Method for establishing Weimigning capsule UPLC (Ultra Performance Liquid Chromatography) fingerprint spectrum

Abstract

The invention discloses a method for establishing a Weimigning capsule UPLC (Ultra Performance Liquid Chromatography) fingerprint spectrum. The method comprises the following steps: (1) preparing a test solution; (2) respectively investigating the precision of an ultra performance liquid chromatograph, the repeatability of the preparation method of the test solution and the stability of the test solution; (3) carrying out gradient elution on the test sample solution in a chromatographic column, and scanning the test sample solution by using an ultra-high performance liquid chromatograph to obtain a chromatogram of the test sample solution; and (4) identifying common peak components of a chromatogram by using reference substances including gallic acid, protocatechuic acid, protocatechuic aldehyde, procyanidine B2, epicatechin and rutin. And (5) establishing a fingerprint spectrum. According to the present invention, the precision of the used super effect liquid chromatograph is high, the repeatability of the test solution preparation method is good, the stability of the fingerprint established based on the two points is strong, the fingerprint establishment speed is fast, the sensitivity is high, and the chromatographic peak separation degree is good.

Description

Technical Field

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

[0002] The 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 usually includes thin layer chromatography, high performance liquid phase content determination method, fingerprint method, etc. As a single Chinese medicine preparation, Weimaining Capsules have relatively prescribed processing methods and usage methods, but due to the relatively complex chemical components it contains, the pharmacological effects have multi-target and multi-level characteristics, and there are many interfering factors, it is difficult to accurately control the quality and process of Weimaining Capsules from raw materials to products, and it is difficult to fully reflect the quality status of Weimaining Capsules products.

[0003] Fingerprint is an effective means of comprehensive analysis to identify the authenticity of Chinese herbal medicines or Chinese patent medicines, and to evaluate the consistency and stability of the quality of raw materials, semi-finished products and finished products using modern analytical technology. Compared with the existing identification of single active ingredients or index ingredients, the fingerprint technology of Chinese medicine integrates various effective chemical information to make the results more scientific and reasonable. Fingerprint of Chinese medicine has become an important method for quality control of Chinese medicine recognized at home and abroad. The research and establishment of fingerprint is of great significance to improving the quality of Chinese medicine and promoting the modernization of Chinese medicine. Ultra-high performance liquid chromatography (UPLC) has the advantages of strong sensitivity, high efficiency and good stability. It is currently a widely used method for fingerprint detection of Chinese medicine. It can separate multiple components in a short time with good separation. At present, there is no research on the establishment of fingerprint detection method and component identification of Weimaining by UPLC. Therefore, the study of using UPLC to establish fingerprint of Weimaining capsule is of great significance for quality control analysis of Weimaining capsule. Summary of the invention

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

[0005] (I) Preparation of test solution:

[0006] Take 2g of the contents of Weimaining capsules, place in a stoppered conical flask, add 50ml of water, heat and reflux for 20-40 minutes, cool, filter, concentrate the filtrate under reduced pressure, add 50ml of dilute ethanol, heat and reflux for 20-40 minutes, take out, cool, filter, take 25ml of the subsequent filtrate, concentrate under reduced pressure, wash the residue with acetonitrile-water mixed solution in batches, transfer the washing liquid to a 10ml volumetric flask, dilute to the scale with acetonitrile-water mixed solution, shake well, centrifuge, take 5ml of the supernatant, add to the 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 to a 10ml volumetric flask, dilute to 10ml with acetonitrile-water mixed solution, shake well, filter, and take the subsequent filtrate as the test solution;

[0007] (ii) respectively examine the precision of the ultra-high performance liquid chromatograph, 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] (iv) Use the reference substances gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin to identify the common peak components of the chromatogram;

[0010] (V) Establish fingerprint map.

[0011] Further, the fingerprint map is established through the following steps:

[0012] Take 15 batches of Weimaining capsules to prepare test solutions according to step (i), 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] Further, 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, the provided method for preparing the test solution has good repeatability, and the stability of the fingerprint spectrum established based on the above two points is strong. At the same time, compared with the method for establishing the HPLC fingerprint spectrum, the present invention establishes the fingerprint spectrum faster, with higher sensitivity and better separation of the chromatographic peaks. BRIEF DESCRIPTION OF THE DRAWINGS

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

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

[0028] Figure 3 This is a chromatogram of the test product 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 in the present invention;

[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 in the present invention;

[0031] Figure 6 This is a schematic diagram of the chromatographic peaks of the test sample and the reference sample when the extraction time is 40 minutes in the present invention;

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

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

[0034] Fig. 9 The reference fingerprints of 15 batches of test products of the present invention;

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

[0036] Fig.11 is a chromatogram of the mixed reference substance solution of the present invention;

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

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

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

[0040] Fig.15 The chromatogram of the test solution obtained by eluting with a C gradient in the present invention;

[0041] Fig.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] Fig.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] Fig.18 The chromatographic column of the present invention is Chromatogram of the test solution obtained at (1.7μm, 150*2.1mm);

[0044] Fig.19 The chromatogram of the test solution obtained when the flow rate of the present invention is 0.2 mL / min;

[0045] Fig. 20 This is the chromatogram of the test solution obtained when the flow rate of the present invention is 0.3 mL / min;

[0046] Fig.21 This is the chromatogram of the test solution obtained when the flow rate of the present invention is 0.4 mL / min;

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

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

[0049] Fig.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 the technical solution more clear, the technical solution is further described in detail below 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 the 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 Standard Technical Services Co., Ltd.), protocatechuic aldehyde (102752920, Shanghai McLean 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] (I) Preparation of test solution:

