A method for establishing UPLC fingerprint of Weimaining capsules

Abstract

The present invention discloses a method for establishing a UPLC fingerprint of Weimaining capsules, comprising the following steps: (1) preparing a test solution; (2) respectively examining the precision of an ultra-high performance liquid chromatograph, the repeatability of a test solution preparation method, and the stability of the test solution; (3) subjecting the test solution to gradient elution in a chromatographic column, and scanning the test solution using an ultra-high performance liquid chromatograph to obtain a chromatogram of the test solution; (4) identifying the common peak components of the chromatogram using reference substances such as gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin. (5) establishing a fingerprint. The ultra-high performance liquid chromatograph used in the present invention has high precision, and the test solution preparation method provided has good repeatability. The fingerprint established based on the above two points has strong stability, and the speed of establishing the fingerprint is fast, the sensitivity is high, and the separation of the chromatographic peaks is good.

Description

Technical Field

[0001] The present invention relates to the technical field of drug detection, and in particular to a method for establishing a UPLC fingerprint of Weimaining capsules. Background Art

[0002] Existing quality standards for Chinese herbal medicine slices, compound decoctions, and granules generally include the following aspects: morphological identification, physical and chemical identification, and spectral chromatography analysis, among which spectral chromatography analysis generally includes thin-layer chromatography, high-performance liquid chromatography content determination, fingerprint analysis, etc. As a single-ingredient Chinese herbal preparation, Weimaining Capsules have relatively prescribed processing and usage methods. However, due to their relatively complex chemical composition, multi-target and multi-level pharmacological effects, and numerous interfering factors, the quality and process of Weimaining Capsules, from raw materials to finished products, are difficult to accurately control, making it difficult to fully reflect the quality status of Weimaining Capsules.

[0003] Fingerprinting is an effective comprehensive analytical method that uses modern analytical techniques to identify the authenticity of traditional Chinese medicines (TCMs) or Chinese patent medicines (TCMs), and to evaluate the consistency and stability of the quality of raw materials, semi-finished products, and finished products. Compared with existing identification methods based on single active ingredients or indicator components, TCM fingerprinting technology integrates a variety of effective chemical information, resulting in more scientific and reasonable results. TCM fingerprinting has become a recognized and important method for TCM quality control both domestically and internationally. The research and establishment of fingerprinting is of great significance for improving the quality of TCM and promoting its modernization. Ultra-high performance liquid chromatography (UPLC), with its advantages of high sensitivity, high efficiency, and good stability, is a widely used method for detecting TCM fingerprints. It can separate multiple components in a short time with good resolution. Currently, no studies have used UPLC to establish a fingerprinting method for Weimaining and identify its components. Therefore, the establishment of a fingerprint for Weimaining capsules using UPLC is of great significance for quality control analysis of Weimaining capsules. Summary of the Invention

[0004] The present invention provides a method for establishing a UPLC fingerprint of Weimaining capsules, comprising:

[0005] (1) Preparation of test solution:

[0006] Take 2g of the contents of Weimaining Capsules, place it in a stoppered conical flask, add 50ml of water, heat under reflux for 20-40 minutes, cool, filter, concentrate the filtrate under reduced pressure, add 50ml of dilute ethanol, heat under reflux for 20-40 minutes, take it out, cool, filter, take 25ml of the filtrate, concentrate under reduced pressure, wash the residue with acetonitrile-water mixed solution several times, transfer the washing solution to a 10ml measuring flask, dilute to the mark with acetonitrile-water mixed solution, shake well, centrifuge, take 5ml of the supernatant, add it to a polyamide column, elute with 50ml of water, discard the water, and then elute with 200ml of ethanol, collect the eluate, concentrate under reduced pressure, dissolve the residue with acetonitrile-water mixed solution, transfer it to a 10ml measuring flask, dilute to 10ml with acetonitrile-water mixed solution, shake well, filter, and take the filtrate as the test solution;

[0007] (2) respectively examine the precision of the ultra-high performance liquid chromatography instrument, the repeatability of the test solution preparation method, and the stability of the test solution;

[0008] (3) gradient eluting the test solution in a chromatographic column and scanning the test solution using an ultra-high performance liquid chromatograph to obtain a chromatogram of the test solution;

[0009] (4) Use the reference substances gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin to identify the common peak components in the chromatogram;

[0010] (5) Establishing fingerprint map.

