Pharmaceutical composition as well as construction method and detection method of characteristic chromatogram of pharmaceutical composition

Abstract

The invention provides a pharmaceutical composition and a construction method of a characteristic spectrum of the pharmaceutical composition. The characteristic peak in the characteristic spectrum constructed by the method is used as a measurement index, the pharmaceutical composition can be accurately identified, the component content of the pharmaceutical composition can be determined, the specificity is strong, and the quality control of the pharmaceutical composition is facilitated. Moreover, the construction method is simple and convenient to operate, has the advantages of good reproducibility, high precision, strong stability and the like, and is suitable for popularization and application.

Description

Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a pharmaceutical composition in the field of traditional Chinese medicine and a construction method and a detection method of its characteristic spectrum. Background Art

[0002] For some products in the field of traditional Chinese medicine, the improvement and improvement of their preparation process may have an impact on product quality, including properties, physical and chemical identification, thin layer identification, routine inspection, content determination, etc. The quality standards of different batches of products need to be unified and need to be further improved. Therefore, establishing a comprehensive and quantifiable comprehensive ingredient quality monitoring method is important for improving product quality stability and consistency. Summary of the invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art to a certain extent. To this end, the present invention proposes a method for constructing a characteristic spectrum of a pharmaceutical composition, a method for identifying the pharmaceutical composition, a method for detecting the pharmaceutical composition, and a method for quality control of the pharmaceutical composition. The characteristic peaks in the characteristic spectrum of the pharmaceutical composition constructed by the method of the present application are used as a measurement index, which can accurately identify the pharmaceutical composition and determine the component content of the pharmaceutical composition, has strong specificity, and is helpful for the quality control of the pharmaceutical composition. In addition, the construction method is simple to operate, has the advantages of good reproducibility, high precision and strong stability, and is suitable for popularization and application.

[0004] In one aspect of the present invention, a method for constructing a characteristic spectrum of a pharmaceutical composition is proposed. It is characterized in that it includes: providing a plurality of different batches of pharmaceutical composition samples; pre-treating each sample under the same conditions to obtain a plurality of test solutions; performing high performance liquid chromatography on the plurality of test solutions under the same conditions to obtain a plurality of chromatograms; comparing and analyzing the plurality of chromatograms to determine the common characteristic peaks, so as to obtain the characteristic spectrum of the pharmaceutical composition; wherein the mobile phase used in the high performance liquid chromatography detection includes: mobile phase A, the mobile phase A is a 0.05-0.15 mass % phosphoric acid solution; mobile phase B, the mobile phase B is acetonitrile; the elution mode of the mobile phase is gradient elution; wherein the pharmaceutical composition The composition comprises an active ingredient, and the active ingredient is made from the following Chinese medicinal raw materials: honeysuckle, forsythia, Scrophularia, Isatis root, red peony root, scutellaria, mulberry leaf, chrysanthemum, peucedanum, bitter almond, burdock fruit, oriental rhizome, sterculia lychnophora, fried silkworm, cicada shell, and oroxylum butterfly; preferably, the weight ratio of any one raw material selected from honeysuckle, forsythia, Scrophularia, Isatis root to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, peucedanum, bitter almond, burdock fruit, oriental rhizome, sterculia lychnophora, silkworm, cicada shell, and oroxylum butterfly is 2.5:1, and the weight ratio of scutellaria baicalensis to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, peucedanum, bitter almond, burdock fruit, oriental rhizome, sterculia lychnophora, silkworm, cicada shell, and oroxylum butterfly is 1:1. Specifically, in the pharmaceutical composition, the active ingredient is made of the following Chinese medicinal raw materials in the following weight percentages: based on the total weight of the Chinese medicinal raw materials, 11.112% honeysuckle, 11.112% forsythia, 11.112% scrophularia, 11.112% isatis root, 4.444% red peony root, 6.668% scutellaria, 4.444% mulberry leaf, 4.444% chrysanthemum, 4.444% peucedanum, 4.444% bitter almond, 4.444% arctium, 4.444% oriental water plantain, 4.444% sterculia lychnophora, 4.444% bombyx batryticatus, 4.444% cicada shell, and 4.444% oroxylum butterfly.

