volume difference limit is ±7%, the solubility requirement is that all the substances must be dissolved within 5 minutes, and the microbial limit test is carried out in accordance with General Rules 1105 and 1106 of the 2025 edition of the "Chinese Pharmacopoeia".

[0083] Thin layer chromatography was used to identify Danshen, Chuanxiong and Pueraria root in Tongmai granules. The specific steps include:

[0084] To identify Danshen, take 5g of this product, grind it into powder, add 50ml of ethanol, sonicate for 25 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 20ml of water, add 2 drops of hydrochloric acid, and extract twice with ethyl acetate by shaking, 30ml each time. Combine the ethyl acetate solutions, evaporate to dryness at low temperature, and dissolve the residue in 1ml of methanol to prepare the test solution. Take another 2g of Danshen control medicinal material, add 30ml of water, decoct for 30 minutes, filter, and add 2 drops of hydrochloric acid to the filtrate. Prepare the control medicinal material solution in the same way. According to the thin layer chromatography method (General Rules 0502 of the 2020 edition of the Chinese Pharmacopoeia), take 3μl of each of the above two solutions and spot them on the same silica gel G thin layer plate. Use chloroform-ethyl acetate-toluene-formic acid (24:20:10:6) as the developing solvent, develop, remove, dry, spray with 1% ferric chloride ethanol solution, and inspect under sunlight. In the chromatogram of the test sample, spots of the same color appear at the corresponding positions in the chromatogram of the control medicinal material;

[0085] To identify Chuanxiong, take 3g of this product, add 30ml of water, dissolve it with slight heat, filter, transfer the filtrate to a separatory funnel, add 60ml of ether and 2ml of saturated sodium chloride solution, shake gently, let it stand, and separate the ether solution (if emulsified, separate the emulsion layer, add 1ml of anhydrous ethanol, shake well, and separate the ether solution). Continue to extract the aqueous layer with 60ml of ether, combine the ether solutions, evaporate to dryness at low temperature, and dissolve the residue in 0.5ml of methanol to prepare the test solution. Take another 2g of Chuanxiong control medicinal material, add 40ml of water, decoct for 15 minutes, filter, cool, transfer to a separatory funnel, add 60ml of ether, and prepare the control medicinal material solution in the same way. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 General Chapter 0502), 5 μl of the test solution and 8 μl of the control herbal solution were respectively spotted as strips on the same silica gel G thin layer plate. Ether-n-hexane (9:1) was used as the developing solvent. The plate was developed, removed, dried, and examined under ultraviolet light (365 nm). In the chromatogram of the test sample, a fluorescent spot of the same color appeared at the corresponding position in the chromatogram of the control herbal drug;

[0086] For puerarin identification, take the test solution under the Danshen identification item as the test solution. Take another puerarin reference substance, add methanol to make a solution containing 0.5 mg per 1 ml, and use it as the reference solution. According to the thin layer chromatography method (Chinese Pharmacopoeia 2020 Edition General Rules 0502), take 2 μl of each of the above two solutions and spot them in strips on the same silica gel G thin layer plate, use chloroform-methanol-water (7:2.5:0.25) as the developing agent, pre-saturate for 15 minutes, develop, take out, dry, and inspect under ultraviolet light (365 nm). In the test sample chromatogram, fluorescent spots of the same color appear at the corresponding position of the reference chromatogram.

[0087] Among the inspection items, microbial limit inspection includes total aerobic bacteria, total mold and yeast count, and control bacteria inspection, among which the total aerobic bacteria count is ≤600cfu / g, the total mold and yeast count is ≤50cfu / g, and Escherichia coli must not be detected.

[0088] Table 3 Comparison results between this product and the legal standard of Tongmai Granules

[0089]

[0090] Conclusion: The inspection items of this product are the same as the legal standards, and the control limits are stricter than the legal standards; the quality standards of Tongmai Granules containing sucrose on the market are all consistent.

[0091] Table 4 Comparative results of properties analysis of Tongmai granules from multiple manufacturers

[0092]

[0093]

[0094] The results in Table 4 show that, through a comparison of Tongmai Granules from several manufacturers, among the samples collected, with the exception of Hebei Wansui Pharmaceutical, which uses a wet granulation process, all other manufacturers use a one-step granulation method. Taiji is more hygroscopic, while the hygroscopicity of other manufacturers varies slightly, but overall, is similar. Most manufacturers of sugar-free Tongmai Granules use a wet granulation process, with significant variations in process parameters and distinct granule color. This difference was also evident in subsequent testing, demonstrating the quality advantage of this product's sugar-containing Tongmai Granules over its sugar-free counterpart.

[0095] Conclusion: According to the above method, the fingerprints of puerarin, tanshinone B, protocatechuic aldehyde and daidzein of this product, Jilin Ruilong Pharmaceutical, Henan Xingyuan Pharmaceutical, Jilin Jierji Pharmaceutical, Xiuzhen Pharmaceutical, Harbin Dayang Pharmaceutical, Hebei Wansui Pharmaceutical, Taiji Group Chongqing Pharmaceutical Factory No. 2 and Shenwei Pharmaceutical were tested. The results showed that the quality of Tongmai Granules of this product was good, the batch-to-batch quality consistency of daidzein and protocatechuic aldehyde in the product was good, and the content of daidzein and protocatechuic aldehyde was also high.