[0011] Furthermore, the fingerprint is established by the following steps:

[0012] Take 15 batches of Weimaining capsules to prepare test solutions according to step (1), perform gradient elution in a chromatographic column, and scan the test solutions using an ultra-high performance liquid chromatograph to obtain chromatograms of the 15 batches of test solutions;

[0013] Based on the chromatograms of 15 batches of test sample solutions, the median method was used with a time window width of 0.1, and the control fingerprints of the 15 batches of Weimaining Capsules were automatically matched according to the pre-set sample reference maps.

[0014] Furthermore, the polyamide column has a mesh size of 30-60, an inner diameter of 1.0 cm, and a column length of 15 cm.

[0015] Furthermore, the centrifugal speed is 3000 revolutions per minute, and the centrifugal time is 5 minutes.

[0016] Furthermore, the heating reflux time is 30 minutes.

[0017] Furthermore, the ratio of acetonitrile to water is 10:90.

[0018] Furthermore, the chromatographic column is: 1.6 μm, 2.1*150 mm; the column temperature is: 30° C.; the mobile phase is: acetonitrile-0.1% phosphoric acid solution; the flow rate is: 0.3 mL / min; the detection wavelength is: 278 nm; and the injection volume is: 3 μL.

[0019] Furthermore, the gradient elution value is:

[0020] timemin(0, 10, 17, 25, 30, 35, 40);

[0021] Acetonitrile (7%, 8%, 10%, 12%, 12%, 14%, 14%);

[0022] 0.1% phosphoric acid (93%, 92%, 90%, 88%, 88%, 86%, 86%).

[0023] The present invention also provides a UPLC fingerprint of Weimaining Capsules, which is obtained according to the above-mentioned method for establishing the UPLC fingerprint of Weimaining Capsules.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] The ultra-high-performance liquid chromatograph used in the present invention has high precision, and the provided method for preparing the test solution has good reproducibility. Based on these two points, the fingerprint spectrum established is more stable. At the same time, compared with the HPLC fingerprint spectrum establishment method, the present invention establishes the fingerprint spectrum faster, has higher sensitivity, and has better separation of chromatographic peaks. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 The chromatogram of the test sample obtained by heating and refluxing through a polyamide column in the present invention;

[0027] Figure 2 This is a chromatogram of the test sample obtained by ultrasound using 10% acetonitrile in the present invention;

[0028] Figure 3 This is a chromatogram of the test sample obtained by ultrasonication with 50% methanol in the present invention;

[0029] Figure 4 This is a schematic diagram of the chromatographic peaks of the test sample and the reference sample when the extraction time is 20 minutes;

[0030] Figure 5 This is a schematic diagram of the chromatographic peaks of the test sample and the reference sample when the extraction time is 30 minutes;

[0031] Figure 6 Schematic diagram of the chromatographic peaks of the test sample and the reference sample when the extraction time is 40 min;

[0032] Figure 7Schematic diagram of the chromatographic peaks of the test sample and reference sample prepared by the heating reflux method of the present invention;

[0033] Figure 8 Schematic diagram of the chromatographic peaks of the test sample and reference sample prepared by ultrasonic dissolution method of the present invention;

[0034] Figure 9 This is the control fingerprint of 15 batches of test products of the present invention;

[0035] Figure 10 The chromatograms of 15 batches of test products of the present invention are as follows;

[0036] Figure 11 is a chromatogram of the mixed reference solution of the present invention;

[0037] Figure 12 It is a chromatogram of the test solution of the present invention;

[0038] Figure 13 The chromatogram of the test solution obtained by eluting with gradient A of the present invention;

[0039] Figure 14 The chromatogram of the test solution obtained by eluting with gradient B of the present invention;

[0040] Figure 15 The chromatogram of the test solution obtained by elution with a C gradient of the present invention;

[0041] Figure 16 The chromatographic column of the present invention is Chromatogram of the test solution obtained at T3 (1.6 μm, 2.1*150 mm);

[0042] Figure 17 The chromatographic column of the present invention is ACQUITY Chromatogram of the test solution obtained at HSS T3 (1.8 μm, 2.1*150 mm);

[0043] Figure 18 The chromatographic column of the present invention is Chromatogram of the test solution obtained at (1.7 μm, 150*2.1 mm);

[0044] Figure 19 This is a chromatogram of the test solution obtained when the flow rate is 0.2 mL / min;