[0005] The inventors of the present invention used high performance liquid chromatography to detect the pharmaceutical composition and found that the composition and elution conditions of the mobile phase would significantly affect the separation effect. When gradient elution was performed using 0.05-0.15 mass % phosphoric acid solution as mobile phase A and acetonitrile as mobile phase B, more chromatographic peaks with high separation and good peak shape could be obtained.

[0006] Furthermore, the inventors performed the same pretreatment and high-performance liquid chromatography detection on multiple different batches of samples, and determined 11 common characteristic peaks by comparing and analyzing the multiple chromatograms obtained. These 11 common characteristic peaks constitute the characteristic spectrum of the pharmaceutical composition, which can be used to identify the pharmaceutical composition, determine the component content of the pharmaceutical composition and whether the quality meets the standard, and achieve quality control. In addition, the method is simple to operate, has the advantages of good reproducibility, strong specificity, high precision and strong stability, and is suitable for promotion and application.

[0007] According to a specific embodiment of the present invention, the elution mode of the mobile phase is as follows:

[0008] Time (minutes) Mobile phase A (%) Mobile phase B (%) 0～5 90 10 5～15 90～80 10～20 15～50 80～65 20～35 50～55 65～40 35～60 55～65 40 60

[0009] The inventors obtained the above-mentioned preferred elution mode and flow rate through a large number of experiments, thereby all common characteristic peaks can be detected simultaneously, and a characteristic spectrum with high separation and good peak shape can be obtained.

[0010] According to a specific embodiment of the present invention, the flow rate of the mobile phase used in the high performance liquid chromatography detection is 0.8-1.2 mL / min, the column temperature is 25-35°C, and the wavelength is 200-250 nm. The inventors obtained the above-mentioned preferred chromatographic conditions through a large number of experiments, thereby all common characteristic peaks can be detected at the same time, and a characteristic spectrum with high separation and good peak shape can be obtained.

[0011] According to a specific embodiment of the present invention, the chromatographic column used in the high performance liquid chromatography detection is selected from a C18 chromatographic column. @ C 18 (4.6mm×250mm, 5μm), PFchrom C 18 (4.6mm×250mm, 5μm), SinoChrom ODS-BP C 18 (4.6mm×250mm, 5μm), Welch Blossmate @ C 18 (4.6mm×250mm 5μm), Agilent 5TC C 18 (4.6mm×250mm 5μm) found that Welch Xtimate @ C 18 The column has a better separation effect on the characteristic spectrum of the pharmaceutical composition of the present invention.

[0012] According to a specific embodiment of the present invention, the pretreatment comprises: mixing the pharmaceutical composition with methanol and then performing ultrasonic treatment, supplementing the weight of the resulting mixed solution with methanol, shaking, filtering, collecting the filtrate, and obtaining a test solution. The inventors obtained the above-mentioned preferred pretreatment conditions through a large number of experiments, thereby effectively extracting the characteristic components corresponding to the common characteristic peaks with few impurities, thereby helping to obtain a characteristic spectrum with high separation and good peak shape.

[0013] According to a specific embodiment of the present invention, the ultrasonic treatment is carried out at a power of 220-280 W and a frequency of 35-45 kHz for 20-40 minutes, so as to fully extract the components of the pharmaceutical composition of the present invention.

[0014] According to a specific embodiment of the present invention, the concentration of methanol is 90-100% by volume, so as to fully extract the components of the pharmaceutical composition of the present invention.

[0015] According to a specific embodiment of the present invention, the method further comprises: under the same conditions, respectively subjecting the reference solution and the plurality of test solution to high performance liquid chromatography detection, preferably, the reference solution is selected from Solution, solution or a mixture of the two. As a reference, the resolution is good. Specifically, solution and The solution can be configured as two solutions separately or as the same mixed solution.

