Fingerprint spectrum of Xiaoer daochi tablet, construction method thereof, determination method of index component content and application thereof

Abstract

The present application relates to a kind of pediatric guide red slice fingerprint construction method, the construction method includes using high performance liquid chromatography to determine the index component in pediatric guide red slice;Wherein, the extraction solvent of the pediatric guide red slice is methanol-water, the chromatographic column of the high performance liquid chromatography uses reversed-phase C 18 Chromatographic column, the index component includes gallic acid, gardenoside, catechin, aloe emodin, rhein, emodin, chrysophanol and emodin methyl ether.The technical scheme of the present application optimizes the extraction mode of the effective component of pediatric guide red slice, can effectively extract the effective component in preparation, and establishes the construction method of pediatric guide red slice fingerprint by high performance liquid chromatography, the liquid phase fingerprint of pediatric guide red slice can be obtained by the method, and the index component of pediatric guide red slice can be effectively characterized.

Description

Technical Field

[0001] This invention relates to the field of fingerprinting of traditional Chinese medicine, and in particular to a fingerprint of Xiaodaochi tablets and its construction method, as well as the determination method and application of its index component content. Background Technology

[0002] Xiaodaochi Tablets are a commonly used traditional Chinese medicine preparation in pediatrics. Its functions and indications are clearing heat and promoting bowel movements. It is mainly used for symptoms such as gastrointestinal heat accumulation, oral ulcers, sore throat, bleeding gums, swollen and painful cheeks, acute conjunctivitis, constipation, and dark yellow urine. The specific prescription composition and manufacturing process are as follows: 1. Prescription: Rhubarb 270g, Gardenia 216g, Akebia 72g, Talc 72g, Rehmannia 44g, Licorice 72g, Poria 72g; 2. Manufacturing process: The above seven ingredients are prepared by pulverizing rhubarb and talc into fine powder, sifting, and setting aside. Gardenia, Akebia, Rehmannia, Licorice, and Poria are decocted twice with water, and the decoctions are combined and concentrated into a paste. The concentrated paste is then mixed with the rhubarb powder and other excipients, granulated, dried, and compressed into tablets.

[0003] Currently, research on Xiaodaochi tablets is limited. Existing technologies for content determination mainly focus on single-component analysis of rhein, emodin, and geniposide, lacking fingerprint analysis and multi-component evaluation studies involving multiple herbs. Furthermore, the current quality standard for Xiaodaochi tablets is the national ministerial standard for traditional Chinese medicine preparations, Volume 19, WS3-B-3537-98, which primarily covers properties, thin-layer chromatography identification of rhubarb, and routine tablet inspections, lacking content determination. This makes it difficult to comprehensively and accurately reflect the overall quality of the preparation. In addition, the sources of Chinese medicinal herbs are complex and diverse, and the lack of unified and detailed standards for planting, harvesting, and processing easily leads to inconsistent quality of medicinal materials, thus affecting the final quality and efficacy of Xiaodaochi tablets.

[0004] Therefore, there is an urgent need in this field to construct a method for building fingerprint spectra of pediatric daphne tablets that is simple to operate, has good stability and reproducibility, and can comprehensively and accurately evaluate the quality of pediatric daphne tablets. Summary of the Invention

[0005] To address the aforementioned problems in the existing technology, the purpose of this invention is to provide a fingerprint spectrum of Xiaodaochi tablets and its construction method, as well as a method and application for determining the content of its indicator components. This is used for the detection and quality control of the effective components of Xiaodaochi tablets. The fingerprint spectrum of Xiaodaochi tablets constructed by this method can provide a more comprehensive quality evaluation of Xiaodaochi tablets, improve the efficiency of quality detection, and is simple, accurate, and reproducible.

[0006] In a first aspect, the present invention provides a method for constructing a fingerprint spectrum of Xiaodaochi tablets, the method comprising determining indicator components in Xiaodaochi tablets using high-performance liquid chromatography (HPLC); wherein the extraction solvent of Xiaodaochi tablets is methanol-water, and the chromatographic column of the HPLC method is a reversed-phase C24 column. 18 The chromatographic column contains the index components including gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

[0007] Preferably, the indicator components further include glycyrrhizin and / or ammonium glycyrrhizate.

[0008] Preferably, the volume ratio of methanol to water is 10:90 to 90:10, more preferably 60:40 to 90:10, and even more preferably 75:25.

[0009] Preferably, the construction method includes the following steps: (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms.

[0010] Preferably, the extraction is ultrasonic extraction; more preferably, the extraction time is 15-45 minutes.

[0011] Preferably, the antiphase C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm.

[0012] Preferably, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution; more preferably, the mobile phase A of the high performance liquid chromatography is acetonitrile, and the mobile phase B is 0.1% (v / v) phosphoric acid aqueous solution.

[0013] Preferably, the high-performance liquid chromatography method includes gradient elution using the mobile phase; more preferably, the gradient elution procedure is as follows: .

[0014] Preferably, the flow rate of the mobile phase is 0.5~1.5 mL / min, more preferably 0.9~1.1 mL / min, and even more preferably 1.0 mL / min.

[0015] Preferably, the column temperature of the high-performance liquid chromatography is 25~45℃, and more preferably 35℃.

[0016] Preferably, the injection volume of the high-performance liquid chromatography is 5~20μL, and more preferably 10μL.

[0017] Preferably, the detection wavelength of the high-performance liquid chromatography is 210~255nm, more preferably one of 215nm, 223nm, 230nm, 238nm, and 251nm, and more preferably 230nm.

[0018] Preferably, the pediatric fingerprint spectrum contains 27 common peaks, of which peak 26 is a reference peak, and the retention times of each common peak include: Peak 2, with an average retention time (RT) of 6.143–6.190 min; Peak 4, with an average retention time (RT) of 13.240–13.350 min; Peak 5, with an average retention time (RT) of 14.093–14.177 min; Peak 22, with an average retention time (RT) of 58.487–58.660 min; Peak 24, with an average retention time (RT) of 64.493–64.590 min; Peak 25, with an average retention time (RT) of 69.886–69.987 min; Peak 26, with an average retention time (RT) of 70.570–70.680 min; Peak 27, with an average retention time (RT) of 73.687–73.860 min.