[0045] Figure 20 This is a chromatogram of the test solution obtained when the flow rate is 0.3 mL / min;

[0046] Figure 21 This is a chromatogram of the test solution obtained when the flow rate is 0.4 mL / min;

[0047] Figure 22 The chromatogram of the test solution obtained when the column temperature of the present invention is 25°C;

[0048] Figure 23 The chromatogram of the test solution obtained when the column temperature of the present invention is 30°C;

[0049] Figure 24 The chromatogram of the test solution obtained when the column temperature of the present invention is 35°C. DETAILED DESCRIPTION

[0050] In order to make the purpose, technical solution and advantages of this technical solution more clear, the following technical solution is further described in detail in conjunction with specific implementation methods. It should be understood that these descriptions are only exemplary and are not intended to limit the scope of this technical solution.

[0051] First, the instruments and reagents used in the present invention are introduced:

[0052] Waters ultra-high performance liquid chromatograph, KQ-250DE CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.), rotary evaporator RV3E0 (IKA), acetonitrile (F240907M201, chromatographic grade, Fisher, USA), phosphoric acid (analytical grade, Sinopharm Chemical Reagent Co., Ltd.), pure water (Wahaha), anhydrous ethanol (analytical grade, Sinopharm Chemical Reagent Co., Ltd.), methanol (chromatographic grade, Fisher, USA), polyamide (30-60 mesh, Changfeng Chemical Co., Ltd.).

[0053] Next, the samples used in the present invention are introduced:

[0054] Weimaining Capsules (batch number 230802, Huayi Pharmaceutical Co., Ltd.), epicatechin (110878-201703, China Food and Drug Inspection Institutes), proanthocyanidin B2 (112089-202201, China Food and Drug Inspection Institutes), gallic acid (110831-202407, China Food and Drug Inspection Institutes), protocatechuic acid (15244, Shanghai Standard Standards Technical Services Co., Ltd.), protocatechuic aldehyde (102752920, Shanghai MacLean Biochemical Technology Co., Ltd.), rutin (100080-202012, China Food and Drug Inspection Institutes).

[0055] Next, the method for establishing the UPLC fingerprint of Weimaining Capsules provided by the present invention is introduced from the following aspects:

[0056] (1) Preparation of test solution:

[0057] Take 2g of the contents of Weimaining capsules, place them in a stoppered conical flask, add 50ml of water, heat and reflux for 30 minutes, cool, filter, and concentrate the filtrate under reduced pressure to near dryness. Add 50ml of dilute ethanol, heat and reflux for 30 minutes, take out, cool, filter, take 25ml of the filtrate, and concentrate under reduced pressure to near dryness. Wash the residue with a mixed solution of acetonitrile and water (10:90) in several times. Transfer the washing solution to a 10ml volumetric flask, add a mixed solution of acetonitrile and water (10:90) to dilute to the scale, shake well, and centrifuge (speed of 300 per minute). 00 rpm) for 5 minutes, take 5 ml of the supernatant, add it to a polyamide column (30-60 mesh, inner diameter 1.0 cm, column length 15 cm, wet packing), elute with 50 ml of water, discard the water, and then elute with 200 ml of ethanol, collect the eluate, and concentrate under reduced pressure to near dryness, dissolve the residue in acetonitrile-water (10:90) mixed solution, transfer it to a 10 ml volumetric flask, add acetonitrile-water (10:90) mixed solution to dilute to 10 ml, shake well, filter, and take the filtrate as the test solution.

[0058] The above-mentioned method for preparing the test solution was obtained through multiple comparative experiments. The specific experimental process is as follows:

[0059] First, the present invention conducted comparative experiments on the heating reflux method and the ultrasonic dissolution method, and obtained the following results: Figure 7-8 The schematic diagram of the chromatographic peaks of the test sample and reference sample prepared by the heating reflux method and the ultrasonic dissolution method is shown. Among them, the chromatographic peak area of ​​the test sample solution prepared by the heating reflux method is 1035856, and the chromatographic peak area of ​​the test sample solution prepared by the ultrasonic dissolution method is 719997. The results show that the chromatographic peak area of ​​the test sample solution extracted by the heating reflux method is larger and the extraction rate is high, so the heating reflux method is used for extraction.

