Establishment method of fingerprint spectrum of bock greenbrier rhizome preparation as well as fingerprint spectrum and application of fingerprint spectrum

Abstract

The invention provides a method for establishing a fingerprint spectrum of a bock greenbrier rhizome preparation, which comprises the following steps: S1, preparation of a test solution: weighing the bock greenbrier rhizome preparation, adding methanol, carrying out ultrasonic treatment, and filtering to obtain the test solution; s2, preparation of a reference substance solution: weighing eight reference substances including cryptochlorogenic acid, neochlorogenic acid, chlorogenic acid, polydatin, astilbin, oxyresveratrol, quercitrin and resveratrol, mixing, and adding methanol to prepare the reference substance solution; and S3, establishment of a fingerprint: respectively measuring the prepared test solution and the reference solution by using HPLC, recording and comparing chromatograms, and identifying common peaks to obtain the fingerprint of the bock greenbrier rhizome preparation. According to the method, the fingerprint spectrum of the bock greenbrier rhizome preparation is successfully established by optimizing the extraction conditions of the bock greenbrier rhizome preparation and the HPLC chromatographic conditions, the establishment method of the fingerprint spectrum is accurate, reliable and simple to operate, and the detection method is high in precision and good in specificity, stability and repeatability.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine fingerprint analysis, and particularly relates to a method for establishing a fingerprint of a gypsophila strychni preparation and a fingerprint thereof. Background Art

[0002] Jingangteng preparations are single-ingredient preparations made from the Jingangteng plant of the Liliaceae family through water extraction and alcohol precipitation. They have the effects of clearing away heat and detoxifying, reducing swelling and dispersing nodules. The main ones include Jingangteng syrup and Jingangteng capsules.

[0003] At present, the quality control of Jingangteng Syrup is still mainly based on ministerial standards, mainly using TLC comparative analysis of raw medicinal materials and foam experiments for qualitative identification. The method is too simple and difficult to achieve the purpose of quality control. The existing statutory quality standards for Jingangteng Capsules are included in the "National Drug Standard New Drug Standard Volume 35", and its content determination item uses thin-layer scanning to determine diosgenin. This method has poor reproducibility, and the ingredients of traditional Chinese medicine are complex. Its pharmacological effects are often the result of the joint action of multiple components. The content determination of a single component cannot comprehensively and scientifically evaluate the quality of traditional Chinese medicine.

[0004] Fingerprint is an important method for evaluating the batch uniformity of Chinese patent medicine preparations. At the same time, multi-component content determination is the main content determination method advocated in recent years. The combination of the two is important for effectively controlling the batch uniformity and effectiveness of preparations. Based on this, it is necessary to establish a method for simultaneously determining the content of the main components of the Jingangteng preparation fingerprint. Summary of the invention

[0005] The purpose of the present invention is to provide a method for establishing a fingerprint of a gypsophila preparation, which can at least solve some of the defects in the prior art.

[0006] To achieve the above object, the present invention adopts the following technical solution:

[0007] A method for establishing a fingerprint spectrum of a gypsophila preparation comprises the following steps:

[0008] S1. Preparation of the test solution: weigh the Kanthus chinensis preparation, add methanol, perform ultrasonic treatment, and filter to obtain the test solution;

[0009] S2. Preparation of reference solution: weigh 8 reference substances including cryptochlorogenic acid, neochlorogenic acid, chlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol, mix them, add methanol to prepare a reference solution;

[0010] S3. Establishment of fingerprint: The test solution and reference solution prepared as above are measured by HPLC (high performance liquid chromatography), the chromatograms are recorded and compared, the common peaks are identified, and the fingerprint of the Kantung vine preparation is obtained.

[0011] Furthermore, in step S1, the methanol concentration is 90%, the ultrasonic treatment power is 200 W, the frequency is 40 kHz, and the ultrasonic treatment time is 15 min.

[0012] Furthermore, in the reference solution, the concentration of cryptochlorogenic acid was 2.190 μg / mL, the concentration of neochlorogenic acid was 18.914 μg / mL, the concentration of chlorogenic acid was 46.320 μg / mL, the concentration of polydatin was 1.661 μg / mL, the concentration of astilbin was 46.379 μg / mL, the concentration of oxidized resveratrol was 12.524 μg / mL, the concentration of quercetin was 46.411 μg / mL, and the concentration of resveratrol was 2.5967 μg / mL.

[0013] Furthermore, in step S3, the chromatographic conditions of HPLC are as follows: the chromatographic column is CAPCELL PAK C18, 250 mm×4.6 mm, 5 μm; the mobile phase is 0.1% phosphoric acid solution-acetonitrile, and gradient elution is performed; the detection wavelength is 200 nm to 400 nm; the flow rate is 1 mL / min; the column temperature is 25°C to 40°C; the injection volume is 10 μL; and the number of theoretical plates calculated based on the chlorogenic acid peak should be no less than 10,000.

[0014] Furthermore, the chromatographic conditions of the HPLC are as follows: the chromatographic column is CAPCELLPAK C18, 250 mm×4.6 mm, 5 μm; the mobile phase is 0.1% phosphoric acid solution-acetonitrile, with gradient elution; the detection wavelength is 303 nm; the flow rate is 1 mL / min; the column temperature is 35°C; the injection volume is 10 μL; and the theoretical plate number calculated based on the chlorogenic acid peak should be no less than 10,000.

[0015] Further, the gradient elution program is as follows by volume percentage: 0-5 min, 6% acetonitrile, 94% 0.1% phosphoric acid solution; 5-15 min, 6%-10% acetonitrile, 94%-90% 0.1% phosphoric acid solution; 15-60 min, 10%-29% acetonitrile, 90%-71% 0.1% phosphoric acid solution; 60-65 min, 29%-6% acetonitrile, 71%-94% 0.1% phosphoric acid solution; 65-70 min, 6% acetonitrile, 94% 0.1% phosphoric acid solution.

[0016] Furthermore, in step S3, the chromatogram of the test solution is imported into the "Chinese medicine chromatographic fingerprint similarity evaluation system software" to perform similarity evaluation with the chromatogram of the reference solution to determine the similarity and identify the common peaks.

