Method for establishing characteristic spectrum of Xiao'er Fengre Qing mixture and characteristic spectrum

Abstract

The present invention relates to a method for establishing a characteristic chromatogram of Xiao'er Fengre Qing mixture, comprising the following steps: (1) Preparation of a reference substance solution: Weigh appropriate amounts of chlorogenic acid reference substance, paeoniflorin reference substance, baicalin reference substance, phillyrin reference substance, arctiin reference substance, naringin reference substance, and neohesperidin reference substance, and prepare a solution; (2) Preparation of a test sample solution: Pipette Xiao'er Fengre Qing mixture, dilute, shake well, and filter to obtain the solution; (3) Determination: Use high performance liquid chromatography to separately detect the test sample solution and the reference substance solution, obtain the characteristic chromatogram of the test sample solution and the characteristic chromatogram of the reference substance solution, conduct qualitative analysis by comparing retention times, and conduct attribution and positioning of the target components, thereby obtaining the characteristic chromatogram of the pharmaceutical preparation.

Description

Technical Field

[0001] The present invention generally relates to the technical field of detection of components of traditional Chinese medicine, and in particular to a method for establishing a characteristic spectrum of a pediatric Fengreqing mixture and the characteristic spectrum. Background Art

[0002] The pediatric Fengreqing mixture is made from 20 herbs including honeysuckle, forsythia, isatis root, mint, bupleurum, burdock fruit, schizonepeta spicate, gypsum, scutellaria, gardenia, platycodon, red peony root, reed root, stir-fried bitter almond, bamboo leaf, fructus aurantii, stir-fried Liushenqu, bombyx batryticatus, saposhnikovia divaricata, and licorice. The herbs are decocted, concentrated, precipitated, filtered, and then bottled and sterilized using advanced modern pharmaceutical technology. It has the effects of relieving exterior symptoms with pungent and cool properties, clearing away heat and detoxifying, and relieving cough and sore throat. It is suitable for symptoms such as fever, cough, sputum, nasal congestion and runny nose, and red, swollen, and painful throat.

[0003] Due to the complex ingredients of Chinese patent medicines, a single indicator is difficult to fully control the quality of the product. Chinese patent medicines are a multi-component, multi-target system that emphasizes integrity. Therefore, establishing a characteristic spectrum that comprehensively reflects the main ingredients can more comprehensively reflect the intrinsic quality of Chinese patent medicines and comprehensively control and evaluate Chinese patent medicines from a holistic perspective.

[0004] The characteristic spectrum of traditional Chinese medicine is a chromatographic or spectral spectrum of a certain type of characteristic ingredients common to traditional Chinese medicines. It is holistic, fuzzy and quantifiable. As an effective quality control model, it has strong specificity and characteristic properties. It has become an effective method for quality control and evaluation of traditional Chinese medicines and has been recognized by the industry. Summary of the Invention

[0005] The purpose of the present invention is to provide a method for establishing a characteristic spectrum of pediatric Fengreqing mixture and a characteristic spectrum. The establishment method has the advantages of simple and rapid operation, low detection cost, good reproducibility and large amount of information, which provides a strong guarantee for the comprehensive and effective control of the quality of pediatric Fengreqing mixture; the characteristic spectrum provided has strong specificity and is easy to identify.

[0006] The technical solution of the present invention is to first provide a method for establishing a characteristic spectrum of Xiaoer Fengreqing Mixture, comprising the following steps:

[0007] S1. Preparation of reference solution: Take chemical substances corresponding to the components of Xiaoer Fengreqing mixture as reference substances, prepare them into solutions respectively, and obtain reference solution;

[0008] S2. Preparation of test solution: Take Xiaoer Fengreqing mixture, dilute with purified water, shake well, and filter to obtain the test solution;

[0009] S3, Chromatographic Conditions and System Suitability Test: Set the conditions of the high performance liquid chromatography system. The chromatographic column is packed with octadecylsilyl silica gel, and the mobile phase is one of acetonitrile - 0.1 vt% phosphoric acid aqueous solution, acetonitrile - water, and methanol - 0.1 vt% phosphoric acid. Gradient elution is used. Column temperature: 20 - 40 °C; UV detection wavelength 190 - 400 nm, time 90 min; injection volume 2 - 3 μl, preferably 2 μl.