[0019] Preferably, the retention time of each common peak further includes: Peak 8, with an average retention time (RT) of 24.463–24.590 min; Peak 23, with an average retention time (RT) of 62.800–62.950 min.

[0020] Preferably, the retention time of each common peak further includes: Peak 1, with an average retention time (RT) of 4.583–4.617 min; Peak 3, with an average retention time (RT) of 11.103–11.200 min; Peak 6, with an average retention time (RT) of 22.677–22.797 min; Peak 7, with an average retention time (RT) of 23.283–23.413 min; Peak 9, with an average retention time (RT) of 25.187–25.352 min; Peak 10 had an average retention time (RT) of 26.773–26.937 min. Peak 11, with an average retention time (RT) of 33.360–33.527 min; Peak 12, with an average retention time (RT) of 34.470–34.640 min; Peak 13, with an average retention time (RT) of 36.037–36.213 min; Peak 14, with an average retention time (RT) of 39.547–39.707 min; Peak 15, with an average retention time (RT) of 40.880–41.037 min; Peak 16 had an average retention time (RT) of 42.393–42.533 min. Peak 17, with an average retention time (RT) of 43.937–44.305 min; Peak 18, with an average retention time (RT) of 47.420–47.547 min; Peak 19, with an average retention time (RT) of 49.250–49.415 min; Peak 20 had an average retention time (RT) of 50.567–50.687 min. Peak 21 had an average retention time (RT) of 55.377–55.467 min.

[0021] Secondly, the present invention provides a pediatric fingerprint spectrum constructed by any of the above-described construction methods.

[0022] Thirdly, the present invention provides a method for determining the content of indicator components in Xiaodaochi tablets, the method comprising determining the indicator components in Xiaodaochi tablets using high performance liquid chromatography; wherein the extraction solvent of Xiaodaochi tablets is methanol-water, and the chromatographic column of the high performance liquid chromatography is a reversed-phase C24 column. 18 The chromatographic column contains two or more of the following index components: gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

[0023] Preferably, the indicator components further include glycyrrhizin and / or ammonium glycyrrhizate.

[0024] Preferably, the volume ratio of methanol to water is 10:90 to 90:10, more preferably 60:40 to 90:10, and even more preferably 75:25.

[0025] Preferably, the determination method includes the following steps.

[0026] (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms.

[0027] Preferably, the extraction is ultrasonic extraction; more preferably, the extraction time is 15-45 minutes.

[0028] Preferably, the antiphase C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm.

[0029] Preferably, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution; more preferably, the mobile phase A of the high performance liquid chromatography is acetonitrile, and the mobile phase B is 0.1% (v / v) phosphoric acid aqueous solution.

[0030] Preferably, the high-performance liquid chromatography method includes gradient elution using the mobile phase; more preferably, the gradient elution procedure is as follows: .

[0031] Preferably, the flow rate of the mobile phase is 0.5~1.5 mL / min, more preferably 0.9~1.1 mL / min, and even more preferably 1.0 mL / min.

[0032] Preferably, the column temperature of the high-performance liquid chromatography is 25~45℃, and more preferably 35℃.

[0033] Preferably, the injection volume of the high-performance liquid chromatography is 5~20μL, and more preferably 10μL.

[0034] Preferably, the detection wavelength of the high-performance liquid chromatography is 210~255nm, and more preferably one of 215nm, 223nm, 238nm, and 251nm.

[0035] Preferably, the detection wavelength of gallic acid is 215 nm.

[0036] Preferably, the detection wavelength of geniposide is 238 nm.

[0037] Preferably, the detection wavelength of the catechin is 238 nm.

[0038] Preferably, the detection wavelength of the aloe-emodin is 223 nm.

[0039] Preferably, the detection wavelength of rhein is 223 nm.

[0040] Preferably, the detection wavelength of the emodin is 223 nm.

[0041] Preferably, the detection wavelength of the rhein is 223 nm.

[0042] Preferably, the detection wavelength of the emodin methyl ether is 223 nm.

[0043] Preferably, the detection wavelength of the glycyrrhizin is 215 nm.

[0044] Preferably, the detection wavelength of the ammonium glycyrrhizate is 251 nm.

[0045] Fourthly, the present invention provides an application of the construction method described in any one of the above claims, the fingerprint spectrum of the pediatric daphne tablet described in any one of the above claims, or the determination method described in any one of the above claims in the quality detection of pediatric daphne tablets.

[0046] Preferably, the quality testing includes one or more of the following: identification of indicator components, determination of the content of indicator components, and batch quality consistency evaluation.

[0047] The beneficial effects of this invention are as follows: 1. The method for constructing the fingerprint chromatogram of Xiaodaochi tablets of the present invention, based on the structural characteristics and physicochemical properties of the active ingredients contained in Xiaodaochi tablets, screened and optimized the analytical conditions such as the sample processing method, mobile phase, detection wavelength, elution program, column temperature, and flow rate, and completed the systematic methodological validation. The obtained fingerprint chromatogram of Xiaodaochi tablets has high resolution, good peak shape, and good baseline separation for each characteristic chromatographic peak. It has good stability and many characteristic peaks, and can comprehensively, accurately, and intuitively evaluate the quality of Xiaodaochi tablets, making it suitable for the quality control of Xiaodaochi tablets.

[0048] 2. This invention uses HPLC wavelength switching method to simultaneously identify 10 components in the characteristic spectrum and determines the content of 10 components, including gallic acid, geniposide, catechin, glycyrrhizin, aloe-emodin, glycyrrhizic acid ammonium, rhein, emodin, chrysophanol, and emodin methyl ether, thereby achieving comprehensive evaluation and control of the quality of Xiaordaochi tablets.

[0049] 3. The fingerprint spectrum of Xiaodaochi tablets constructed in this invention overcomes the shortcomings of current standards, such as single control indicators, low specificity, and low quality standard levels. The constructed fingerprint spectrum contains 27 common peaks, identifying and attributing 10 specific components, enabling a comprehensive evaluation of the quality of Xiaodaochi tablets and effectively ensuring the quality of the finished product. Simultaneously, it can serve as an internal control standard for large-scale production enterprises and an evaluation basis for the consistency of Xiaodaochi tablet efficacy, providing assurance for the safe, effective, uniform, and stable standardized production of Xiaodaochi tablets.