[0016] According to a specific embodiment of the present invention, the method for preparing the reference solution comprises: taking a reference substance An appropriate amount was accurately weighed and dissolved in methanol to prepare a solution containing about 0.1 mg of each substance per ml, which served as a reference solution.

[0017] According to a specific embodiment of the present invention, the number of the multiple different batches is at least 5 (eg, 5, 6, 7, 8, 9 or 10), so as to ensure the accuracy of the constructed characteristic spectrum.

[0018] According to a specific embodiment of the present invention, the characteristic spectrum includes 11 common characteristic peaks, with the seventh characteristic peak as the reference peak, and the ratios of the relative retention times of the 11 common characteristic peaks to the retention time of the reference peak are 0.40±10%, 0.55±10%, 0.63±10%, 0.66±10%, 0.87±10%, 0.97±10%, 1.00, 1.03±10%, 1.05±10%, 1.18±10%, and 1.26±10%, respectively.

[0019] refer to Figure 1 The characteristic spectrum of the pharmaceutical composition of the present invention includes 11 common characteristic peaks, and the common characteristic peaks are: the substance corresponding to peak No. 1 has the following structure: The substance corresponding to peak 2 has the following structure: Peak 3 is a common characteristic peak, and the substance corresponding to peak 4 has the following structure: Peak 5 is a common characteristic peak, peak 6 is a common characteristic peak, and the substance corresponding to peak 7 has the following structure:

[0020] The substance corresponding to peak 8 has the following structure: Peak No. 9 is a common characteristic peak, Peak No. 10 is a common characteristic peak, and Peak No. 11 is a common characteristic peak.

[0021] Preferably, the characteristic spectrum of the pharmaceutical composition includes 11 common characteristic peaks, among which peak 7 is the reference peak S; the relative retention time of each characteristic peak and the S peak is within ±10% of the specified value; the specified value is: 0.40 (peak 1), 0.55 (peak 2), 0.63 (peak 3), 0.66 (peak 4), 0.87 (peak 5), 0.97 (peak 6), 1.00 (peak 7, S), 1.03 (peak 8), 1.05 (peak 9), 1.18 (peak 10), 1.26 (peak 11).

[0022] In another aspect of the present invention, the present invention proposes a method for the pharmaceutical composition of the present invention, characterized in that it includes: pre-treating the sample to be tested to obtain a sample solution; performing high-performance liquid chromatography on the sample solution to obtain a chromatogram; analyzing the chromatogram with the characteristic spectrum obtained by the above method to determine whether the sample to be tested is the pharmaceutical composition of the present invention; the detection conditions of the high-performance liquid chromatography are as defined in the method for constructing the characteristic spectrum. Thus, the pharmaceutical composition of the present invention can be accurately identified by using the identification method according to the above invention.

[0023] According to a specific embodiment of the present invention, the pretreatment is as defined in the method for constructing the characteristic profile of the pharmaceutical composition described above.

[0024] According to a specific embodiment of the present invention, if the similarity between the chromatogram and the characteristic spectrum is not less than 0.9, the sample to be tested is identified as the pharmaceutical composition of the present invention; specifically, the similarity is obtained by using the fingerprint similarity evaluation software V2.0 of the "Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System" formulated by the State Pharmacopoeia Committee.

[0025] It should be noted that the features and advantages described above for the method for constructing the characteristic spectrum are also applicable to the method for identifying the pharmaceutical composition of the present invention and will not be described in detail here.