[0010] S4, Determination: Use high performance liquid chromatography to detect the test solution and the reference solution respectively, obtain the characteristic chromatograms of the test solution and the reference solution, compare the retention times of the characteristic chromatograms of the test solution and the reference solution for qualitative analysis, and locate the target components in the characteristic chromatogram of the test solution, so as to obtain the characteristic chromatogram of the pharmaceutical preparation.

[0011] Further, in the above step S1, the reference substances include: chlorogenic acid reference substance, paeoniflorin reference substance, baicalin reference substance, forsythin reference substance, arctiin reference substance, naringin reference substance, neohesperidin reference substance. The method for preparing them into solutions is: accurately weigh each reference substance separately and dissolve them in methanol to make solutions containing 0.1 mg per 1 ml respectively.

[0012] Further, in the above step S2, the dilution means: Pipette 1.00 ml of Xiao'er Fengre Qing mixture, dilute it to 10 ml with purified water and shake well; the filtration uses a 0.22 μm aqueous microporous membrane for filtration.

[0013] Further, in the above step S3, the mobile phase is acetonitrile - 0.1 vt% phosphoric acid aqueous solution, where phase A is acetonitrile and phase B is 0.1 vt% phosphoric acid aqueous solution; column temperature: 30 °C; UV detection wavelength 230 nm.

[0014] In gradient elution, the chromatographic column is Thermo Accucore aQ, and a short column with a column length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 2.6 μm is used.

[0015] Further, in the above step S3, gradient elution is performed in the following manner: from 0 to 5 minutes, the volume percentage of mobile phase A is 3%, and the volume percentage of mobile phase B is 97%; from 5 to 10 minutes, the volume percentage of mobile phase A is increased from 3% to 5%, and the volume percentage of mobile phase B is decreased from 97% to 95%; from 10 to 20 minutes, the volume percentage of mobile phase A is 7%, and the volume percentage of mobile phase B is 93%; from 20 to 35 minutes, the volume percentage of mobile phase A is 15%, and the volume percentage of mobile phase B is 85%; from 35 to 55 minutes, the volume percentage of mobile phase A is 20%, and the volume percentage of mobile phase B is 80%; from 55 to 70 minutes, the volume percentage of mobile phase A is 50%, and the volume percentage of mobile phase B is 50%; from 70 to 80 minutes, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%; from 80 to 90 minutes, the volume percentage of mobile phase A is decreased from 100% to 3%, and the volume percentage of mobile phase B is increased from 0% to 97%.

[0016] Further, the above step S4 includes the following steps:

[0017] Step a: Prepare the test solution of Xiao'er Fengre Qingheji in different batches according to step S2, and perform high performance liquid chromatography detection to obtain the characteristic chromatograms of the test solutions of Xiao'er Fengre Qingheji in different batches. Then generate a common characteristic chromatogram, use the chromatographic peaks that exist in all the chromatograms as common characteristic peaks, determine the relative retention times of the common characteristic peaks, and establish the standard characteristic chromatogram of Xiao'er Fengre Qingheji;

[0018] Step b: Measure the characteristic chromatograms of each reference substance in step S1 to obtain the characteristic chromatogram of the reference substance solution;

[0019] Step c: Compare the established standard characteristic chromatogram of Xiao'er Fengre Qingheji with the characteristic chromatogram of the reference substance solution to locate each characteristic peak in the standard characteristic chromatogram of Xiao'er Fengre Qingheji. The specific positioning results are: the results of locating each characteristic peak in the standard characteristic chromatogram of Xiao'er Fengre Qingheji in step c are: it is determined that the 2nd peak is the characteristic peak of chlorogenic acid, the 6th peak is the characteristic peak of paeoniflorin, the 10th peak is the characteristic peak of naringin, the 13th peak is the characteristic peak of neohesperidin, the 16th peak is the characteristic peak of baicalin, the 17th peak is the characteristic peak of forsythin, and the 18th peak is the characteristic peak of arctiin.

[0020] Further, in the high performance liquid chromatography system of step S3, the detector is a diode array detector.

[0021] The present invention also provides a characteristic chromatogram of Xiao'er Fengre Qing mixture. It is obtained by the method for establishing the characteristic chromatogram of Xiao'er Fengre Qing mixture, and altogether includes 20 common peaks. The relative retention times of each common peak are respectively: 1 - 0.1822, 2 - 0.3425, 3 - 0.4150, 4 - 0.4381, 5 - 0.4670, 6 - 0.5556, 7 - 0.6397, 8 - 0.7359, 9 - 0.8086, 10 - 0.9270, 11 - 0.9503, 12 - 0.9618, 13 - 1.0000, 14 - 1.0363, 15 - 1.0475, 16 - 1.0994, 17 - 1.1410, 18 - 1.1565, 19 - 1.1867, 20 - 1.3430.