[0017] Furthermore, the gypsophila paniculata preparation is gypsophila paniculata syrup or gypsophila paniculata capsule.

[0018] In addition, the present invention also provides a fingerprint spectrum obtained by the method for establishing the fingerprint spectrum of the above-mentioned smilax china preparation.

[0019] The present invention also provides the application of the method for establishing the fingerprint of the above-mentioned strychnos nux vomica preparation in the determination of the content of the ingredients of the strychnos nux vomica preparation and the quality inspection of the strychnos nux vomica preparation.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) The present invention successfully established the fingerprint of the Kungfu Herb preparation by optimizing the extraction conditions and HPLC chromatography conditions of the Kungfu Herb preparation. The fingerprint establishment method is accurate, reliable, and easy to operate. The detection method has high precision, specificity, stability, and good repeatability.

[0022] (2) The present invention can simultaneously detect the contents of eight active ingredients in the Knotweed preparation, including neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin, and resveratrol, providing a basis for the quality control of the Knotweed preparation.

[0023] The present invention will be further described in detail below with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 15 batches of HPLC superimposed graphs of Jingangteng syrup and the schematic diagram of reference graph R in Example 1;

[0025] Figure 2 is the HPLC chromatogram of the reference substance solution in Example 1;

[0026] Figure 3 is the HPLC chromatogram of the jingangteng syrup in Example 1;

[0027] Figure 4 is the HPLC chromatogram of the blank sample solution in Example 1;

[0028] Figure 5 HPLC chromatograms of the Jingangteng syrup in Example 2 under different extraction solvents; wherein (a) methanol, (b) 90% methanol, and (c) ethanol;

[0029] Figure 6 HPLC chromatograms of different extraction methods of the jingangteng syrup in Example 2; wherein (a) ultrasonic extraction, (b) reflux extraction;

[0030] Figure 7 The HPLC chromatograms of the Jingangteng syrup in Example 2 at different extraction times, wherein (a) 15 min, (b) 30 min, and (c) 60 min;

[0031] Figure 8The HPLC chromatograms of the jingangteng syrup in Example 2 using different mobile phases; wherein (a) 0.1% phosphoric acid (A)-acetonitrile (B), (b) 0.1% glacial acetic acid (A)-acetonitrile (B), (c) 0.1% formic acid (A)-acetonitrile (B);

[0032] Fig. 9 HPLC chromatograms of the Jingangteng syrup in Example 2 at different detection wavelengths; (a) 203 nm, (b) 290 nm, (c) 303 nm;

[0033] Fig.10 The HPLC chromatograms of the jingangteng syrup in Example 2 at different column temperatures are shown in Figure 2; wherein (a) is 25°C, (b) is 30°C, (c) is 35°C, and (d) is 40°C;

[0034] Fig.11 The HPLC chromatograms of the Jingangteng syrup in Example 2 using different chromatographic columns; (a) WondaSil C18, (b) Nano Chrom C18, (c) SHIMADZU C18, (d) Thermo scientific C18, (e) Agilent 5TC-C18 (2) C18, (f) GL Sciences Inc Inertsil ODS-3 C18, (g) Capcellpak C18, (f) Waters C18;

[0035] Fig.12 The HPLC chromatograms of the Jingangteng syrup in Example 2 at different flow rates, wherein (a) 0.8 mL / min, (b) 1.0 mL / min, and (c) 1.2 mL / min;

[0036] Fig.13 The HPLC chromatograms of the Jingangteng syrup in Example 2 at different running times are shown in (a) 65 min, (b) 70 min, and (c) 75 min;

[0037] Fig.14 Schematic diagram of HPLC superimposed patterns and reference pattern R of 15 batches of Jingangteng Capsules in Example 4;

[0038] Fig.15 is the HPLC chromatogram of the reference substance solution in Example 4;

[0039] Fig.16 is the HPLC chromatogram of Jingangteng Capsule in Example 4;

[0040] Fig.17 is the HPLC chromatogram of the blank sample solution in Example 4;

[0041] Fig.18 HPLC chromatograms of Jingangteng capsule in Example 4 under different extraction solvents; wherein (a) methanol, (b) 90% methanol, and (c) 60% methanol;

[0042] Fig.19 HPLC chromatograms of Jingangteng Capsules in Example 4 using different extraction methods; (a) ultrasonic extraction, (b) reflux extraction;

[0043] Fig. 20 HPLC chromatograms of Jingangteng capsules at different extraction times in Example 4; (a) 15 min, (b) 30 min, (c) 60 min;

[0044] Fig.21 The HPLC chromatograms of Jingangteng Capsules in Example 4 using different mobile phases; wherein (a) 0.1% phosphoric acid (A)-acetonitrile (B), (b) 0.1% glacial acetic acid (A)-acetonitrile (B), (c) 0.1% formic acid (A)-acetonitrile (B);

[0045] Fig. 22 HPLC chromatograms of Jingangteng Capsules in Example 4 at different detection wavelengths; (a) 280 nm, (b) 303 nm, (c) 326 nm;

[0046] Fig.23 The HPLC chromatograms of Jingangteng Capsules in Example 4 at different column temperatures are shown in Figure 4; (a) 25°C, (b) 30°C, and (c) 35°C;

[0047] Fig.24 The HPLC chromatograms of Jingangteng Capsules in Example 4 using different chromatographic columns; (a) CAPCELLPAK C18, (b) GL Science WondaSil C18, (c) NanoChrom ChromCoreC18, and (d) WatersSunFire C18. DETAILED DESCRIPTION

[0048] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0049] Experimental instruments and reagents:

[0050] 1. Instrument

[0051] Electronic balance [Mettler Toledo MS105, Mettler Toledo Instrument (Shanghai) Co., Ltd.]; ultrasonic cleaner (KQ-500DE, Kunshan Ultrasonic Instrument Co., Ltd.); ultrapure water machine (PGJ-20-YS, Wuhan Pinguan Instrument Equipment Co., Ltd.); electric constant temperature water bath (DK-900-II, Tianjin Test Instrument Co., Ltd.); high performance liquid chromatograph [UltiMate3000, Thermo Fisher Scientific (China) Co., Ltd.], including DAD detector (DAD-300) and chromatography workstation (Chromelenon 7).