[0050] 4. The method for constructing the fingerprint spectrum of Xiaodaochi tablets and the method for determining the content of indicator components in Xiaodaochi tablets of this invention are simple, fast, accurate, and reproducible, requiring no extraction, concentration, or evaporation. They are easy to master and widely adopted, effectively improving detection efficiency, reducing detection costs, and minimizing environmental pollution. Under the same elution conditions, by switching wavelengths, it is possible to simultaneously determine 10 indicator components and construct fingerprint spectra, with good peak separation, achieving the goals of multi-evaluation with one measurement, high efficiency, low loss, and easy popularization.

[0051] 5. The method of this invention not only provides a method for the simultaneous detection of multiple medicinal materials (rhubarb, gardenia, and licorice) in the prescription of Xiaodaochi tablets, but also provides a reference and research ideas for the simultaneous determination of these components in other compound preparations.

[0052] 6. The method of this invention optimizes the extraction of the effective components of Xiaodaochi tablets, effectively extracting the effective components from the preparation. Furthermore, a high-performance liquid chromatography (HPLC) method was established to construct the fingerprint spectrum of Xiaodaochi tablets. This method can obtain the liquid phase fingerprint spectrum of Xiaodaochi tablets, effectively characterizing the indicator components. The fingerprint spectrum of this invention can rapidly and accurately identify the authenticity and homogeneity of the finished product, which is of great significance for comprehensively controlling product quality. It also has advantages such as simple operation, high specificity, good repeatability, good stability, and high precision. Attached Figure Description

[0053] Figure 1 Example 3: 1. Chromatogram obtained in blank solvent.

[0054] Figure 2 Example 3: 2. Chromatogram obtained by extending the elution time.

[0055] Figure 3 Example 3: 3. In the identification and labeling of common peaks in fingerprint chromatograms, the chromatograms obtained by matching and superimposing fingerprint chromatograms of 22 batches of Xiaodaochi tablets were used.

[0056] Figure 4 For Example 3:4. In the confirmation of the reference peak, the chromatogram obtained from the mixed reference standard.

[0057] Figure 5 For Example 3: 4. In the confirmation of the reference peak, the chromatogram of the S1 pediatric erythrocyte lacryma-jobi sample.

[0058] Figure 6 Example 3:5. In the construction of the fingerprint spectrum of Xiaodaochi tablets, the fingerprint spectrum of Xiaodaochi tablets is used.

[0059] Figure 7 For Example 3: 6. Chromatograms obtained from mixed reference standards in specificity test.

[0060] Figure 8 For Example 3: 6. chromatogram obtained from the S1 pediatric chamomile tablet sample in the specificity test.

[0061] Figure 9 For Example 3: 6. Chromatogram obtained from rhubarb negative sample in specificity test.

[0062] Figure 10 Example 3: 6. Chromatogram obtained from a gardenia negative sample in the specificity test.

[0063] Figure 11 For Example 3: 6. Chromatogram obtained from licorice negative sample in specificity test.

[0064] Figure 12 For Example 3: 6. Chromatogram obtained from Rehmannia glutinosa negative sample in specificity test.

[0065] Figure 13 Example 3: 6. Chromatogram obtained from a negative sample of Akebia quinata in the specificity test.

[0066] Figure 14 Example 3: 6. Chromatogram obtained from a negative sample of Poria cocos in the specificity test.

[0067] Figure 15 Example 4: 1. Ultraviolet spectra of gallic acid, geniposide, catechin, glycyrrhizin, aloe-emodin, ammonium glycyrrhizate, rhein, emodin, chrysophanol, and emodin methyl ether in the linear range test.

[0068] Figure 16 The chromatogram obtained from the mixed reference standard (215nm) in the determination of the content of the index components in Xiaodaochi tablets in Example 4: 6.

[0069] Figure 17 The chromatogram obtained from the mixed reference standard (238nm) in the determination of the content of the index components in Xiaodaochi tablets in Example 4: 6.

[0070] Figure 18The chromatogram obtained from the mixed reference standard (223 nm) in the determination of the content of the index components in Xiaodaochi tablets in Example 4: 6.

[0071] Figure 19 The chromatogram obtained from the mixed reference standard (251 nm) in the determination of the content of the index components in Xiaodaochi tablets in Example 4: 6.

[0072] Figure 20 For Example 4:6. In the determination of the content of index components in Xiaodaochi tablets, the chromatogram obtained from sample S1 (215nm) of Xiaodaochi tablets.

[0073] Figure 21 For Example 4:6. In the determination of the content of index components in Xiaodaochi tablets, the chromatogram obtained from sample S1 (238nm) of Xiaodaochi tablets.

[0074] Figure 22 For Example 4:6. In the determination of the content of index components in Xiaodaochi tablets, the chromatogram obtained from sample S1 (223nm) of Xiaodaochi tablets.

[0075] Figure 23 For Example 4:6. In the determination of the content of index components in Xiaodaochi tablets, the chromatogram obtained from sample S1 (251nm) of Xiaodaochi tablets. Detailed Implementation

[0076] To make the technical solution and beneficial effects of the present invention more apparent and understandable, a detailed description is provided below by listing specific embodiments. The accompanying drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application pertains.

[0077] As used herein, the term “fingerprint” refers to a characteristic spectrum or image of a group of components obtained by spectral or chromatographic determination.

[0078] As used in this article, the term "mobile phase system" refers to the substance that carries the analyte forward during chromatography.

[0079] In a first aspect, the present invention provides a method for constructing a fingerprint spectrum of Xiaodaochi tablets, the method comprising determining indicator components in Xiaodaochi tablets using high-performance liquid chromatography (HPLC); wherein the extraction solvent of Xiaodaochi tablets is methanol-water, and the chromatographic column of the HPLC method is a reversed-phase C24 column. 18 The chromatographic column contains the index components including gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

[0080] In some embodiments, the indicator components further include glycyrrhizin and / or ammonium glycyrrhizate.

[0081] In some embodiments, the indicator components include gallic acid, geniposide, catechin, glycyrrhizin, aloe-emodin, ammonium glycyrrhizate, rhein, emodin, chrysophanol, and emodin methyl ether.

[0082] In some embodiments, the volume ratio of methanol to water is 10:90 to 90:10.