[0026] In another aspect of the present invention, the present invention proposes a method for detecting the pharmaceutical composition of the present invention, characterized in that it includes: configuring a known concentration solution of a characteristic component corresponding to at least one characteristic peak in the characteristic spectrum obtained by the method for constructing the characteristic spectrum as a reference solution; pretreating the sample to obtain a test solution; performing high performance liquid chromatography on the reference solution and the test solution to obtain a chromatogram; analyzing the chromatogram of the reference solution and the chromatogram of the test solution to determine the content of at least one characteristic component in the sample; wherein the high performance liquid chromatography detection conditions are as defined in the method for constructing the characteristic spectrum as described above. Thus, the method of the embodiment of the present invention can accurately determine the content of characteristic components in the pharmaceutical composition of the present invention, and has the advantages of good reproducibility, strong specificity, high precision and strong stability, and is suitable for popularization and application.

[0027] According to a specific embodiment of the present invention, the preprocessing is as defined in the method for constructing the characteristic map described above.

[0028] It should be noted that the features and advantages described above for the method for constructing the characteristic spectrum are also applicable to the method for detecting the pharmaceutical composition of the present invention and will not be described in detail here.

[0029] In another aspect of the present invention, the present invention proposes a method for quality control of the pharmaceutical composition of the present invention, characterized in that it includes: pre-treating a sample of the pharmaceutical composition to obtain a test solution; subjecting the test solution to high performance liquid chromatography to obtain a chromatogram; analyzing the chromatogram with the characteristic spectrum obtained by the method for constructing the characteristic spectrum described above to determine whether the quality of the sample to be tested meets the standard; wherein the high performance liquid chromatography detection conditions are as defined in the method for constructing the characteristic spectrum described above. Thus, the method according to an embodiment of the present invention can accurately determine whether the pharmaceutical composition of the present invention meets the requirements, thereby effectively controlling the quality.

[0030] According to a specific embodiment of the present invention, the preprocessing is as defined in the method for constructing the characteristic map described above.

[0031] According to a specific embodiment of the present invention, if the similarity between the chromatogram and the characteristic spectrum is not less than 0.9, the quality of the pharmaceutical composition is deemed to be up to standard.

[0032] It should be noted that the features and advantages described above for the method for constructing the characteristic spectrum of the pharmaceutical composition described in the present invention are also applicable to the method for quality control of the pharmaceutical composition described in the present invention and will not be repeated here.

[0033] In another aspect of the present invention, the present invention provides a pharmaceutical composition, characterized in that the pharmaceutical composition contains active ingredients, and the active ingredients are made from the following Chinese medicinal raw materials: honeysuckle, forsythia, Scrophularia, Isatis root, red peony root, Scutellaria baicalensis, mulberry leaf, chrysanthemum, Peucedanum chinense, bitter almond, burdock fruit, Alisma orientalis, Sterculia lychnophora, fried silkworm, cicada shell, and Oroxylum indicum; preferably, the weight ratio of any one raw material selected from honeysuckle, forsythia, Scrophularia ningpoensis, Isatis root to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, Peucedanum chinense, bitter almond, burdock fruit, Alisma orientalis, Sterculia lychnophora, silkworm, cicada shell, and Oroxylum indicum is 2.5:1, and the weight ratio of Scutellaria baicalensis to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, Peucedanum chinense, bitter almond, burdock fruit, Alisma orientalis, Sterculia lychnophora, silkworm, cicada shell, and Oroxylum indicum is 1:1. Specifically, in the pharmaceutical composition, the active ingredient is made of the following Chinese medicinal raw materials in the following weight percentages: based on the total weight of the Chinese medicinal raw materials, 11.112% honeysuckle, 11.112% forsythia, 11.112% scrophularia, 11.112% isatis root, 4.444% red peony root, 6.668% scutellaria, 4.444% mulberry leaf, 4.444% chrysanthemum, 4.444% peucedanum, 4.444% bitter almond, 4.444% arctium, 4.444% oriental water plantain, 4.444% sterculia lychnophora, 4.444% bombyx batryticatus, 4.444% cicada shell, and 4.444% oroxylum butterfly.