[0057] Take 2g of Weimaining capsules, place 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. Add acetonitrile-water (10:90) mixed solution to wash the residue in several times, transfer the washing solution to a 10ml volumetric flask, add acetonitrile-water (10:90) mixed solution to dilute to the scale, shake well, and centrifuge (speed is 30 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 it 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 the reference sample prepared by the heating reflux method and the ultrasonic dissolution method is shown, wherein 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 embodiment carried out 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 Figure 2, when heated to reflux for 20 minutes, the chromatographic peak area is 376297. Figure 5 As shown in Figure 1, when heated to reflux for 30 minutes, the chromatographic peak area is 1035856. Figure 6 As shown, when heated to reflux for 40 minutes, the chromatographic peak area is 492753. The results show that the peak area obtained by extracting 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, weigh accurately, place in a conical flask, add 50mL of 10% acetonitrile accurately, dissolve by ultrasonication, let stand for 1h, heat under reflux for 1h, cool, weigh again, make up the lost amount with 10% acetonitrile, shake well, and take the filtrate as the test solution. Figure 2 Shown is a chromatogram of the test product obtained by sonication with 10% acetonitrile.

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

[0064] Also, see Figure 1 The chromatogram of the test product 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 separation of most peaks. This proves the effectiveness of the method for preparing the sample solution disclosed in the present invention.

[0065] (ii) The precision of the ultra-high performance liquid chromatography, 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, weigh it accurately, place it in a 10mL volumetric flask, add 50% methanol and ultrasonically dissolve and dilute to the scale to make a reference substance solution containing 1 mg epicatechin per 1mL.

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

[0069] The same test solution was sampled 6 times in succession. Taking epicatechin as the reference peak (S), the relative retention time and relative standard deviation (RSD) of the other common peaks were calculated. The results showed that the RSD of the relative retention time of each common peak was <1% (Table 1), and the RSD of the relative peak area was <2.5% (Table 2), indicating that the instrument had good 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 (I) above. The samples were injected separately, and epicatechin was used as the reference peak (S). The RSDs of the relative retention time and relative peak area of ​​the remaining common peaks were calculated. The results showed that the RSDs of the relative retention time of the common peaks were <2% (Table 3), and the RSDs of the relative peak areas were <5% (Table 4), indicating that the repeatability of the test solution preparation method was good.

[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 taken and sampled 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] (iii) 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.

[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 conducts comparative experiments on different types of chromatographic columns, and obtains the following results: Figure 16-18 The chromatographic columns shown are T3 (1.6μm, 2.1*150mm), HSS T3 (1.8μm, 2.1*150mm) and The chromatogram of the test solution obtained when the column diameter is 1.7 μm, 150*2.1 mm. By comparing the baseline stability and separation effect of the chromatographic peak, the chromatographic column model is preferably T3 (1.6μm, 2.1*150mm).

[0090] Secondly, the embodiments of the present invention are 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 are shown. 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 a comparative experiment 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 reference substances such as 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 make up the volume. Dissolve the remaining reference substances in 50% methanol, make up the volume, and shake well. Take 200 μL of each of the above reference substance solutions, mix well, and use as 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 by PDA detector in the range of 210-400 nm 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] (V) Establish fingerprint map.

[0110] Take 15 batches of Weimaining capsules to prepare test solutions according to step (i), 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 taken 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, and the sample S1 spectrum was set as the reference spectrum. The median method was used to generate the reference spectrum, with a time window width of 0.1, and automatic matching was used to generate a reference fingerprint spectrum.

[0114] Furthermore, the generated reference fingerprint was used to evaluate the stability of the quality of the Weimaining capsule samples. Specifically, the similarity between the UPLC spectrum of each batch and its reference fingerprint was calculated. Figure 9-10 The reference 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 reference 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 contents are only preferred embodiments of the present invention. For ordinary technicians in this field, many changes can be made in the specific implementation methods and application scopes 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 in a stoppered conical flask, add 50ml of water, heat and reflux for 20-40 minutes, cool, filter, concentrate the filtrate under reduced pressure, add 50ml of dilute ethanol, heat and reflux for 20-40 minutes, take out, cool, filter, take 25ml of the subsequent filtrate, concentrate under reduced pressure, wash the residue with acetonitrile-water mixed solution in batches, transfer the washing liquid to a 10ml volumetric flask, dilute to the scale with acetonitrile-water mixed solution, shake well, centrifuge, take 5ml of the supernatant, add to the 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 to a 10ml volumetric flask, dilute to 10ml with acetonitrile-water mixed solution, shake well, filter, and take the subsequent filtrate as the test solution; (ii) respectively examine the precision of the ultra-high performance liquid chromatograph, the repeatability of the test solution preparation method and the stability of the test solution; (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; (iv) Use the reference substances gallic acid, protocatechuic acid, protocatechuic aldehyde, proanthocyanidin B2, epicatechin, and rutin to identify the common peak components of the chromatogram; (V) Establish fingerprint map.

2. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: The fingerprint is created by the following steps: Take 15 batches of Weimaining capsules to prepare test solutions according to step (i), 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; 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 column 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 rpm and the centrifugal time was 5 min.

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. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 1, characterized in that: 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; the injection volume is: 3 μL.

8. The method for establishing the UPLC fingerprint of Weimaining Capsules according to claim 2, characterized in that: The gradient elution values ​​are: timemin(0, 10, 17, 25, 30, 35, 40); Acetonitrile (7%, 8%, 10%, 12%, 12%, 14%, 14%); 0.1% phosphoric acid (93%, 92%, 90%, 88%, 88%, 86%, 86%).

9. A UPLC fingerprint of Weimaining Capsules, characterized by: Derived from the establishment method according to claims 1-8.

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