[0096] The specific embodiments described herein are merely illustrative of the spirit of the present invention. Persons skilled in the art may make various modifications, additions, or substitutions to the described specific embodiments without departing from the spirit of the present invention or exceeding the scope of the appended claims.

Claims

1. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof are characterized in that: The following steps are involved: S1: Prepare the test solution. Take 0.1-0.5g of Tongmai granule powder, accurately weigh it, add 25ml of 30% ethanol or 70% methanol solution, and ultrasonically treat it for 30min at a power of 300W and a frequency of 40kHz. After cooling, make up the weight loss, and filter it through a 0.45μm filter membrane to obtain the test solution. Among them, Tongmai Granules include the following ingredients in terms of parts: 500 parts of Danshen, 500 parts of Chuanxiong, and 500 parts of Pueraria; S2: Take puerarin and salvianolic acid B reference substances and add solvent to make a solution containing 0.5 mg per 1 ml; S3: Select chromatographic conditions: S31: Chromatographic column: octadecylsilane bonded silica gel column (Agilent ZORBAX SB-C18, 4.6 mm × 250 mm, 5 μm); S32: mobile phase A: acetonitrile-water solution (10:90), mobile phase B: acetonitrile-0.1% phosphoric acid solution (22:78); S33: set gradient elution program; S34: Set parameters: detection wavelength: 250 nm or 286 nm, flow rate: 1.0 mL / min, column temperature: 30°C, injection volume: 10 μL; S3: Inject the test solution and reference solution separately and record the chromatogram; S4: Similarity evaluation is performed using the "Chinese Herbal Medicine Chromatographic Fingerprint Similarity Evaluation System", and the similarity must not be less than 0.90; S5: Test the Tongmai Granules to be tested, evaluate the results by the similarity of the fingerprint patterns, and determine whether the quality meets the requirements according to the quality standards.

2. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof as claimed in claim 1, characterized in that: The preparation of the Tongmai granules comprises the following steps: S11: Weigh 500 parts of Salvia miltiorrhiza, 500 parts of Chuanxiong, and 500 parts of Pueraria root according to weight; S12 was boiled for 1.5 hours after adding water for the first time and for 1 hour after adding water for the second time. The two decoctions were combined. The amount of water added each time is 8-12 times the weight of the medicinal materials; S13: filtering the decoction and concentrating it to a relative density of 1.08, and filtering it while hot; S14: Concentrate the filtrate again to an appropriate volume, add sucrose powder to make granules, Wherein, the amount of sucrose powder is 20%-40% of the total weight of the granules; S15 is made into 1000g granular finished product through low temperature drying or spray drying process. The low-temperature drying temperature is 40-60° C., the drying time is 8-12 hours, and the particle size range is 10-60 mesh.

3. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof as claimed in claim 1 or 2, characterized in that: Among the chromatographic conditions, the system suitability requirements are that the number of theoretical plates is not less than 5000 based on the puerarin peak, not less than 6000 based on the salvianolic acid B peak, the resolution is ≥2.0, and the tailing factor is 0.8-1.

5.

4. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof as claimed in claim 1, characterized in that: The quality standard in step S5 also includes: determining the content of puerarin and salvianolic acid B in Tongmai Granules, and calculating the content limit with reference to the peak area of ​​the reference solution in the fingerprint, and the puerarin content is not less than 55 mg / bag, and the salvianolic acid B content is not less than 40 mg / bag.

5. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof as claimed in claim 4, characterized in that: The quality standards also include identification and inspection items of Tongmai Granules. Identification adopts thin layer chromatography, and inspection items include particle size, moisture, filling volume difference, solubility and microbial limit. Among them, the filling volume difference limit is ±7%, the solubility requirement is that all the substances must be dissolved within 5 minutes, and the microbial limit test is carried out in accordance with General Rules 1105 and 1106 of the 2025 edition of the "Chinese Pharmacopoeia".

6. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof as claimed in claim 5, characterized in that: The thin layer chromatography method for identifying Danshen, Chuanxiong and Puerariae in Tongmai granules specifically comprises the following steps: S61: Identification of Danshen: add ferric chloride ethanol solution to the test solution to develop color, and examine the spots under sunlight; S62: Identification of Chuanxiong: the test solution was examined for fluorescent spots at 365 nm ultraviolet light; S63: Identification of puerarin: examine the fluorescent spots of the test solution under ultraviolet light at 365nm and compare them with those of the reference solution.

7. The method for establishing the HPLC fingerprint of Tongmai Granule and the fingerprint thereof according to any one of claims 5 or 6, characterized in that: Among the inspection items mentioned above, the microbial limit inspection includes the total aerobic bacteria count, the total mold and yeast count, and the control bacteria inspection, among which the total aerobic bacteria count is ≤600 cfu / g, the total mold and yeast count is ≤50 cfu / g, and Escherichia coli shall not be detected.

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