[0052] 2. Drug test reagents

[0053] Cryptochlorogenic acid (purity 98.0%, CAS No. 905-99-7); neochlorogenic acid (purity 98.0%, CAS No. 906-33-2); oxidized resveratrol (purity 98.0%, CAS No. 29700-22-9) were purchased from Shanghai Topu Biopharmaceutical Technology Co., Ltd.; polydatin (purity 87.3%, batch No. 111575-201603), astilbin (purity 95.0%, batch No. 111798-202306), quercetin (purity 95.3%, batch number 111538-202308), chlorogenic acid [purity 96.3%, batch number 110753-202119], quercetin (purity 99.1%, batch number 100081-201610), and resveratrol (purity 99.4%, batch number 111535-201703) were purchased from China Food and Drug Inspection Institutes.

[0054] There are 15 batches of Kyoritsa syrup (numbered S1 to S15), with batch numbers 20220101, 20220605, 20220901, 20220902, 20220903, 20220904, 20221101, 20230101, 20230102, 20240101, 20240102, 20240103, 20240104, 20240105, and 20240106, all provided by Hubei Furen Pharmaceutical Co., Ltd.

[0055] There are 15 batches of Jingangteng Capsules (numbered S1 to S15), with batch numbers 20210101, 20210103, 20210105, 20210108, 20220402, 20231217, 20231218, 20231219, 20231220, 20240105, 20240106, 20240107, 20240108, 20240109, and 20240110, all provided by Hubei Furen Pharmaceutical Co., Ltd.

[0056] Phosphoric acid (superior grade, Tianjin Komiou Chemical Reagent Co., Ltd.), water is ultrapure water prepared by Pinguan ultrapure water machine, acetonitrile and methanol (chromatographic grade, Shanghai Sarao Industrial Co., Ltd.).

[0057] Example 1: Establishment of fingerprint of jingangteng syrup

[0058] 1. Solution preparation

[0059] 1.1 Reference solution: Accurately weigh appropriate amounts of 8 reference substances including cryptochlorogenic acid, neochlorogenic acid, chlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol, add 90% methanol to prepare a mixed solution containing 2.190 μg / mL of cryptochlorogenic acid, 18.914 μg / mL of neochlorogenic acid, 46.320 μg / mL of chlorogenic acid, 1.661 μg / mL of polydatin, 46.379 μg / mL of astilbin, 12.524 μg / mL of oxidized resveratrol, 46.411 μg / mL of quercetin and 2.597 μg / mL of resveratrol.

[0060] 1.2 Test solution: Accurately take 10 mL of Vinegar syrup, place it in a 50 mL volumetric flask, add appropriate amount of 90% methanol, treat it with ultrasound (200W, 40kHZ) for 15 minutes, take it out, let it stand to room temperature, make up to the volume with 90% methanol, shake well, pass it through a microporous filter membrane (0.22μm), and take the filtrate to obtain the solution.

[0061] 2. Chromatographic conditions

[0062] The chromatographic column was (Capcell Pak C18, 4.6 mm × 250 mm, 5.0 μm), the mobile phase was 0.1% phosphoric acid solution (A)-acetonitrile (B), and the elution was performed according to the gradient elution program shown in Table 1, the column temperature was 35°C, the flow rate was 1.0 mL / min, the detection wavelength was 303 nm, and the injection volume was 10 μL.

[0063] Table 1: Gradient elution program

[0064]

[0065] 3. Investigation of fingerprint methodology

[0066] 3.1 Precision test: Take the syrup of golden vine (S10), prepare the test solution according to the method of "1.2 Test solution", measure 6 times according to "2, chromatographic conditions", record the peak area of ​​each common peak, calculate the peak area RSD of 8 components of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol, which are 0.55%, 0.52%, 0.55%, 1.11%, 0.88%, 0.55%, 1.42% and 0.77% respectively. It can be seen that the relative peak area RSD of each common peak is <1.5%, indicating that the precision of the instrument is good.

[0067] 3.2 Stability test: Take the syrup of golden vine (S10), prepare the test solution according to the method of "1.2 Test solution", measure according to "2, chromatographic conditions", record the relative peak area of ​​each common peak within 0, 4, 8, 12, 16, and 24 hours, and calculate the RSD of the peak area of ​​8 components of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol within 24 hours, which are 0.68%, 0.43%, 0.70%, 0.94%, 0.43%, 0.83%, 1.80%, and 0.99%, respectively. It can be seen that the relative peak area RSD of each common peak is <2.0%, indicating that the stability of the golden vine syrup is good within 24 hours.

[0068] 3.3 Repeatability test: Take the syrup of golden vine (S10), prepare 6 test solutions according to the method of "1.2 Test solution", measure according to "2, chromatographic conditions", record the peak area of ​​each common peak, calculate the RSD of the peak area of ​​8 components of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol, which are 0.55%, 0.10%, 0.18%, 0.74%, 0.20%, 0.72%, 0.52% and 0.70% respectively. It can be seen that the relative peak area RSD of each common peak is <1.0%, indicating that the method has good repeatability.

[0069] 3.4 Establishment of fingerprints and identification of chromatographic peaks: Take 15 batches of Vinegar syrup, prepare the test solution according to the method of "1.2 Test solution", inject according to the chromatographic conditions under "2. Chromatographic conditions", record the chromatograms, set the integration parameters on the Dion Chromeleon chromatography workstation (version 7.2.10), and integrate the main chromatographic peaks. Export the CDF files of the spectra one by one in the form of CDF data files. Import the exported CDF files of the above 15 batches of chromatograms into the "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition)", cut off the solvent peaks in the first 3 minutes, and establish the overlay diagrams and control fingerprints of 15 batches of samples through the evaluation system, as shown in the figure. Figure 1 As shown. By comparing with the chromatogram of the mixed reference solution (such as Figure 2As shown) comparison, Figure 1 Eight common peaks were identified, in order: peak 3 neochlorogenic acid, peak 5 chlorogenic acid, peak 6 cryptochlorogenic acid, peak 12 polydatin, peak 14 astilbin, peak 16 oxidized resveratrol, peak 18 quercetin, peak 21 resveratrol; thus, the fingerprint of the golden vine syrup was obtained as follows Figure 3 As shown, the eight common peaks were renumbered from 1 to 8, and peaks 1 to 8 represented neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin, and resveratrol, respectively.