[0022] The present invention adopts the pre - treatment under optimized conditions and the high - performance liquid chromatography method to establish the characteristic chromatogram of Xiao'er Fengre Qing mixture, which can analyze 7 chemical components in Xiao'er Fengre Qing mixture, ensure the relative stability of the amount of effective components in the composition of Xiao'er Fengre Qing mixture, and thus ensure the safety and effectiveness of clinical medication. The method of the present invention can comprehensively and accurately evaluate the overall and internal quality of drug preparations.

[0023] The advantages of the present invention compared with the prior art are as follows:

[0024] Among the 20 medicinal materials in the prescription of Xiao'er Fengre Qing mixture, those that can be determined by a liquid - phase ultraviolet detector are: Flos Lonicerae, Fructus Forsythiae, Radix Isatidis, Radix Bupleuri, Fructus Arctii, Spica Schizonepetae, Radix Scutellariae, Fructus Gardeniae, Radix Paeoniae Rubra, and Semen Armeniacae Amarum Praeparatum. And those that can be detected by the liquid - chromatography elution procedure of the present invention are Flos Lonicerae, Radix Paeoniae Rubra, Radix Scutellariae, Fructus Forsythiae, Fructus Arctii, and Fructus Gardeniae. Several representative compounds are: chlorogenic acid, paeoniflorin, baicalin, phillyrin, arctiin, naringin, and neohesperidin. Through analysis, the spectral characteristics of each component are obvious. Identified and screened within the spectral range determined by the present invention, its chromatogram can reflect the information of most medicinal materials, and has a flat baseline, few interferences, and good separation. Description of the Drawings

[0025] From the following detailed description of the embodiments of the present invention in conjunction with the drawings, these and / or other aspects and advantages of the present invention will become clearer and easier to understand, wherein:

[0026] Figure 1 It shows the standard characteristic chromatogram of Xiao'er Fengre Qing mixture in the present invention;

[0027] Figure 2 It shows the characteristic chromatograms of 16 batches of different samples of Xiao'er Fengre Qing mixture. Detailed Embodiments

[0028] To enable those skilled in the art to better understand the present invention, the present invention will be further described in detail below in conjunction with the accompanying drawings and specific embodiments.

[0029] The reagents and instruments used in the embodiments of the present invention are as follows:

[0030] 1. The reagents used are shown in Table 1 below:

[0031] Table 1 Reagent names and characteristics used

[0032] Reagent Name Purity Manufacturer Batch Number Acetonitrile Chromatographic Grade Li'anlong Bohua (Tianjin) Pharmaceutical Chemical Co., Ltd. 20221011 Phosphoric Acid Analytical Reagent Grade Yantai Far East Fine Chemical Co., Ltd. 20210330

[0033] 2. The information of the reference substances is shown in Table 2 below:

[0034] Table 2 Reference substance names and characteristics

[0035] Name Manufacturer Batch Number Purity Naringin Reference Standard National Institutes for Food and Drug Control 110831-201906 91.5％ Neohesperidin Reference Standard National Institutes for Food and Drug Control 111857-201817 96.8％ Chlorogenic Acid Reference Standard National Institutes for Food and Drug Control 110753-201803 96.7％ Paeoniflorin Reference Standard National Institutes for Food and Drug Control 110736-201803 96.7％ Phillyrin Reference Standard National Institutes for Food and Drug Control 110821-201803 96.7％ Arctiin Reference Standard National Institutes for Food and Drug Control 110819-201812 95.0％ Baicalin Reference Standard National Institutes for Food and Drug Control 110715-202122 94.2％

[0036] 3. Instrument: High-performance liquid chromatograph, manufacturer: Waters, model: Acquity Arc, serial number: 6181B.

[0037] Example 1

[0038] A characteristic chromatogram of Xiao'er Fengre Qing Mixture and its establishment method, comprising the following steps:

[0039] 1. Preparation of the reference substance solution

[0040] Take appropriate amounts of chlorogenic acid reference substance, paeoniflorin reference substance, baicalin reference substance, forsythin reference substance, arctiin reference substance, naringin reference substance, and neohesperidin reference substance, accurately weigh them respectively, and prepare solutions containing 0.1 mg per 1 ml with methanol respectively;

[0041] 2. Preparation of the test solution

[0042] Precisely pipette 1.00 ml of Xiao'er Fengre Qing Mixture into a 10 ml volumetric flask, dilute to the mark with purified water, shake well, and filter through a 0.22 μm aqueous microporous membrane to obtain the solution.