[0083] In some embodiments, the volume ratio of methanol to water is 60:40 to 90:10.

[0084] In some embodiments, the volume ratio of methanol to water is 75:25.

[0085] In some implementations, the construction method includes the following steps: (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms.

[0086] In some embodiments, the extraction is ultrasonic extraction.

[0087] In some implementations, the extraction time is 15 to 45 minutes.

[0088] In some implementations, the extraction time is 30 minutes.

[0089] In some embodiments, the inverted C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm.

[0090] In some embodiments, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution.

[0091] In some embodiments, the mobile phase system of the high-performance liquid chromatography is acetonitrile-0.1% (v / v) phosphoric acid aqueous solution.

[0092] In some embodiments, the mobile phase A of the high-performance liquid chromatography is acetonitrile, and the mobile phase B is a 0.1% (v / v) aqueous solution of phosphoric acid.

[0093] In some embodiments, the high-performance liquid chromatography method includes gradient elution using the mobile phase; In some implementations, the gradient elution procedure is as follows: .

[0094] In some embodiments, the flow rate of the mobile phase is 0.5 to 1.5 mL / min.

[0095] In some embodiments, the flow rate of the mobile phase is 0.9 to 1.1 mL / min.

[0096] In some embodiments, the flow rate of the mobile phase is 1.0 mL / min.

[0097] In some embodiments, the column temperature of the high-performance liquid chromatography is 25~45°C.

[0098] In some embodiments, the column temperature of the high-performance liquid chromatography is 35°C.

[0099] In some embodiments, the injection volume of the high-performance liquid chromatography is 5~20 μL.

[0100] In some embodiments, the injection volume of the high-performance liquid chromatography is 10 μL.

[0101] In some embodiments, the detection wavelength of the high-performance liquid chromatography is 210~255nm.

[0102] In some embodiments, the detection wavelength of the high-performance liquid chromatography is one of 215 nm, 223 nm, 230 nm, 238 nm, and 251 nm.

[0103] In some embodiments, the detection wavelength of the high-performance liquid chromatography is 230 nm.

[0104] In some embodiments, the pediatric fingerprint spectrum contains 27 common peaks, of which peak 26 is a reference peak, and the retention times of each common peak include: Peak 2, with an average retention time (RT) of 6.143–6.190 min; Peak 4, with an average retention time (RT) of 13.240–13.350 min; Peak 5, with an average retention time (RT) of 14.093–14.177 min; Peak 22, with an average retention time (RT) of 58.487–58.660 min; Peak 24, with an average retention time (RT) of 64.493–64.590 min; Peak 25, with an average retention time (RT) of 69.886–69.987 min; Peak 26, with an average retention time (RT) of 70.570–70.680 min; Peak 27, with an average retention time (RT) of 73.687–73.860 min.

[0105] In some embodiments, the fingerprint spectrum of the pediatric daphne tablet contains 27 common peaks, of which peak 26 is the reference peak, peak 2 is gallic acid, peak 4 is geniposide, peak 5 is catechin, peak 22 is aloe-emodin, peak 24 is rhein, peak 25 is emodin, peak 26 is chrysophanol, and peak 27 is emodin methyl ether.

[0106] In some implementations, the retention time of each common peak further includes: Peak 8, with an average retention time (RT) of 24.463–24.590 min; Peak 23, with an average retention time (RT) of 62.800–62.950 min.

[0107] In some implementations, the retention time of each common peak further includes: Peak 1, with an average retention time (RT) of 4.583–4.617 min; Peak 3, with an average retention time (RT) of 11.103–11.200 min; Peak 6, with an average retention time (RT) of 22.677–22.797 min; Peak 7, with an average retention time (RT) of 23.283–23.413 min; Peak 9, with an average retention time (RT) of 25.187–25.352 min; Peak 10 had an average retention time (RT) of 26.773–26.937 min. Peak 11, with an average retention time (RT) of 33.360–33.527 min; Peak 12, with an average retention time (RT) of 34.470–34.640 min; Peak 13, with an average retention time (RT) of 36.037–36.213 min; Peak 14, with an average retention time (RT) of 39.547–39.707 min; Peak 15, with an average retention time (RT) of 40.880–41.037 min; Peak 16 had an average retention time (RT) of 42.393–42.533 min. Peak 17, with an average retention time (RT) of 43.937–44.305 min; Peak 18, with an average retention time (RT) of 47.420–47.547 min; Peak 19, with an average retention time (RT) of 49.250–49.415 min; Peak 20 had an average retention time (RT) of 50.567–50.687 min. Peak 21 had an average retention time (RT) of 55.377–55.467 min.

[0108] In some embodiments, the pediatric fingerprint spectrum contains 27 common peaks, of which peak 26 is a reference peak, and the retention times of each common peak include: Peak 2, with an average retention time (RT) of 6.143–6.190 min; Peak 4, with an average retention time (RT) of 13.240–13.350 min; Peak 5, with an average retention time (RT) of 14.093–14.177 min; Peak 8, with an average retention time (RT) of 24.463–24.590 min; Peak 22, with an average retention time (RT) of 58.487–58.660 min; Peak 23, with an average retention time (RT) of 62.800–62.950 min.

[0109] Peak 24, with an average retention time (RT) of 64.493–64.590 min; Peak 25, with an average retention time (RT) of 69.886–69.987 min; Peak 26, with an average retention time (RT) of 70.570–70.680 min; Peak 27, with an average retention time (RT) of 73.687–73.860 min.

[0110] In some embodiments, the fingerprint spectrum of the pediatric dermatitis tablets contains 27 common peaks, of which peak 26 is the reference peak, peak 2 is gallic acid, peak 4 is geniposide, peak 5 is catechin, peak 8 is glycyrrhizin, peak 22 is aloe-emodin, peak 23 is ammonium glycyrrhizate, peak 24 is rhein, peak 25 is emodin, peak 26 is chrysophanol, and peak 27 is emodin methyl ether.