[0070] 3.5 Similarity evaluation: The similarity between the chromatograms of 15 batches of Jingangteng syrup and the control fingerprint was calculated using the "Chinese Herbal Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition)", and the similarity results are shown in Table 2. As can be seen from Table 2, the similarities are all > 0.90, indicating that different batches of Jingangteng syrup have high uniformity and stability in chemical composition, similar content of main components, no significant quality differences, and their fingerprint characteristics are effectively and accurately reflected in the fingerprint.

[0071] Table 2: Similarity calculation results of 15 batches of jingangteng syrup

[0072]

[0073] 4. Blank sample test

[0074] Take the auxiliary materials of Jingangteng Syrup to prepare a negative blank solution, inject 10 μL of it using the chromatographic conditions under "2. Chromatographic conditions", and record the chromatogram, such as Figure 4 The results show that the negative blank solution does not have a chromatographic peak corresponding to the test solution in the chromatogram, and there will be no interference.

[0075] Example 2: Preparation method of Jingangteng syrup test solution and optimization of chromatographic conditions

[0076] 1. Selection of extraction solvent

[0077] Take three portions of King Kong Teng Syrup S10 (batch number: 20240101), accurately add 50 mL of methanol, 90% methanol solution and ethanol solution respectively, weigh the weight, ultrasonically treat at a power of 200 W and a frequency of 40 kHz for 15 minutes, cool, weigh again, make up the lost weight with the corresponding solvent, shake well, filter, and take the filtrate as the test solution. Determine under the same method, the chromatograms are as follows Figure 5 (a) Figure 5 (b) Figure 5(c) shows that the best extraction solvent was determined by comparison. The results showed that the chromatographic peaks of ethanol extraction were few and small, and the extraction was incomplete, while the main chromatographic peaks of 90% methanol extraction were higher than those of methanol extraction, with high extraction efficiency and better peak shape; therefore, 90% methanol was selected as the extraction solvent for the preparation of the test sample of the golden vine syrup.

[0078] 2. Selection of extraction method

[0079] Take two portions of King Kong Teng Syrup S10 (batch number: 20240101), accurately add 50 mL of 90% methanol solution, weigh the weight, treat with ultrasound (power 200W, frequency 40kHz) for 15 minutes and reflux extraction for 15 minutes, cool, weigh again, make up the lost weight with 90% methanol, shake well, filter, and take the filtrate as the test solution. Determine under the same method, the chromatograms are as follows Figure 6 (a) and Figure 6 (b) shows that the best extraction method was determined by comparison. The results showed that the main chromatographic peak height in the chromatogram was higher and the peak shape was better when ultrasonic extraction was used; therefore, ultrasonic treatment was selected as the extraction method for preparing the sample of the syrup of the golden vine.

[0080] 3. Selection of extraction time

[0081] Take three portions of King Kong Teng Syrup S10 (batch number: 20240101), accurately add 50 mL of 90% methanol solution, weigh the weight, and treat with ultrasound (power 200W, frequency 40kHz) for 15min, 30min, and 60min respectively, cool, weigh again, make up the lost weight with 90% methanol, shake well, filter, and take the filtrate as the test solution. Determine under the same method, the chromatograms are as follows Figure 7 (a) Figure 7 (b) and Figure 7 (c) shows that the optimal extraction time was determined by comparison. The results showed that there was no significant difference in the chromatograms of ultrasonic extraction for 15 min and 30 min, and the peak shape was good. However, after ultrasonic treatment for 60 min, the peak areas of multiple chromatographic peaks became smaller. In order to ensure good extraction efficiency and prevent the degradation of chemical components during the extraction process; therefore, ultrasonic treatment for 15 min was selected as the optimal extraction time for preparing the test sample of Jingangteng syrup.

[0082] 4. Selection of mobile phase

[0083] Take an appropriate amount of Jingangteng syrup S10 (batch number: 20240101), prepare it according to the test solution preparation method in Example 1, and compare the following mobile phases with a C18 chromatographic column: ① 0.1% phosphoric acid (A)-acetonitrile (B), ② 0.1% glacial acetic acid (A)-acetonitrile (B), ③ 0.1% formic acid (A)-acetonitrile (B), and perform gradient elution using the gradient elution program in Example 1. The chromatograms are as follows: Figure 8 (a) Figure 8 (b) Figure 8 As shown in (c), although the above three mobile phases all achieved separation, mobile phase ① had the best separation effect, while mobile phase ② had poor separation effects on peaks 2 and 3; when separated by mobile phase ③, the responses of most peaks were lower than those of mobile phase ①, and peaks 7 and 8 could not be completely separated; therefore, mobile phase ① was selected as the mobile phase for the test sample of Vinegar Thunb.

[0084] 5. Selection of detection wavelength

[0085] Take an appropriate amount of Jingangteng Syrup S10 (batch number: 20240101), prepare it according to the test solution preparation method of Example 1, inject it into the high performance liquid chromatograph for analysis, and follow the chromatographic conditions of Example 1, wherein the detection wavelengths are compared at 203nm, 290nm, and 303nm, respectively. The chromatograms are as follows: Fig. 9 (a) Fig. 9 (b) Fig. 9 (c) The results show that when the detection wavelength is 303nm, the chromatographic information is richer, the baseline is relatively stable, the response values ​​of each peak are relatively balanced, and it can relatively comprehensively reflect the component information in the sample, especially the peak areas of peaks 2 and 6 with lower responses are larger than those at other wavelengths; therefore, 303nm was selected as the detection wavelength for the Jingangteng syrup sample.