[0043] 3. Chromatographic conditions and system suitability test: Set the conditions of the high-performance liquid chromatography system, wherein the chromatographic column is packed with octadecylsilane-bonded silica gel, and the mobile phase is one of acetonitrile - 0.1 vt% phosphoric acid aqueous solution, acetonitrile - water, and methanol - 0.1 vt% phosphoric acid, gradient elution is adopted, column temperature: 20 - 40 °C; ultraviolet detection wavelength 190 - 400 nm, time 90 min; injection volume 2 μl;

[0044] 4. Determination

[0045] The test solution and the reference solution were respectively detected by high performance liquid chromatography (HPLC) to obtain the characteristic chromatogram of the test solution and the characteristic chromatogram of the reference solution. The characteristic chromatogram of the test solution was compared with the characteristic chromatogram of the reference solution to qualitatively determine the retention time, and the index components in the characteristic chromatogram of the test solution were attributed and located, so as to obtain the characteristic chromatogram of the pharmaceutical preparation.

[0046] Among them, the high performance liquid chromatography includes the following detection conditions:

[0047] Octadecylsilane chemically bonded silica gel was used as the filler (column length: 100 mm, inner diameter: 2.1 mm). The methodological chromatographic column was Thermo Accucore aQ, with a column length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 2.6 μm; the column temperature was 30 °C, the detection wavelength was 230 nm, and the injection volume was 2 μl.

[0048] The mobile phase was tried with various ratios of acetonitrile - 0.1 vt% phosphoric acid, acetonitrile - water, and methanol - 0.1 vt% phosphoric acid at different gradients to explore the separation conditions. Finally, it was determined that acetonitrile was used as mobile phase A and 0.1% phosphoric acid solution was used as mobile phase B, and the gradient elution was carried out according to the regulations in Table 3 below, with the best effect.

[0049] Table 3 Liquid phase elution program: Acetonitrile as mobile phase A and 0.1 vt% phosphoric acid solution as mobile phase B

[0050]

[0051] According to the above method, 12 batches of Xiao'er Fengre Qingheji were detected to obtain the characteristic chromatograms of multiple Xiao'er Fengre Qingheji samples and generate a common characteristic chromatogram. The chromatographic peaks that all existed in the chromatograms were used as common characteristic peaks, and the relative retention times of the common characteristic peaks were determined to establish the standard characteristic chromatogram of Xiao'er Fengre Qingheji.

[0052] As Figure 1 shown, the characteristic chromatogram of Xiao'er Fengre Qingheji includes 20 characteristic peaks. Taking the 13th peak as the reference peak (S peak, retention time: 1.0000), the relative retention times of the other 19 peaks are respectively: 1 - 0.1822, 2 - 0.3425, 3 - 0.4150, 4 - 0.4381, 5 - 0.4670, 6 - 0.5556, 7 - 0.6397, 8 - 0.7359, 9 - 0.8086, 10 - 0.9270, 11 - 0.9503, 12 - 0.9618, 14 - 1.0363, 15 - 1.0475, 16 - 1.0994, 17 - 1.1410, 18 - 1.1565, 19 - 1.1867, 20 - 1.3430.

[0053] As Figure 1As shown, the standard characteristic chromatogram of Xiao'er Fengre Qing mixture was compared with the characteristic chromatogram of the reference solution. Seven peaks could be identified in the reference solution, and the identification results are shown in Table 4.

[0054] Table 4 Identification of characteristic peaks in the reference solution

[0055]

[0056] The detection method of the characteristic chromatogram of Xiao'er Fengre Qing mixture of the present invention was subjected to methodological verification, and the results of its performance indicators are as follows.

[0057] 1. Precision

[0058] Precisely pipette 1 ml of the 03222015 batch sample of Xiao'er Fengre Qing mixture, and dilute it to 10 ml with water to prepare the test solution. Take 2 μl of the above solution and inject it into the high-performance liquid chromatograph, and continuously measure 9 times for the precision test. The results are shown in Table 5. The results show that the RSD of the relative retention time ≤ 0.1575%, proving that the precision of this method is good.