[0111] In some embodiments, the pediatric fingerprint spectrum contains 27 common peaks, of which peak 26 is a reference peak, and the retention times of each common peak include: Peak 1, with an average retention time (RT) of 4.583–4.617 min; Peak 2, with an average retention time (RT) of 6.143–6.190 min; Peak 3, with an average retention time (RT) of 11.103–11.200 min; Peak 4, with an average retention time (RT) of 13.240–13.350 min; Peak 5, with an average retention time (RT) of 14.093–14.177 min; Peak 6, with an average retention time (RT) of 22.677–22.797 min; Peak 7, with an average retention time (RT) of 23.283–23.413 min; Peak 8, with an average retention time (RT) of 24.463–24.590 min; Peak 9, with an average retention time (RT) of 25.187–25.352 min; Peak 10 had an average retention time (RT) of 26.773–26.937 min. Peak 11, with an average retention time (RT) of 33.360–33.527 min; Peak 12, with an average retention time (RT) of 34.470–34.640 min; Peak 13, with an average retention time (RT) of 36.037–36.213 min; Peak 14, with an average retention time (RT) of 39.547–39.707 min; Peak 15, with an average retention time (RT) of 40.880–41.037 min; Peak 16 had an average retention time (RT) of 42.393–42.533 min. Peak 17, with an average retention time (RT) of 43.937–44.305 min; Peak 18, with an average retention time (RT) of 47.420–47.547 min; Peak 19, with an average retention time (RT) of 49.250–49.415 min; Peak 20 had an average retention time (RT) of 50.567–50.687 min. Peak 21, with an average retention time (RT) of 55.377–55.467 min; Peak 22, with an average retention time (RT) of 58.487–58.660 min; Peak 23, with an average retention time (RT) of 62.800–62.950 min; Peak 24, with an average retention time (RT) of 64.493–64.590 min; Peak 25, with an average retention time (RT) of 69.886–69.987 min; Peak 26, with an average retention time (RT) of 70.570–70.680 min; Peak 27, with an average retention time (RT) of 73.687–73.860 min.

[0112] Secondly, the present invention provides a pediatric fingerprint spectrum constructed by any of the above-described construction methods.

[0113] Thirdly, the present invention provides a method for determining the content of indicator components in Xiaodaochi tablets, the method comprising determining the indicator components in Xiaodaochi tablets using high performance liquid chromatography; wherein the extraction solvent of Xiaodaochi tablets is methanol-water, and the chromatographic column of the high performance liquid chromatography is a reversed-phase C24 column. 18 The chromatographic column contains two or more of the following index components: gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

[0114] In some embodiments, the indicator components further include glycyrrhizin and / or ammonium glycyrrhizate.

[0115] In some embodiments, the indicator components also include glycyrrhizin and ammonium glycyrrhizate.

[0116] In some embodiments, the volume ratio of methanol to water is 10:90 to 90:10.

[0117] In some embodiments, the volume ratio of methanol to water is 60:40 to 90:10.

[0118] In some embodiments, the volume ratio of methanol to water is 75:25.

[0119] In some embodiments, the determination method includes the following steps.

[0120] (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms.

[0121] In some embodiments, the extraction is ultrasonic extraction.

[0122] In some implementations, the extraction time is 15 to 45 minutes.

[0123] In some implementations, the extraction time is 30 minutes.

[0124] In some embodiments, the inverted C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm.

[0125] In some embodiments, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution.

[0126] In some embodiments, the mobile phase system of the high-performance liquid chromatography is acetonitrile-0.1% (v / v) phosphoric acid aqueous solution.

[0127] In some embodiments, the mobile phase A of the high-performance liquid chromatography is acetonitrile, and the mobile phase B is a 0.1% (v / v) aqueous solution of phosphoric acid.

[0128] In some embodiments, the high-performance liquid chromatography method includes gradient elution using the mobile phase.

[0129] In some implementations, the gradient elution procedure is as follows: .

[0130] In some embodiments, the flow rate of the mobile phase is 0.5 to 1.5 mL / min.

[0131] In some embodiments, the flow rate of the mobile phase is 0.9 to 1.1 mL / min.

[0132] In some embodiments, the flow rate of the mobile phase is 1.0 mL / min.

[0133] In some embodiments, the column temperature of the high-performance liquid chromatography is 25~45°C.

[0134] In some embodiments, the column temperature of the high-performance liquid chromatography is 35°C.

[0135] In some embodiments, the injection volume of the high-performance liquid chromatography is 5~20 μL.

[0136] In some embodiments, the injection volume of the high-performance liquid chromatography is 10 μL.

[0137] In some embodiments, the detection wavelength of the high-performance liquid chromatography is 210~255nm.

[0138] In some embodiments, the detection wavelength of the high-performance liquid chromatography is one of 215 nm, 223 nm, 238 nm, and 251 nm.

[0139] In some embodiments, the detection wavelength of gallic acid is 215 nm.

[0140] In some embodiments, the detection wavelength of geniposide is 238 nm.

[0141] In some embodiments, the detection wavelength of the catechin is 238 nm.

[0142] In some embodiments, the detection wavelength of the aloe-emodin is 223 nm.

[0143] In some embodiments, the detection wavelength of the rhein is 223 nm.

[0144] In some embodiments, the detection wavelength of the emodin is 223 nm.

[0145] In some embodiments, the detection wavelength of the rhein is 223 nm.

[0146] In some embodiments, the detection wavelength of the emodin methyl ether is 223 nm.

[0147] In some embodiments, the detection wavelength of the glycyrrhizin is 215 nm.

[0148] In some embodiments, the detection wavelength of the ammonium glycyrrhizate is 251 nm.

[0149] Fourthly, the present invention provides an application of the construction method described in any one of the above claims, the fingerprint spectrum of the pediatric daphne tablet described in any one of the above claims, or the determination method described in any one of the above claims in the quality detection of pediatric daphne tablets.

[0150] In some embodiments, the quality testing includes one or more of the following: identification of indicator components, determination of the content of indicator components, and batch quality consistency evaluation.

[0151] In some implementations, the quality testing includes the identification of the active ingredient.