[0086] 6. Selection of column temperature

[0087] Take 4 portions of Jingangteng syrup S10 (batch number: 20240101), prepare according to the test solution preparation method of Example 1, inject into high performance liquid chromatography for analysis, wherein the column temperature is 25 ℃, 30 ℃, 35 ℃, 40 ℃, respectively, and the chromatograms are as follows: Fig.10 (a) Fig.10 (b) Fig.10 (c) Fig.10 (d) As shown. The results show that when the column temperature is 30℃, the chromatographic peak separation effect is not good, especially peaks 4 and 5 at 30℃, which include nearby impurity peaks, affecting the accuracy of the results; when the column temperature is 25℃, peak 8 may include impurity peaks, and when the column temperature is 35℃, the HPLC chromatogram separation effect of Jingangteng Syrup is the best; therefore, 35℃ is selected as the column temperature of Jingangteng Syrup fingerprint.

[0088] 7. Selection of chromatographic columns

[0089] Take an appropriate amount of Jingangteng Syrup S10 (batch number: 20240101), prepare it according to the test solution preparation method in Example 1, inject it into a high performance liquid chromatograph for analysis, and analyze it with chromatographic columns WondaSil C18, Nano Chrom C18, SHIMADZU C18, Thermo scientific C18, Agilent 5TC-C18 (2) C18, GL Sciences Inc Inertsil ODS-3C18, Capcell pak C18 and Waters C18, respectively. The chromatograms are as follows: Fig.11 (a) Fig.11 (b) Fig.11 (c) Fig.11 (d), 11(e), Fig.11 (f) Fig.11 (g) Fig.11 (h) As shown. The results show that when using WondaSil C18 column, peak 2 and peak 3 cannot be completely separated, and peak 6 cannot be separated from impurities; when using Nano Chrom C18 column, peak 5 cannot be separated from impurities, and all peaks have obvious tailing; when using SHIMADZU C18 column, peak 7 and peak 8 are not separated from impurities, and the impurity peak on the left side of peak 2 disappears; when using Thermo scientific C18 column, peak 7 and peak 8 are not separated from impurities; when using Agilent 5TC-C18(2)C18, peak 7 and peak 8 are not separated from impurities, and the impurity peak on the left side of peak 2 disappears; when using GL Sciences Inc Inertsil ODS-3C18 column, peak 6 is not separated from impurities, and the impurity peak on the left side of peak 2 disappears; when using Waters C18 column, the chromatographic peak tails seriously; therefore, Capcellpak C18 column was selected as the chromatographic column for the determination of the fingerprint of Jingangteng syrup.

[0090] 8. Selection of column flow rate

[0091] Take an appropriate amount of Jingangteng syrup S10 (batch number: 20240101), prepare it according to the test solution preparation method of Example 1, and inject it into a high performance liquid chromatograph for analysis, wherein the flow rates are 0.8mL / min, 1.0mL / min, and 1.2mL / min, respectively. The chromatograms are as follows: Fig.12 (a) Fig.12 (b) Fig.12(c) As shown. The results show that when the flow rate is 0.8 mL / min, peak 2 cannot be separated from the impurity peak on the left; when the flow rate is 1.2 mL / min, peak 6 appears as a double peak; and the flow rate of 1.0 mL / min shows advantages in terms of better separation, peak shape and system pressure control in the HPLC analysis of Jingangteng Syrup S10. Therefore, 1.0 mL / min was selected as the optimal flow rate.

[0092] 9. Selection of column running time

[0093] Take an appropriate amount of Jingangteng syrup S10 (batch number: 20240101), prepare it according to the test solution preparation method of Example 1, inject it into the high performance liquid chromatography for analysis, wherein the running time is 65min, 70min, and 75min respectively, and the chromatograms are as follows: Fig.13 (a) Fig.13 (b) Fig.13 (c) The results show that when the running time is 65 minutes, peak 8 may contain impurities; when the running time is 75 minutes, peak 6 has double peaks; and when the running time is 70 minutes, each chromatographic peak is clear and well separated, so 70 minutes is selected as the best running time.

[0094] Example 3: Determination of the contents of 8 ingredients in Jingangteng syrup

[0095] 1. Preparation of solution: same as in Example 1 above.

[0096] 2. Chromatographic conditions: same as in Example 1 above.

[0097] 3. Determination of ingredient content and methodology investigation

[0098] 3.1 Linear relationship investigation: After each reference substance was accurately weighed, it was dissolved in 90% methanol to prepare a mixed solution containing 1623.860μg / mL of cryptochlorogenic acid, 840.840μg / mL of neochlorogenic acid, 2010.744μg / mL of chlorogenic acid, 521.181μg / mL of polydatin, 951.900μg / mL of astilbin, 530.180μg / mL of oxidized resveratrol, 428.850μg / mL of quercetin, and 381.828μg / mL of resveratrol. After stepwise dilution, the chromatogram was recorded according to "2. Chromatographic conditions", and the horizontal axis (x, μg / mL) represented the mass concentration, and the vertical axis (y) represented the peak area. The linear analysis of each component was performed, and the results are shown in Table 3. It can be seen from Table 3 that r is greater than 0.9999, and each component has a good linear relationship within its respective range.

[0099] Table 3: Linear equations and linear ranges of 8 components

[0100]

[0101] 3.2 Precision test: Same as “3.1 Precision test” in Example 1 above.

[0102] 3.3 Stability test: Same as “3.2 Stability test” in Example 1 above.

[0103] 3.4 Repeatability test: Same as “3.3 Repeatability test” in above Example 1.

[0104] 3.5 Sample recovery test: Take 6 portions of the same Jingangteng syrup sample (S10), accurately measure 5 ml of each portion, and then add nearly equal amounts of each single reference substance, prepare the test solution according to the method of "1.2 Test solution", and determine according to "2, chromatographic conditions", and calculate the average sample recovery of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol. The results are shown in Table 4. The results show that the average sample recovery of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol are 102.3%, 96.5%, 97.4%, 103.4%, 98.7%, 95.6%, 108.4% and 100.6% (n=6), respectively, indicating that the recovery rate of this method is good.