[0060] Secondly, the present invention conducted comparative experiments on different extraction times of the heating reflux method, and obtained the following results: Figure 4-6 The following is a schematic diagram of the chromatographic peaks of the test sample and the reference sample at extraction times of 20 min, 30 min, and 40 min. Figure 4 As shown in FIG, when heated under reflux for 20 minutes, the chromatographic peak area is 376297. Figure 5 As shown in FIG, when heated under reflux for 30 minutes, the chromatographic peak area is 1035856. Figure 6 As shown in the figure, when heated under reflux for 40 minutes, the chromatographic peak area is 492753. The results show that the peak area obtained by extraction for 30 minutes is larger, so the extraction time is set to 30 minutes.

[0061] Again, the present invention provides two other comparative examples to verify the effectiveness of the test solution preparation method of the present invention, specifically:

[0062] Comparative Example 1: Take about 0.4g of the capsule contents, accurately weigh it, place it in a conical flask, accurately add 50mL of 10% acetonitrile, dissolve it by ultrasonication, let it stand for 1h, then heat it under reflux for 1h, let it cool, weigh it again, make up the loss with 10% acetonitrile, shake it well, and take the filtrate as the test solution. And obtain the following Figure 2 Shown is the chromatogram of the test product obtained by sonication with 10% acetonitrile.

[0063] Comparative Example 2: Take about 0.4g of the capsule contents, weigh accurately, place in a conical flask, add 50mL of 50% methanol, dissolve by ultrasonication, cool, shake well, and take the filtrate as the test solution. Figure 3 The chromatogram shown is of the test product obtained by sonication in 50% methanol.

[0064] Also, see Figure 1 The chromatogram of the test sample obtained by heating and reflux through the polyamide column is compared with Figure 1-3 It can be seen that the chromatographic peaks obtained by the polyamide column separation method have a stable peak baseline, more detected peaks, better peak shape, and better resolution of most peaks. This confirms the effectiveness of the test solution preparation method disclosed in the present invention.

[0065] (2) The precision of the ultra-high performance liquid chromatography instrument, the repeatability of the test solution preparation method and the stability of the test solution were examined respectively.

[0066] Preparation of reference solution:

[0067] Take an appropriate amount of epicatechin reference substance, accurately weigh it, place it in a 10 mL volumetric flask, add 50% methanol and ultrasonically dissolve it to the scale to prepare a reference substance solution containing 1 mg of epicatechin per 1 mL.

[0068] Investigating the precision of ultra-high performance liquid chromatography:

[0069] The same test solution was injected six times. Using epicatechin as the reference peak (S), the relative standard deviations (RSDs) of the relative retention times and relative peak areas of the remaining common peaks were calculated. The results showed that the RSDs for the relative retention times of the common peaks were <1% (Table 1) and <2.5% (Table 2), indicating good instrument precision.

[0070] Table 1. Relative retention time of some chromatographic peaks investigated for precision

[0071]

[0072] Table 2. Relative peak areas of some chromatographic peaks investigated for precision

[0073]

[0074] Investigate the repeatability of the test solution preparation method:

[0075] Six portions of Weimaining capsules from the same batch were accurately weighed and test solutions were prepared according to the test solution preparation method disclosed in (1) above. Samples were injected separately, and epicatechin was used as the reference peak (S). The RSDs of the relative retention times and relative peak areas of the remaining common peaks were calculated. The results showed that the RSDs of the relative retention times of the common peaks were <2% (Table 3) and the RSDs of the relative peak areas were <5% (Table 4), indicating that the test solution preparation method had good reproducibility.

[0076] Table 3 Relative retention time of some chromatographic peaks in repeatability study

[0077]

[0078] Table 4 Relative peak areas of some chromatographic peaks examined for repeatability

[0079]

[0080] Investigate the stability of the test solution:

[0081] The same test solution was injected at 0, 2, 4, 6, 8, 12, 24, and 48 h, respectively. Taking epicatechin as the reference peak (S), the RSDs of the relative retention times and relative peak areas of the remaining common peaks were calculated. The results showed that the RSDs of the relative retention times of the common peaks were between 0.32% and 0.66%, and the RSDs of the relative peak areas were between 0.73% and 3.21%, indicating that the test solution was stable within 48 h.

[0082] Table 5 Relative retention time of some chromatographic peaks investigated for stability

[0083]

[0084]

[0085] Table 6 Relative peak areas of some chromatographic peaks investigated for stability

[0086]

[0087] (3) Gradient elution of the test solution in a chromatographic column is performed, and the test solution is scanned using an ultra-high performance liquid chromatograph to obtain a chromatogram of the test solution.