[0152] Unless otherwise stated, experimental methods in the following examples without specific conditions are generally performed under standard conditions or as recommended by the manufacturer. Unless otherwise stated, all percentages, ratios, proportions, or parts are by weight. Unless otherwise stated, the room temperature described in this invention is 20°C to 30°C. Unless otherwise stated, the various materials and reagents used in this invention can be obtained using conventional methods in the art or through commercial channels, including but not limited to the following instruments and reagents: U3000 high performance liquid chromatograph (Thermo Fisher Scientific, Inc., USA); XSE 205DU electronic analytical balance (Mettler-Toledo International GmbH, Switzerland); WUC-D22H Ultrasonic Cleaner (Daihan Science Co., Ltd.); Methanol (chromatographic grade, Merck, Germany); Acetonitrile (chromatographic grade, Merck, Germany); 0.45µm organic phase filter membrane (Tianjin Jinteng Experimental Equipment Co., Ltd.)

[0153] Wahaha purified drinking water (Hangzhou Wahaha Group Co., Ltd.); all other reagents were of analytical grade.

[0154] The reference standards used in the embodiments of the present invention are those shown in Table 1.

[0155] Table 1. Reference Standard Information Table

[0156] The samples used in this embodiment of the invention are 22 batches of Xiaodaochi tablets produced by 3 manufacturers, as detailed in Table 2, Sample Information Table of Xiaodaochi Tablets.

[0157] Table 2. Sample Information Table of Pediatric Inhibitory Tablets

[0158] Note: " / " indicates that the specifications are not specified in the drug's instructions.

[0159] Example 1: Preparation of the test solution Take 20 tablets of this product sample, accurately weigh them, grind them into a fine powder, take a single dose (4 tablets), accurately weigh them, place them in a stoppered conical flask, accurately add 25 mL of 75% methanol solution, weigh them, sonicate for 30 min (power: 250 W; frequency: 40 kHz), cool to room temperature, weigh them again, replenish the lost weight with 75% methanol solution, shake well, filter, take the filtrate, filter it through a 0.45 μm microporous membrane to obtain the corresponding test solution (corresponding to the sample solution of Xiaodaochi tablets).

[0160] Example 2: Preparation of the reference solution Accurately weigh 10.11 mg of emodin reference standard, 10.43 mg of emodin methyl ether reference standard, 10.89 mg of rhein reference standard, 13.61 mg of aloe-emodin reference standard, 26.59 mg of glycyrrhizin reference standard, 24.70 mg of glycyrrhizic acid reference standard, 22.50 mg of gallic acid reference standard, and 24.42 mg of chrysophanol reference standard. Place them in a 250 mL volumetric flask, dissolve and dilute to the mark with methanol, and shake well. This is solution A.

[0161] In addition, accurately weigh 10.05 mg of catechin reference standard and 12.45 mg of geniposide reference standard, place them in a 25 mL volumetric flask, dissolve and dilute to the mark with the above solution A, shake well, and the mixed reference standard solution is obtained, as detailed in Table 3.

[0162] Table 3. Formulation of Mixed Reference Solution

[0163] Example 3: Construction of fingerprint spectrum of pediatric daphne tablets In this embodiment, high-performance liquid chromatography (HPLC) was used for determination. The chromatographic conditions were as follows: the chromatographic column was a Warters XBridgeShield RP. 18 (4.6×250mm, 5μm); the mobile phase was acetonitrile (A)-0.1% phosphoric acid solution (B), the flow rate was 1.0mL / min; the column temperature was 35℃; the detection wavelength was 230nm; the injection volume was 10μL; the gradient elution program is shown in Table 4.

[0164] Table 4. Gradient elution procedure

[0165] 1. Blank solvent To eliminate interference from the blank solvent, the above elution procedure was used, with a 75% methanol solution as the solvent injected, and the chromatogram was recorded. The results showed no solvent interference; see details below. Figure 1 .

[0166] 2. Extend the wash-off time In this embodiment, the gradient elution program time was 85 min. To verify whether a chromatographic peak would appear after 85 min, the above elution program was used to extend the rinsing time by 40 min. The chromatogram showed no chromatographic peak after 120 min. See details. Figure 2 .

[0167] 3. Identification and labeling of common peaks in fingerprint spectra Samples of Xiaodaochi Pian (a traditional Chinese medicine) S1-S22 were processed according to the method in Example 1, and the corresponding test solutions (corresponding Xiaodaochi Pian sample solutions) were extracted. Samples were also processed according to the method in Example 2, and the corresponding mixed reference solutions were extracted. The test solutions and mixed reference solutions were injected into a high-performance liquid chromatograph (HPLC) for analysis. The recorded chromatographic data were imported into the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (version 2012.130723). Using the chromatogram of sample solution S1 of Xiaodaochi Pian as a reference chromatogram, the mean method was used, with a time window width of 0.1 min. Multi-point correction and Mark peak matching were performed to generate a fingerprint chromatogram matching overlay and a fingerprint chromatogram (R). A total of 27 common peaks were identified. See details... Figure 3 .

[0168] 4. Confirmation of the reference peak By comparing the chromatograms of the mixed reference standard, the chromatogram of the S1 Xiaodaochi tablet sample, and the fingerprint chromatograms, the retention times and UV spectra of each component peak were identified, resulting in 10 common known peaks: gallic acid (peak 2), geniposide (peak 4), catechin (peak 5), glycyrrhizin (peak 8), aloe-emodin (peak 22), ammonium glycyrrhizate (peak 23), rhein (peak 24), emodin (peak 25), chrysophanol (peak 26), and emodin methyl ether (peak 27). Chrysophanol (peak 26), the indicator component of rhubarb (the principal ingredient), which has a relatively high content, was selected as the reference peak (S). See details... Figures 4 to 6 The relative retention times of each common peak are shown in Table 5.

[0169] Table 5. Relative retention times of each common peak

[0170] 5. Similarity evaluation of fingerprint spectra of pediatric diaphragm tablets The fingerprint spectrum was generated using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (version 2012.130723) through Mark peak matching. See details below. Figure 6 According to the similarity evaluation system for chromatographic fingerprints of traditional Chinese medicine, the similarity between the fingerprint chromatograms of the test samples and the fingerprint chromatograms after 4 minutes was calculated to be no less than 0.92. The similarity results of each batch of test samples and fingerprint chromatograms are detailed in Table 6.