[0105] Table 4: Recovery rates of various components in syrup of golden vine

[0106]

[0107]

[0108] 3.6 Sample content determination: Take the Vinegar syrup (S1~S15), prepare the test solution according to the method of "1.2 Test solution", and perform sample injection and determination according to "2. Chromatographic conditions". Calculate the contents of the 8 components in the sample. The results are shown in Table 5.

[0109] Table 5: Determination results of 8 components in 15 batches of Jingangteng syrup (μg / mL)

[0110]

[0111] Example 4: Establishment of fingerprint of Jingangteng capsule

[0112] 1. Solution preparation

[0113] 1.1 Reference solution: same as in Example 1 above.

[0114] 1.2 Test solution: Same as the above Example 1, except that the test sample is Rhizoma Cyperi capsule.

[0115] 2. Chromatographic conditions: same as in Example 1 above.

[0116] 3. Investigation of fingerprint methodology

[0117] 3.1 Precision test: Take the test solution of Jingangteng Capsule (S10), and inject it continuously for 6 times according to the chromatographic conditions under "2. Chromatographic conditions", and record the retention time and peak area of ​​each common peak. Among them, the peak area RSD values ​​of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol are 1.25%, 0.89%, 0.92%, 0.98%, 1.07%, 1.01%, 1.32% and 1.23% respectively, and the peak area RSD of each component is <1.26%, and the calculated retention time RSD of each common peak is <0.12%, indicating that the instrument has good precision.

[0118] 3.2 Stability test: Take Jingangteng Capsule (S10), prepare the test solution according to the method under "1.2 Test solution", and inject and measure at 0, 4, 8, 12, 16, 20, and 24 hours according to the chromatographic conditions under "2. Chromatographic conditions", and the peak area results of each component at 7 time points are shown in Table 6. As shown in Table 6, the RSD of the retention time of each common peak is <0.06%, and the RSD of the peak area is <1.85%, indicating that the test solution has good stability within 24 hours.

[0119] Table 6: Peak area results of stability test

[0120]

[0121]

[0122] 3.3 Repeatability test: Take the same Jingangteng capsule sample (S10), prepare 6 test solutions according to the method under "1.2 Test solution", inject and measure under the chromatographic conditions under "2. Chromatographic conditions", calculate the retention time RSD value and peak area RSD value of each component, and the results are shown in Table 7. As can be seen from Table 7, the peak area RSD of each common peak is <2.43%, and the retention time RSD is <0.14%, indicating that the method has good repeatability.

[0123] Table 7: Repeatability test peak area investigation results

[0124]

[0125] 3.4 Establishment of fingerprints and identification of chromatographic peaks: Take 15 batches of Jingangteng capsules, prepare the test solution according to the method under "1.2 Test solution", inject according to the chromatographic conditions under "2. Chromatographic conditions", record the chromatogram, and integrate the main chromatographic peaks. Export the CDF files of the spectra one by one in the form of CDF data files, and import the exported CDF files of the above 15 batches of chromatograms into the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition)". Through the evaluation system, 15 batches of sample overlays and reference fingerprints were established, as shown in the following figure. Fig.14 A total of 8 common peaks were identified and compared with the chromatogram of the mixed reference substance (such as Fig.15 ) were compared, and the eight common peaks were identified and re-numbered in order: peak 1 is neochlorogenic acid, peak 2 is chlorogenic acid, peak 3 is cryptochlorogenic acid, peak 4 is polydatin, peak 5 is astilbin, peak 6 is oxidized resveratrol, peak 7 is quercetin, and peak 8 is resveratrol. Fig.16 shown.

[0126] 3.5 Similarity evaluation: The similarity between the chromatograms of 15 batches of Jingangteng capsules and the control fingerprint was calculated using the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System Software (2012 Edition)", and the similarity results are shown in Table 8. As can be seen from Table 8, the similarity is between 0.860 and 0.991, indicating that the uniformity between Jingangteng capsules is good.

[0127] Table 8: Similarity calculation results of 15 batches of Jingangteng capsules

[0128]

[0129] 4. Blank sample test

[0130] Take the excipients of Jingangteng Capsule to prepare a negative blank solution, inject 10 μL of the solution using the chromatographic conditions under “2. Chromatographic conditions”, and record the chromatogram. Fig.17 The results show that the negative blank solution does not have a chromatographic peak corresponding to the test solution in the chromatogram, and there will be no interference.

[0131] Example 5: Preparation method of Jingangteng capsule test solution and optimization of chromatographic conditions

[0132] 1. Selection of extraction solvent

[0133] Take three portions of Jingangteng Capsule S10 (batch number: 20240105), about 2g each, and accurately add 50mL of methanol, 90% methanol, and 60% methanol, respectively, weigh the weight, and ultrasonically treat at a power of 500W and a frequency of 40kHz for 15min, cool, weigh again, make up the lost weight with the corresponding solvent, shake well, filter, and take the filtrate as the test solution, and determine it under the same method. Determine the best extraction solvent by comparison. The chromatograms are as follows Fig.18 (a) Fig.18 (b) Fig.18 As shown in (c), the chromatographic peaks of 60% methanol and methanol extraction are small and the extraction is insufficient, while the main chromatographic peak height of 90% methanol extraction is higher than that of methanol extraction, the extraction efficiency is high, and the peak shape is better. Therefore, 90% methanol was selected as the extraction solvent for preparing the Jingangteng Capsule test sample.

[0134] 2. Selection of extraction method

[0135] Take two portions of Jingangteng Capsule S10 (batch number: 20240105), about 2 g each, put them in a stoppered conical flask, accurately add 50 mL of 90% methanol, weigh the weight, and use ultrasonic treatment (power 200 W, frequency 40 kHz) for 15 min and reflux extraction for 15 min, respectively. Cool, weigh again, make up the lost weight with 90% methanol, shake well, filter, take the filtrate as the test solution, and determine it under the same method. Determine the best extraction method by comparison. The chromatograms are as follows: Fig.19 (a) Fig.19 (b) As shown in the figure, the main chromatographic peak height in the chromatogram of ultrasonic extraction for 15 min is higher than that of reflux extraction for 15 min. In order to ensure good extraction efficiency and prevent the degradation of chemical components during heating, ultrasonic treatment for 15 min was selected as the extraction method for preparing the Jingangteng capsule sample.