[0088] Here, the embodiment of the present invention determines the parameters of the chromatographic column by a controlled variable method.

[0089] First, the present invention conducted comparative experiments on different types of chromatographic columns, and obtained the following results: Figure 16-18 The chromatographic columns shown are T3 (1.6μm, 2.1*150mm), HSST3 (1.8μm, 2.1*150mm) and The chromatogram of the test solution obtained by (1.7μm, 150*2.1mm). By comparing the baseline stability of the chromatographic peak and the separation effect, the chromatographic column model is preferably T3 (1.6 μm, 2.1*150 mm).

[0090] Secondly, the embodiment of the present invention is directed to The T3 (1.6 μm, 2.1*150 mm) column was used for a comparative experiment at different column temperatures. Figure 22-24 The chromatograms of the test solution obtained when the column temperature is 25℃, 30℃ and 35℃ respectively. Figure 22-24 As shown, the column temperature is preferably 30°C.

[0091] Again, the present invention uses acetonitrile-0.1% phosphoric acid solution as the mobile phase, and conducts comparative experiments on different flow rates of the mobile phase, and obtains the following results: Figure 19-21 The chromatograms of the test solution obtained at flow rates of 0.2 mL / min, 0.3 mL / min, and 0.4 mL / min are shown. Figure 19-21 As shown, the flow rate is preferably 0.3 mL / min.

[0092] It should be noted that the detection wavelength of the above experiments was set to 278 nm and the injection volume was set to 3 μL.

[0093] Finally, the present invention conducted comparative experiments on different gradient elution values, and obtained the following results: Figure 13-15 The chromatogram of the test solution obtained by elution with gradients A, B, and C is shown. The specific gradient elution values ​​are set as follows:

[0094] A gradient: time min (0, 10, 17, 25, 30, 35, 40);

[0095] Acetonitrile (7%, 8%, 10%, 12%, 12%, 14%, 14%);

[0096] 0.1% phosphoric acid (93%, 92%, 90%, 88%, 88%, 86%, 86%).

[0097] B gradient: time min (0, 10, 17, 25, 30, 35, 40);

[0098] Acetonitrile (5%, 6%, 8%, 10%, 10%, 12%, 12%);

[0099] 0.1% phosphoric acid (95%, 94%, 92%, 90%, 90%, 88%, 88%).

[0100] C gradient: time min (0, 10, 17, 25, 30, 35, 40);

[0101] Acetonitrile (9%, 10%, 12%, 14%, 14%, 16%, 16%);

[0102] 0.1% phosphoric acid (91%, 90%, 88%, 86%, 86%, 84%, 84%).

[0103] pass Figure 13-15 It can be seen from the peak time and separation effect in the chromatogram that the gradient elution value is preferably A gradient.

[0104] (IV) Use the reference substances gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin to identify the common peak components in the chromatogram.

[0105] Preparation of reference solution:

[0106] Accurately weigh appropriate amounts of gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin into a 10 mL brown volumetric flask. Dissolve rutin in methanol to the same volume. Dissolve the remaining reference substances in 50% methanol, adjust the volume, and shake well. Take 200 μL of each of the above reference substance solutions and mix them evenly to prepare the mixed reference substance solution.

[0107] Identification of common peak components in chromatograms:

[0108] The single reference solution, mixed reference solution and test solution were scanned in the range of 210-400 nm using a PDA detector to identify the components. Figure 11-12 By comparing the chromatograms of the mixed reference solution and the test solution shown, it can be seen that the relative retention times of peaks 1, 2, 3, 4, 5, and 6 in Weimaining Capsules are consistent with the retention times of gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, and rutin reference substances, respectively.

[0109] (5) Establishing fingerprint map.

[0110] Take 15 batches of Weimaining capsules to prepare test solutions according to step (1), perform gradient elution in a chromatographic column, and scan the test solutions using an ultra-high performance liquid chromatograph to obtain chromatograms of the 15 batches of test solutions;

[0111] Based on the chromatograms of 15 batches of test sample solutions, the median method was used with a time window width of 0.1, and the control fingerprints of the 15 batches of Weimaining Capsules were automatically matched according to the pre-set sample reference maps.

[0112] Here, the Chinese medicine chromatographic fingerprint similarity evaluation system can be used to generate a control fingerprint.