[0171] Table 6. Similarity between fingerprint chromatograms of each batch of test samples

[0172] 6. Specificity test Take all the raw materials and excipients from the prescription, and prepare negative samples for products lacking rhubarb, gardenia, akebia stem, rehmannia root, licorice root, and poria cocos according to the process and sample solution preparation method of this product. Extract one portion of each negative sample according to the extraction method in Example 1. Accurately pipette 10 μL each of the mixed reference solution from Example 2, the S1 pediatric erythrocyte sedimentation tablet sample solution, and the negative sample solution, and inject them into the liquid chromatograph. Determine the chromatographic conditions of high performance liquid chromatography according to Example 3, as detailed in [link to Example 3]. Figures 7-14 The attribution of 27 common peaks was studied, and the results are detailed in Table 7.

[0173] Table 7. Results of the study on the attribution of common peaks

[0174] Note: " / " indicates that the ingredient was not detected.

[0175] The results showed that there were no interfering peaks at the retention times of gallic acid, geniposide, catechin, glycyrrhizin, aloe-emodin, ammonium glycyrrhizate, rhein, emodin, chrysophanol, and emodin methyl ether.

[0176] Example 4: Determination of the content of indicator components in Xiaodaochi tablets and methodological validation In this embodiment, high performance liquid chromatography (HPLC) was used for determination. The chromatographic conditions were as follows: column: Warters X Bridge Shield RP18 (4.6 × 250 mm, 5 µm); mobile phase: acetonitrile (A) - 0.1% phosphoric acid solution (B); flow rate: 1.0 mL / min; column temperature: 35 °C; detection wavelengths: 215 nm (gallic acid, glycyrrhizin), 238 nm (geniposide, catechin), 251 nm (ammonium glycyrrhizate), 223 nm (aloe-emodin, rhein, emodin, chrysophanol, emodin methyl ether); injection volume: 10 μL; gradient elution program is shown in Table 4 of Example 3.

[0177] 1. Linear range test Accurately pipette 0.1, 0.5, 1.2, 2.5, 5.0, 7.5, and 10.0 mL of the mixed reference solution from Example 2 into 10 mL volumetric flasks, and dilute to the mark with 75% methanol solution. Perform high-performance liquid chromatography (HPLC) determination, plotting peak area (Y) as the ordinate and sample concentration (X, μg·mL⁻¹) as the modulus. ‒1 Linear regression analysis was performed with the x-axis as the abscissa to obtain the regression equations for each analyte. The linear correlation data are shown in Table 8, and the UV absorption spectra are shown in [Table 8]. Figure 15 ).

[0178] Table 8. Linear Correlation

[0179] 2. Precision test Accurately pipette 10 μL of the mixed reference solution from step 1 (linear range test), inject it in six consecutive parallel injections, and perform high-performance liquid chromatography (HPLC) to determine the peak area. Record the peak area and calculate the RSD based on the peak area. The results show that the method has good precision, and the results are shown in Table 9.

[0180] Table 9. Precision (peak area) test results

[0181] 3. Repeatability test Sample S1 was processed according to the method in Example 1, and six test solutions were extracted. 10 µL of each test solution was precisely pipetted into the liquid chromatograph and analyzed by high-performance liquid chromatography (HPLC). The peak area was recorded and the content (unit: mg / g) was calculated. The results showed that the method had good repeatability, and the calculation results are shown in Table 10.

[0182] Table 10. Repeatability test results (unit: mg / g)

[0183] 4. Stability test Sample S1 was processed according to the method in Example 1. One sample solution was extracted and injected at 0h, 2h, 4h, 8h, 12h, and 24h for high-performance liquid chromatography (HPLC). The peak areas were recorded, and the RSD was calculated based on the peak areas. The results showed that the sample solution was stable within 24 hours, and the method had good stability. The results are shown in Table 11.

[0184] Table 11. Stability (peak area) test results

[0185] 5. Recovery rate test Six portions of sample S1, each approximately 0.36 g, were accurately weighed and 3.0 mL of the mixed reference solution from Example 2 was added to each portion. The mixture was then processed according to the method in Example 1. The six test solutions were extracted and analyzed by high performance liquid chromatography. The peak areas were recorded and the contents and recoveries were calculated. The results of the recovery tests for each component are shown in Table 12.

[0186] Table 12. Results of recovery tests for each component

[0187] 6. Determination of the content of indicator components in Xiaodaochi tablets Preparation of the test solution: It was prepared according to the method in Example 1.

[0188] Preparation of the reference solution: It was prepared according to the method in Example 2.

[0189] Determination by high performance liquid chromatography: Accurately pipette 10 μL each of the reference solution and the test solution, inject them into the liquid chromatograph, and perform high performance liquid chromatography to determine the result.

[0190] For detailed measurement results, please refer to [link / reference]. Figures 16-23 And Table 13.

[0191] Table 13. Results of content determination of Xiaodaochi tablets (mg / g)

[0192] It should be understood that the above embodiments are exemplary and are not intended to encompass all possible implementations included in the claims. Various modifications and changes can be made to the above embodiments without departing from the scope of this disclosure. Similarly, the various technical features of the above embodiments can be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only illustrate several implementations of the present invention and do not limit the scope of protection of this patent.

Claims

1. A method for constructing a fingerprint spectrum of pediatric diaphragm, characterized in that, The construction method includes determining the indicator components in Xiaodaochi tablets using high-performance liquid chromatography (HPLC); wherein the extraction solvent for Xiaodaochi tablets is methanol-water, and the chromatographic column used in the HPLC is a reversed-phase C24 column. 18 The chromatographic column contains the index components including gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

2. The construction method according to claim 1, characterized in that, The indicator components also include glycyrrhizin and / or ammonium glycyrrhizate; Preferably, the volume ratio of methanol to water is 10:90 to 90:10, more preferably 60:40 to 90:10, and even more preferably 75:

25.

3. The construction method according to claim 1 or 2, characterized in that, The construction method includes the following steps: (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms; Preferably, the extraction is ultrasonic extraction; more preferably, the extraction time is 15-45 minutes. Preferably, the antiphase C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm; Preferably, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution; more preferably, the mobile phase A of the high performance liquid chromatography is acetonitrile, and the mobile phase B is 0.1% (v / v) phosphoric acid aqueous solution. Preferably, the high-performance liquid chromatography method includes gradient elution using the mobile phase; more preferably, the gradient elution procedure is as follows: ； Preferably, the flow rate of the mobile phase is 0.5~1.5 mL / min, more preferably 0.9~1.1 mL / min, and even more preferably 1.0 mL / min; Preferably, the column temperature of the high-performance liquid chromatography is 25~45℃, and more preferably 35℃; Preferably, the injection volume of the high-performance liquid chromatography is 5~20 μL, more preferably 10 μL; Preferably, the detection wavelength of the high-performance liquid chromatography is 210~255nm, more preferably one of 215nm, 223nm, 230nm, 238nm, and 251nm, and more preferably 230nm.