[0136] 3. Selection of extraction time

[0137] Take three portions of Jingangteng Capsule S10 (batch number: 20240105), about 2 g each, put them in a stoppered conical bottle, add 50 mL of 90% methanol, weigh them, and treat them with ultrasound (power 500 W, frequency 40 kHz) for 15 min, 30 min, and 60 min, cool them, weigh them again, make up the lost weight with 90% methanol, shake them well, filter them, and take the filtrate as the test solution. Determine the best extraction time by comparison under the same method, and the chromatograms are as follows: Fig. 20 (a) Fig. 20 (b) Fig. 20(c) As shown. Among them, there is no significant difference in the chromatograms of ultrasonic extraction for 15min and 30min, and the peak shape is good, while the area of ​​individual chromatographic peaks becomes smaller after ultrasonic treatment for 60min. Therefore, ultrasonic treatment for 15min was selected as the optimal extraction time for preparing Jingangteng Capsule test sample.

[0138] 4. Selection of mobile phase

[0139] Take an appropriate amount of Jingangteng Capsule S10 (batch number: 20240105), prepare it according to the test solution preparation method in Example 1, and compare the following mobile phases with a C18 chromatographic column: ① 0.1% phosphoric acid (A)-acetonitrile (B), ② 0.1% glacial acetic acid (A)-acetonitrile (B), ③ 0.1% formic acid (A)-acetonitrile (B), and perform gradient elution using the gradient elution program in Example 1. The chromatograms are as follows: Fig.21 (a) Fig.21 (b) Fig.21 As shown in (c), although the above three mobile phases all achieved separation, mobile phase ① had the best separation effect, while mobile phase ② had poor separation effect on peak 2 and the previous peak; and mobile phase ③ had lower response than mobile phase ①. Therefore, mobile phase ① was selected as the mobile phase for the Jingangteng Capsule test sample.

[0140] 5. Selection of detection wavelength

[0141] Take an appropriate amount of Jingangteng Capsule S10 (batch number: 20240105), prepare the test solution according to the preparation method of Example 1, inject it into the high performance liquid chromatograph for analysis, and follow the chromatographic conditions of Example 1, where the detection wavelengths are compared at 280nm, 303nm, and 326nm. The chromatograms are as follows: Fig. 22 (a) Fig. 22 (b) Fig. 22 (c) As shown. Among them, with 303nm as the detection wavelength, the chromatographic information is richer, the baseline is relatively stable, the response values ​​of each peak are relatively balanced, and it can relatively comprehensively reflect the component information in the sample, especially the peak 5 with lower response is larger at other wavelengths. Therefore, 303nm is selected as the detection wavelength of Jingangteng Capsule test sample.

[0142] 6. Selection of column temperature

[0143] Take an appropriate amount of Jingangteng Capsule S10 (batch number: 20240105), prepare the test solution according to the preparation method of Example 1, inject it into the high performance liquid chromatography for analysis, wherein the column temperature is 25°C, 30°C, and 35°C, respectively, and the separation of the chromatogram is as follows: Fig.23 (a) Fig.23 (b) Fig.23(c) When the column temperature is 25℃, the chromatographic peak separation effect is not good, especially peaks 2 and 4 are not separated from the nearby impurity peaks, which affects the accuracy of the results; when the column temperature is 30℃, the response values ​​of each peak are not as high as those at 35℃. Therefore, 35℃ is selected as the column temperature for the fingerprint of Jingangteng Capsule.

[0144] 7. Selection of chromatographic columns

[0145] Take an appropriate amount of Jingangteng Capsule S10 (batch number: 20240105), prepare the test solution according to the preparation method of Example 1, inject it into the high performance liquid chromatograph for analysis, and analyze it with chromatograms ① CAPCELL PAK C18, ② GL Science WondaSil C18, ③ NanoChrom ChromCoreC18, and ④ Waters SunFire C18, respectively. The chromatograms are as follows: Fig.24 (a) Fig.24 (b) Fig.24 (c) Fig.24 As shown in (d), it can be seen that when using ②GL Science WondaSil C18 column, peaks 2 and 6 are not separated from the preceding and following impurity peaks; using ①, ③, and ④ chromatographic columns can well separate the target components, indicating that this method has good adaptability to chromatographic columns of different brands.

[0146] Example 6: Determination of the contents of 8 ingredients in Jingangteng capsules

[0147] 1. Preparation of solution: same as in Example 1.

[0148] 2. Chromatographic conditions: same as in Example 1.

[0149] 3. Determination of ingredient content and methodology investigation

[0150] 3.1 Linear relationship investigation: Take appropriate amount of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol reference substances, weigh accurately, add 90% methanol to dissolve and prepare reference substance stock solutions containing neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol at 841 μg / mL, 2011 μg / mL, 1624 μg / mL, 521 μg / mL, 952 μg / mL, 530 μg / mL, 429 μg / mL and 515 μg / mL respectively. Dilute the above reference substance stock solutions step by step with 90% methanol to prepare series mixed standard solutions No. 1 to No. 8, and measure according to the chromatographic conditions under "2. Chromatographic conditions" and record the chromatogram. With the concentration of the linear solution series as the abscissa (x) and the corresponding peak area as the ordinate (y), a linear regression equation was drawn, and the linear equations and linear ranges of each component are shown in Table 9. As can be seen from Table 9, r is greater than 0.999, and each component has a good linear relationship within its respective range.

[0151] Table 9: Linear equations and linear ranges of 8 components

[0152]

[0153] 3.2 Precision test: Same as “3.1 Precision test” in above Example 4.

[0154] 3.3 Stability test: Same as “3.2 Stability test” in above Example 4.