[0034] Specifically, the pharmaceutical composition has a characteristic spectrum constructed by the method of the first aspect of the present invention, which includes 11 characteristic peaks, and the substance corresponding to the first characteristic peak has the following structure: The substance corresponding to the second characteristic peak has the following structure: The substance corresponding to the fourth characteristic peak has the following structure: The fifth characteristic peak is a common characteristic peak, and the substance corresponding to the seventh characteristic peak has the following structure: The substance corresponding to the eighth characteristic peak has the following structure:

[0035] Among them, taking the seventh characteristic peak as the reference peak, the ratios of the relative retention times of the 11 characteristic peaks to the retention time of the reference peak were 0.40±10.0%, 0.55±10.0%, 0.63±10.0%, 0.66±10.0%, 0.87±10.0%, 0.97±10.0%, 1.00, 1.03±10.0%, 1.05±10.0%, 1.18±10.0%, and 1.26±10.0%, respectively.

[0036] Or, specifically, in the characteristic spectrum of the pharmaceutical composition of the present invention, the relative peak areas of the 11 characteristic peaks are 1.05, 0.46, 0.42, 1.798, 0.367, 0.222, 1, 0.332, 0.728, 0.173, and 0.212, respectively.

[0037] In another aspect of the present invention, the present invention also provides a method for preparing the pharmaceutical composition of the present invention, which comprises (or consists of the following):

[0038] (1) crushing all the Radix Isatidis and Bombyx Batryticatus, sieving and mixing;

[0039] (2) Grind all the honeysuckle, forsythia, red peony root and bitter almond, heat and reflux with ethanol, combine the extracts, filter and set aside the filtrate;

[0040] (3) boiling the residue obtained in step (2) and all the remaining ten medicinal materials (Scrophularia ningpoensis, Scutellaria baicalensis, Morus alba, Chrysanthemum, Peucedanum chinense, Arctium lappa, Rhizoma Alismatis, Sterculia lychnophora, Periostracum Cicadae, Oroxylum indicum) with water, filtering, concentrating the filtrate, adding ethanol, stirring, standing, filtering, combining the filtrate with the ethanol extract obtained in step (2), and concentrating;

[0041] (4) The powder obtained in step (1) and the thick paste obtained in step (3) are mixed and granulated.

[0042] According to a specific embodiment of the present invention, in the preparation method of the present invention, the granulation in step (4) is to directly perform boiling granulation on the mixed paste powder (i.e., the material obtained by mixing the powder obtained in step (1) and the thick paste obtained in step (3)) and water.

[0043] According to a specific embodiment of the present invention, in the preparation method of the present invention, the pulverizing in step (1) is to pulverize the medicinal material into fine powder, specifically, the fine powder is passed through a 100-120 mesh sieve.

[0044] According to a specific embodiment of the present invention, in the preparation method of the present invention, the pulverizing in step (2) is to pulverize the medicinal material into coarse powder, specifically, the coarse powder is passed through a 40-60 mesh sieve.

[0045] According to a specific embodiment of the present invention, in the preparation method of the present invention, the ethanol in step (2) is 60%-90% ethanol.

[0046] According to a specific embodiment of the present invention, in the preparation method of the present invention, the number of heating reflux in step (2) is 1-5 times, preferably 2-4 times; and / or the heating reflux time in step (2) is 1-5 hours / time, preferably 2-4 hours / time.

[0047] According to a specific embodiment of the present invention, in the preparation method of the present invention, the number of decoctions in step (3) is 1-5 times, preferably 2-4 times; and / or the decoction time in step (3) is 1-5 hours / time, preferably 2-4 hours / time.

[0048] According to a specific embodiment of the present invention, in the preparation method of the present invention, ethanol is added in step (3) to make the alcohol content reach 60%-80%.

[0049] According to a specific embodiment of the present invention, in the preparation method of the present invention, the filtrate in step (3) is concentrated to a thick paste with a relative density of 1.05-1.10 at a temperature of 60°C-80°C, preferably concentrated to a thick paste with a relative density of 1.05-1.10 at a temperature of 65°C-75°C.