4. The construction method according to any one of claims 1 to 3, characterized in that, The fingerprint spectrum of the pediatric diaphragm contains 27 common peaks, of which peak 26 is the reference peak. The retention times of each common peak are as follows: Peak 2, with an average retention time (RT) of 6.143–6.190 min; Peak 4, with an average retention time (RT) of 13.240–13.350 min; Peak 5, with an average retention time (RT) of 14.093–14.177 min; Peak 22, with an average retention time (RT) of 58.487–58.660 min; Peak 24, with an average retention time (RT) of 64.493–64.590 min; Peak 25, with an average retention time (RT) of 69.886–69.987 min; Peak 26, with an average retention time (RT) of 70.570–70.680 min; Peak 27, with an average retention time (RT) of 73.687–73.860 min; Preferably, the retention time of each common peak further includes: Peak 8, with an average retention time (RT) of 24.463–24.590 min; Peak 23, with an average retention time (RT) of 62.800–62.950 min; Preferably, the retention time of each common peak further includes: Peak 1, with an average retention time (RT) of 4.583–4.617 min; Peak 3, with an average retention time (RT) of 11.103–11.200 min; Peak 6, with an average retention time (RT) of 22.677–22.797 min; Peak 7, with an average retention time (RT) of 23.283–23.413 min; Peak 9, with an average retention time (RT) of 25.187–25.352 min; Peak 10 had an average retention time (RT) of 26.773–26.937 min. Peak 11, with an average retention time (RT) of 33.360–33.527 min; Peak 12, with an average retention time (RT) of 34.470–34.640 min; Peak 13, with an average retention time (RT) of 36.037–36.213 min; Peak 14, with an average retention time (RT) of 39.547–39.707 min; Peak 15, with an average retention time (RT) of 40.880–41.037 min; Peak 16 had an average retention time (RT) of 42.393–42.533 min. Peak 17, with an average retention time (RT) of 43.937–44.305 min; Peak 18, with an average retention time (RT) of 47.420–47.547 min; Peak 19, with an average retention time (RT) of 49.250–49.415 min; Peak 20 had an average retention time (RT) of 50.567–50.687 min. Peak 21 had an average retention time (RT) of 55.377–55.467 min.

5. A pediatric fingerprint spectrum constructed by the construction method of any one of claims 1 to 4.

6. A method for determining the content of indicator components in pediatric diaphoretic tablets, characterized in that, The determination method includes using high-performance liquid chromatography (HPLC) to determine the indicator components in Xiaodaochi tablets; wherein the extraction solvent for Xiaodaochi tablets is methanol-water, and the chromatographic column used in the HPLC is a reversed-phase C24 column. 18 The chromatographic column contains two or more of the following index components: gallic acid, geniposide, catechin, aloe-emodin, rhein, emodin, chrysophanol, and emodin methyl ether.

7. The determination method according to claim 6, characterized in that, The indicator components also include glycyrrhizin and / or ammonium glycyrrhizate; Preferably, the volume ratio of methanol to water is 10:90 to 90:10, more preferably 60:40 to 90:10, and even more preferably 75:

25.

8. The determination method according to claim 6 or 7, characterized in that, The determination method includes the following steps: (1) Preparation of test solution: After extracting the pediatric red tablets with the extraction solvent, dilute and filter with the extraction solvent to obtain the test solution; (2) Preparation of reference solution: Dissolve and dilute the reference standard of the indicator component using the extraction solvent to obtain the reference solution; (3) Determination by high performance liquid chromatography: Accurately measure the test solution and the reference solution, inject them into the liquid chromatograph respectively, and record the chromatograms; Preferably, the extraction is ultrasonic extraction; more preferably, the extraction time is 15-45 minutes. Preferably, the antiphase C 18 The chromatographic column was a Warters XBridge Shield RP18, 4.6 mm × 250 mm, 5 µm; Preferably, the mobile phase system of the high performance liquid chromatography is acetonitrile-(0.025~0.2%) (v / v) phosphoric acid aqueous solution; more preferably, the mobile phase A of the high performance liquid chromatography is acetonitrile, and the mobile phase B is 0.1% (v / v) phosphoric acid aqueous solution. Preferably, the high-performance liquid chromatography method includes gradient elution using the mobile phase; more preferably, the gradient elution procedure is as follows: ； Preferably, the flow rate of the mobile phase is 0.5~1.5 mL / min, more preferably 0.9~1.1 mL / min, and even more preferably 1.0 mL / min; Preferably, the column temperature of the high-performance liquid chromatography is 25~45℃, and more preferably 35℃; Preferably, the injection volume of the high-performance liquid chromatography is 5~20 μL, more preferably 10 μL; Preferably, the detection wavelength of the high-performance liquid chromatography is 210~255nm, and more preferably one of 215nm, 223nm, 238nm, and 251nm.

9. The determination method according to claim 8, characterized in that, The detection wavelength for gallic acid is 215 nm; Preferably, the detection wavelength of geniposide is 238 nm; Preferably, the detection wavelength of the catechin is 238 nm; Preferably, the detection wavelength of the aloe-emodin is 223 nm; Preferably, the detection wavelength of rhein is 223 nm; Preferably, the detection wavelength of the emodin is 223 nm; Preferably, the detection wavelength of the rhein is 223 nm; Preferably, the detection wavelength of the emodin methyl ether is 223 nm; Preferably, the detection wavelength of glycyrrhizin is 215 nm; Preferably, the detection wavelength of the ammonium glycyrrhizate is 251 nm.

10. The application of the construction method of any one of claims 1 to 4, the fingerprint spectrum of the pediatric dermatitis tablets as described in claim 5, or the determination method of any one of claims 6 to 9 in the quality detection of pediatric dermatitis tablets; Preferably, the quality testing includes one or more of the following: identification of indicator components, determination of the content of indicator components, and batch quality consistency evaluation.

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