[0113] As an example, 15 batches of Weimaining capsules were used to prepare test solutions, which were measured and analyzed under chromatographic conditions. The 2012 edition of the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" software was used to match the chromatograms. The sample S1 spectrum was set as the reference spectrum, and the control spectrum was generated using the median method with a time window width of 0.1. Automatic matching was performed to generate a control fingerprint spectrum.

[0114] Furthermore, the generated control fingerprint is used to evaluate the stability of the quality of the Weimaining capsule sample. Specifically, the similarity between the UPLC spectrum of each batch and its control fingerprint is calculated. Figure 9-10 The control fingerprints of 15 batches of test samples and the chromatograms of 15 batches of test samples are shown in Table 7. Further, the similarity between the chromatograms of each batch of test samples and their control fingerprints is calculated, and the similarity experimental results of 15 batches of test samples are obtained as shown in Table 7. As shown in Table 7, the similarities are all greater than 0.9, which shows that the sample quality is relatively stable as a whole.

[0115] Table 7 Similarity test results of 15 batches of test products

[0116]

[0117]

[0118] The above content is only a preferred embodiment of the present invention. For ordinary technicians in this field, many changes can be made in the specific implementation methods and application scope based on the ideas of the present technical content. As long as these changes do not deviate from the concept of the present invention, they all fall within the scope of protection of this patent.

Claims

1. A method for establishing a UPLC fingerprint of Weimaining Capsules, characterized in that: The following steps are involved: (I) Preparation of test solution: Take 2g of the contents of Weimaining Capsules, place it in a stoppered conical flask, add 50ml of water, heat under reflux for 20-40 minutes, cool, filter, concentrate the filtrate under reduced pressure, add 50ml of dilute ethanol, heat under reflux for 20-40 minutes, take it out, cool, filter, take 25ml of the filtrate, concentrate under reduced pressure, wash the residue with acetonitrile-water mixed solution several times, transfer the washing solution to a 10ml measuring flask, dilute to the mark with acetonitrile-water mixed solution, shake well, centrifuge, take 5ml of the supernatant, add it to a polyamide column, elute with 50ml of water, discard the water, and then elute with 200ml of ethanol, collect the eluate, concentrate under reduced pressure, dissolve the residue with acetonitrile-water mixed solution, transfer it to a 10ml measuring flask, dilute to 10ml with acetonitrile-water mixed solution, shake well, filter, and take the filtrate as the test solution; (2) Investigate the precision of the ultra-high performance liquid chromatography instrument, the repeatability of the test solution preparation method, and the stability of the test solution respectively; (3) Gradient elution of the test solution in a chromatographic column and scanning the test solution using an ultra-high performance liquid chromatograph to obtain a chromatogram of the test solution; (IV) Use reference substances such as gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin to identify the common peak components in the chromatogram; (5) Establishing fingerprint map; The chromatographic column used was a CORTECS® UPLC® T3 column (1.6 μm, 2.1 x 150 mm); the column temperature was 30°C; the mobile phase was acetonitrile-0.1% phosphoric acid solution; the flow rate was 0.3 mL / min; the detection wavelength was 278 nm; and the injection volume was 3 μL. The gradient elution values ​​are: The times are 0 min, 10 min, 17 min, 25 min, 30 min, 35 min, and 40 min respectively; The proportions of acetonitrile were 7%, 8%, 10%, 12%, 12%, 14%, and 14%; The proportions of 0.1% phosphoric acid are 93%, 92%, 90%, 88%, 88%, 86% and 86% respectively.

2. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: Create a fingerprint by following these steps: Prepare test solutions of 15 batches of Weimaining capsules according to step (1), perform gradient elution on a chromatographic column, and scan the test solutions using an ultra-high performance liquid chromatograph to obtain chromatograms of the 15 batches of test solutions; Based on the chromatograms of 15 batches of test sample solutions, the median method was used with a time window width of 0.1, and the control fingerprints of the 15 batches of Weimaining Capsules were automatically matched according to the pre-set sample reference maps.

3. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: The polyamide column has a mesh size of 30-60, an inner diameter of 1.0 cm, and a length of 15 cm.

4. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: The centrifugal speed was 3000 revolutions per minute and the centrifugation time was 5 minutes.

5. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: The heating reflux time was 30 minutes.

6. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: The ratio of acetonitrile to water was 10:

90.

7. A UPLC fingerprint of Weimaining Capsules, characterized by: Derived from the establishment method according to any one of claims 1 to 6.

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