[0050] According to a specific embodiment of the present invention, in the preparation method of the present invention, the concentration in step (3) is concentrated to a thick paste with a relative density of 1.38-1.40 at a temperature of 60°C-80°C, preferably concentrated to a thick paste with a relative density of 1.38-1.40 at a temperature of 65°C-75°C.

[0051] According to a specific embodiment of the present invention, in the preparation method of the present invention, ethanol can be recovered while concentrating in step (3).

[0052] In another aspect of the present invention, the present invention provides use of the pharmaceutical composition of the present invention in preparing a medicament for treating or preventing throat diseases such as acute pharyngitis and / or acute laryngitis.

[0053] Beneficial effects:

[0054] 1. The method for constructing the characteristic spectrum of the present invention overcomes the defect that the quality of the relevant dosage form cannot be fully detected from multiple angles due to the interference caused by the complex chemical composition of the pharmaceutical composition of the present invention; the present invention obtains 11 common characteristic peaks and realizes the effective separation of the 11 common characteristic peaks, with good peak shape and no interference, thereby realizing the quality detection of the effectiveness of the pharmaceutical composition of the present invention from multiple angles and in an all-round manner, and further realizing the quality control of the pharmaceutical composition of the present invention. The quality detection method is simple to operate, has good reproducibility and stability, reliable recovery rate, low detection cost and high detection efficiency after methodological verification, and the application of this method plays a vital role in the product quality stability of the pharmaceutical composition of the present invention.

[0055] 2. The present invention has established a set of rapid, comprehensive and highly specific quality detection methods for the pharmaceutical composition of the present invention.

[0056] 3. The characteristic spectrum constructed by the present invention can perform quality control on the pharmaceutical composition of the present invention from multiple angles and in all directions, providing more powerful guarantee for the authenticity identification and large-scale production of the pharmaceutical composition of the present invention.

[0057] 4. The present invention detects corresponding characteristic peaks in the characteristic spectra of different batches, and the similarity of the fingerprint spectra is greater than 0.9, indicating that the product quality is uniform and stable.

[0058] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0060] Figure 1 The chromatogram of the test sample using gradient elution condition 1 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0061] Figure 2 The chromatogram of the test sample using gradient elution condition 2 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0062] Figure 3 The chromatogram of the test sample using gradient elution condition 3 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0063] Figure 4 The chromatogram of the test sample using gradient elution condition 5 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0064] Figure 5 The chromatogram of the test sample using gradient elution condition 5 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0065] Figure 6 The chromatogram of the test sample using gradient elution condition 6 in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0066] Figure 7 The calibration results of the common characteristic peaks in the characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0067] Figure 8The chromatogram of the test sample for investigating the attribution of characteristic peaks in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown. The characteristic peaks in the figure are, from top to bottom, the test solution, the reference substance

[0068] of solution;

[0069] Fig. 9 The stability test results of the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0070] Fig.10 The results of the precision test in the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0071] Fig.11 The results of the repeatability test in the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0072] Fig.12 The intermediate precision Shimadzu-10AT VP test results of the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0073] Fig.13 The results of precision ThermoU3000 test in the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown;

[0074] Fig.14 The results of the reproducibility test in the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown in the laboratory 2Welch Xtimate @ C 18 Chromatographic columns;

[0075] Fig.15 The results of the reproducibility test in the methodological investigation experiment of the pharmaceutical composition according to the embodiment of the present invention are shown, laboratory 2 Agilent 588925-902 chromatography column;

[0076] Fig.16 The chromatogram of the test sample with the acetonitrile ratio reaching 70% in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention is shown;

[0077] Fig.17 It shows the detection wavelength investigation in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention - the chromatogram of the test sample detected by methanol ultrasound at a wavelength of 230nm;

[0078] Fig.18The chromatograms of the test samples extracted with methanol and 70% ethanol are shown in the investigation experiment of the HPLC characteristic spectrum of the pharmaceutical composition according to the embodiment of the present invention, in which the extraction solvents for preparing the test sample solution were investigated. DETAILED DESCRIPTION