[0155] 3.4 Repeatability test: Same as “3.3 Repeatability test” in above Example 4.

[0156] 3.5 Sample recovery test: 1g of the test sample of Jingangteng Capsule (S10) with known component content was accurately weighed, wherein the contents of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin, and resveratrol were 1.08mg / g, 2.58mg / g, 1.23mg / g, 0.17mg / g, 0.18mg / g, 0.68mg / g, 0.23mg / g, and 0.47mg / g, respectively. Accurately add each reference solution in a 1:1 ratio of each component in the sample, and repeat 6 times. Prepare the test solution according to the method under "1.2 Test Solution", and perform the sample injection and determination according to the chromatographic conditions under "2. Chromatographic Conditions". Calculate the average sample recovery of each component, and the results are shown in Table 10. The results showed that the average recoveries of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol were 103.15%, 94.55%, 96.79%, 92.08%, 99.87%, 92.77%, 96.44% and 92.23% (n=6), respectively. The recovery rate of this method was good.

[0157] Table 10: Recovery rates of 8 components in 15 batches of Jingangteng capsules

[0158]

[0159] 3.6 Sample content determination: Take the test solution of Jingangteng Capsules (S1~S15), and inject and determine it according to the chromatographic conditions under "2. Chromatographic conditions". Calculate the contents of the 8 components in the sample. The results are shown in Table 11.

[0160] Table 11: Content determination results of 8 components of 15 batches of Jingangteng capsules (mg / g)

[0161]

[0162] In summary, by optimizing the HPLC chromatographic conditions, the HPLC fingerprint of the Vajra vine preparation was successfully established, and a total of 8 main common peaks were identified. These 8 common peaks were attributed, and 8 active ingredients could be determined simultaneously; the similarity between the spectra of 15 batches of Vajra vine preparations and the control fingerprint was greater than 0.9, indicating that the types of chemical components contained in each batch of samples were basically the same, and there was good uniformity between the samples; this detection method is accurate, reliable, and simple to operate, and can provide a basis for the quality control of the preparation.

[0163] The above examples are merely illustrative of the present invention and do not constitute a limitation on the protection scope of the present invention. All designs that are the same or similar to the present invention fall within the protection scope of the present invention.

Claims

1. A method for establishing a fingerprint of a gypsophila preparation, characterized in that: The steps include: S1. Preparation of the test solution: weigh the Kanthus chinensis preparation, add methanol, perform ultrasonic treatment, and filter to obtain the test solution; S2. Preparation of reference solution: weigh 8 reference substances including cryptochlorogenic acid, neochlorogenic acid, chlorogenic acid, polydatin, astilbin, oxidized resveratrol, quercetin and resveratrol, mix them, add methanol to prepare a reference solution; S3. Establishment of fingerprint: The test solution and reference solution prepared as above were measured by HPLC respectively, the chromatograms were recorded and compared, the common peaks were identified, and the fingerprint of the scutellaria preparation was obtained.

2. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 1, characterized in that: In the step S1, the methanol concentration is 90%, the power of the ultrasonic treatment is 200 W, the frequency is 40 kHz, and the ultrasonic treatment time is 15 min.

3. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 1, characterized in that: The concentration of cryptochlorogenic acid in the reference solution is 2.190 μg / mL, the concentration of neochlorogenic acid is 18.914 μg / mL, the concentration of chlorogenic acid is 46.320 μg / mL, the concentration of polydatin is 1.661 μg / mL, the concentration of astilbin is 46.379 μg / mL, the concentration of oxidized resveratrol is 12.524 μg / mL, the concentration of quercetin is 46.411 μg / mL, and the concentration of resveratrol is 2.5967 μg / mL.

4. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 1, characterized in that: In step S3, the chromatographic conditions of HPLC are as follows: the chromatographic column is CAPCELL PAK C18, 250 mm×4.6 mm, 5 μm; the mobile phase is 0.1% phosphoric acid solution-acetonitrile, and gradient elution is performed; the detection wavelength is 200 nm to 400 nm; the flow rate is 1 mL / min; the column temperature is 25° C. to 40° C.; the injection volume is 10 μL; and the theoretical plate number calculated based on the chlorogenic acid peak should be no less than 10,000.

5. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 4, characterized in that: The chromatographic conditions of the HPLC are as follows: the chromatographic column is CAPCELL PAK C18, 250 mm×4.6 mm, 5 μm; the mobile phase is 0.1% phosphoric acid solution-acetonitrile, and gradient elution is performed; the detection wavelength is 303 nm; the flow rate is 1 mL / min; the column temperature is 35° C.; the injection volume is 10 μL; and the theoretical plate number calculated based on the chlorogenic acid peak should be no less than 10,000.

6. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 4 or 5, characterized in that: The gradient elution program is as follows by volume percentage: 0-5 min, 6% acetonitrile, 94% 0.1% phosphoric acid solution; 5-15 min, 6%-10% acetonitrile, 94%-90% 0.1% phosphoric acid solution; 15-60 min, 10%-29% acetonitrile, 90%-71% 0.1% phosphoric acid solution; 60-65 min, 29%-6% acetonitrile, 71%-94% 0.1% phosphoric acid solution; 65-70 min, 6% acetonitrile, 94% 0.1% phosphoric acid solution.

7. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 1, characterized in that: In step S3, the chromatogram of the test solution is imported into the "Chinese medicine chromatographic fingerprint similarity evaluation system software" to perform similarity evaluation with the chromatogram of the reference solution, determine the similarity, and determine the common peaks.

8. The method for establishing the fingerprint of the scutellaria baicalensis preparation according to claim 1, characterized in that: The gypsophila paniculata preparation is gypsophila paniculata syrup or gypsophila paniculata capsule.

9. A fingerprint obtained by the method for establishing the fingerprint of the Kanthus tricolor preparation according to any one of claims 1 to 8.

10. Application of the method for establishing the fingerprint of the Knotweed Herb preparation as described in any one of claims 1 to 8 in the determination of the content of ingredients in the Knotweed Herb preparation and the quality inspection of the Knotweed Herb preparation.

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