[0059] Table 5 Precision test results of the relative retention time of 20 characteristic peaks in the characteristic chromatogram

[0060]

[0061] 2. Repeatability

[0062] Precisely pipette 1 ml each of 9 portions of the 03222015 batch sample of Xiao'er Fengre Qing mixture, and dilute each portion to 10 ml with water to prepare the test solutions. Respectively take 2 μl of the above test solutions and inject them into the high-performance liquid chromatograph for determination for the repeatability test. The results are shown in Table 6. The results show that the RSD of the relative retention time ≤ 0.1591%, indicating that the repeatability of this method is good.

[0063] Table 6 Repeatability test results of the relative retention time of 20 characteristic peaks in the characteristic chromatogram

[0064]

[0065] 3. Stability

[0066] Precisely pipette 1 ml of the 03222015 batch sample of Xiao'er Fengre Qing mixture, and dilute it to 10 ml with water to prepare the test solution. Let it stand at room temperature, and measure it at regular intervals. Calculate the relative retention time of 20 chromatographic characteristic peaks, and measure until 35 hours. The results are shown in Table 7 below. It can be seen that the contents of 14 components to be measured in the test solution are stable for at least 35 hours.

[0067] Table 7 Stability test results of the characteristic chromatogram of Xiao'er Fengre Qing mixture

[0068]

[0069] 4. Determination of multi-batch samples

[0070] Prepare 16 batches of Xiao'er Fengre Qing Heji samples according to the method in the main text. Prepare the test solution according to the preparation method of the test sample. Respectively draw 2 μl of the above test solution and inject it into the high performance liquid chromatograph for determination. The results are as Figure 2 , shown in Table 8. The results show that the RSD of the relative retention time ≤ 0.1738%, the selected characteristic peaks are stably reproduced in multi-batch samples, and there is no difference among different batches of Xiao'er Fengre Qing Heji, indicating that the process is stable.

[0071] Table 8 for Xiao'er Fengreqing Mixture Test results of differences among batches of characteristic chromatograms

[0072]

[0073] The embodiments of the present invention have been described above. The above description is exemplary and not exhaustive, and is not limited to the disclosed embodiments. Many modifications and changes are obvious to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. A method for establishing a characteristic fingerprint of Xiao'er Fengre Qing Mixture, characterized in that, The following steps are involved: S1. Preparation of reference solution: chemical substances corresponding to the components of Xiaoer Fengreqing mixture are taken as reference substances, and are prepared into solutions respectively to obtain reference solution; the reference substances include: chlorogenic acid reference substance, paeoniflorin reference substance, baicalin reference substance, forsythin reference substance, arctiin reference substance, naringin reference substance, and neohesperidin reference substance; S2. Preparation of test solution: Take Xiaoer Fengreqing mixture, dilute with purified water, shake well, and filter to obtain the test solution; S3. Chromatographic conditions and system suitability experiment: HPLC system conditions were set up, with the chromatographic column packed with octadecylsilane bonded silica gel, the mobile phase consisting of acetonitrile-0.1vt% phosphoric acid aqueous solution, and gradient elution. The chromatographic column was ThermoAccucore aQ, with a short column length of 100 mm, an inner diameter of 2.1 mm, and a particle size of 2.6 μm. The column temperature was 20-40°C, the UV detection wavelength was 230 nm, and the run time was 90 min. The injection volume was 2-3 μl, and the gradient elution degree was: From 0 to 5 min, the volume percentage of mobile phase A is 3%, and the volume percentage of mobile phase B is 97%; From 5 to 10 min, the volume percentage of mobile phase A increased from 3% to 5%, and the volume percentage of mobile phase B decreased from 97% to 95%; From 10 to 20 min, the volume percentage of mobile phase A increased from 5% to 7%, and the volume percentage of mobile phase B decreased from 95% to 93%; From 20 to 35 min, the volume percentage of mobile phase A increased from 7% to 15%, and the volume percentage of mobile phase B decreased from 93% to 85%; From 35 to 55 min, the volume percentage of mobile phase A increased from 15% to 20%, and the volume percentage of mobile phase B decreased from 85% to 80%; From 55 to 70 min, the volume percentage of mobile phase A increased from 20% to 50%, and the volume percentage of mobile phase B decreased from 80% to 50%; From 70 to 75 min, the volume percentage of mobile phase A increased from 50% to 100%, and the volume percentage of mobile phase B decreased from 50% to 0%; From 75 to 80 min, the volume percentage of mobile phase A is 100%, and the volume percentage of mobile phase B is 0%; From 80 to 82 min, the volume percentage of mobile phase A decreased from 100% to 3%, and the volume percentage of mobile phase B increased from 0% to 97%; From 82 to 90 min, the volume percentage of mobile phase A was 3%, and the volume percentage of mobile phase B was 97%; S4. Determination: Use high performance liquid chromatography to detect the test solution and the reference solution respectively, obtain the characteristic spectrum of the test solution and the characteristic spectrum of the reference solution, compare the retention time of the characteristic spectrum of the test solution with that of the reference solution for qualitative analysis, attribute and locate the indicator components in the characteristic spectrum of the test solution, and thus obtain the characteristic spectrum of the pharmaceutical preparation.

2. The method for establishing the characteristic fingerprint of Xiao'er Fengre Qing mixture as described in claim 1, characterized in that Step S1, the preparation method of the reference substance solution is: accurately weigh each reference substance respectively, and add methanol to make a solution containing 0.1 mg of reference substance per 1 ml.

3. The method for establishing the characteristic fingerprint of Xiao'er Fengre Qing mixture as described in claim 1, characterized in that In step S2, the dilution means: pipette 1.00 ml of Xiao'er Fengre Qing mixture, dilute it to 10 ml with purified water and shake well; the filtration uses a 0.22 μm water-based microporous membrane for filtration.

4. The method for establishing the characteristic fingerprint of Xiao'er Fengre Qing mixture as described in claim 1, characterized in that In step S3, the mobile phase is acetonitrile - 0.1 vt% phosphoric acid aqueous solution, wherein, phase A is acetonitrile and phase B is 0.1 vt% phosphoric acid aqueous solution; column temperature: 30 °C; ultraviolet detection wavelength 230 nm; the injection volume in high performance liquid chromatography is 2 μL.

5. The method for establishing the characteristic fingerprint of Xiao'er Fengre Qing mixture as described in claim 1, characterized in that Step S4 includes the following steps: Step a, prepare the test solution of Xiao'er Fengre Qing mixture of different batches according to step S2, and perform high performance liquid chromatography detection to obtain the characteristic fingerprints of the test solutions of Xiao'er Fengre Qing mixture of different batches, then generate a common characteristic fingerprint, use the chromatographic peaks that all exist in the fingerprint as the common characteristic peaks, determine the relative retention time of the common characteristic peaks, and establish the standard characteristic fingerprint of Xiao'er Fengre Qing mixture; Step b, measure the characteristic fingerprints of each reference substance in step S1 to obtain the characteristic fingerprint of the reference substance solution; Step c, compare the established standard characteristic fingerprint of Xiao'er Fengre Qing mixture with the characteristic fingerprint of the reference substance solution to locate each characteristic peak in the standard characteristic fingerprint of Xiao'er Fengre Qing mixture.

6. The method for establishing the characteristic chromatogram of the Xiao'er Fengre Qing mixture as described in claim 5, characterized in that, The result of locating each characteristic peak in the standard characteristic fingerprint of Xiao'er Fengre Qing mixture in step c is: determine that peak 2 is the characteristic peak of chlorogenic acid, peak 6 is the characteristic peak of paeoniflorin, peak 10 is the characteristic peak of naringin, peak 13 is the characteristic peak of neohesperidin, peak 16 is the characteristic peak of baicalin, peak 17 is the characteristic peak of forsythin, and peak 18 is the characteristic peak of arctiin.

7. The method for establishing the characteristic fingerprint of Xiao'er Fengre Qing mixture as described in claim 1, characterized in that In the high performance liquid chromatography system of step S3, the detector is a diode array detector.

8. The method for establishing the characteristic chromatogram of the Xiao'er Fengre Qing mixture according to claim 1, characterized in that, The fingerprint includes a total of 20 common peaks, and the relative retention times of each common peak are respectively: 1 - 0.1822, 2 - 0.3425, 3 - 0.4150, 4 - 0.4381, 5 - 0.4670, 6 - 0.5556, 7 - 0.6397, 8 - 0.7359, 9 - 0.8086, 10 - 0.9270, 11 - 0.9503, 12 - 0.9618, 13 - 1.0000, 14 - 1.0363, 15 - 1.0475, 16 - 1.0994, 17 - 1.1410, 18 - 1.1565, 19 - 1.1867, 20 - 1.3430.

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