[0079] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0080] 1. Preparation Example

[0081] Preparation Example 1

[0082] Formula:

[0083] 125 parts of Flos Honeysuckle, 125 parts of Fructus Forsythiae, 125 parts of Scrophulariae, 125 parts of Radix Isatidis, 50 parts of Red Peony Roots, 75 parts of Radix Scutellariae, 50 parts of Folium Mori, 50 parts of Flos Chrysanthemi, 50 parts of Peucedanum, 50 parts of Amarum Bitter, 50 parts of Fructus Arcti, 50 parts of Rhizoma Alismatis, 50 parts of Sterculia Lycopodiellae, 50 parts of Bombyx Batryticatus, 50 parts of Periostracum Cicadae, 50 parts of Oroxylum Indici.

[0084] The preparation method of the pharmaceutical composition is as follows:

[0085] (1) Grind all the Radix Isatidis and Bombyx Batryticatus into fine powder;

[0086] (2) Grind all the honeysuckle, forsythia, red peony root and bitter almond into coarse powder (40-60 mesh), extract twice with 80% ethanol under heating and reflux, each time for 2 hours, combine the extracts, filter and set aside the filtrate;

[0087] (3) boiling the residue obtained in step (2) and the remaining ten medicinal materials (Scrophularia ningpoensis, Scutellaria baicalensis, Morus alba, Chrysanthemum, Peucedanum chinense, Arctium lappa, Rhizoma Alismatis, Sterculia lychnophora, Periostracum Cicadae, Oroxylum indicum) in water twice, each time for 2 hours, combining the decoctions, filtering, concentrating the filtrate to a clear paste having a relative density of 1.05 to 1.10 at a temperature of 70° C., adding ethanol to make the alcohol content reach 70%, stirring evenly, standing, filtering, combining the filtrate with the above ethanol extract, recovering the ethanol and concentrating to a thick paste having a relative density of 1.38 to 1.40 at a temperature of 70° C.;

[0088] (4) The fine powder obtained in step (1) is mixed with the thick paste obtained in step (3), and then directly boiled with water to obtain granules, which can be loaded into capsules to obtain capsules.

[0089] 2. Experimental Examples

[0090] The test instruments, reagents, samples and reference substances used in the embodiments of the present invention are shown in Tables 1-5.

[0091] Table 1 Test instruments

[0092]

[0093] Table 2 Test reagents

[0094] name batch number level Manufacturer Methanol 20065185 HPLC TEDIA Acetonitrile 21101027G113 HPLC OCEANPAK Phosphoric acid 20210902 HPLC Tianjin Tianli Chemical Reagent Ultrapure water / First-class water self made

[0095] Table 3 Test samples

[0096]

[0097] Table 4 Test reference substances

[0098]

[0099]

[0100]

[0101] Table 5 Appearance of test samples

[0102]

[0103]

[0104] Example 1

[0105] Take 10 pellets of the contents of the 6 batches of samples for the test, prepare the test solution in the same way as the test solution, and perform the following operations in Laboratory 1:

[0106] (1) Chromatographic conditions: Octadecyl bonded silica gel as filler (Welch Xtimate @ C 18 , column length 25cm, inner diameter 4.6mm, particle size 5μm); 0.1 mass% phosphoric acid as mobile phase A, acetonitrile as mobile phase B, gradient elution according to the provisions in Table 6-11, balance for 10 minutes before injection; flow rate 1.0mL per minute; column temperature 30℃; detection wavelength 230nm. The resolution of the peak (also referred to herein as “reference substance 1”) from the adjacent chromatographic peaks should be greater than 1.5;

[0107] (2) Preparation of test solution: Grind the sample contents, take 0.5 g of the fine powder, accurately weigh it, place it in a stoppered conical flask, accurately add 25 mL of 100% methanol, weigh it, ultrasonically treat it (250 W, 40 kHz) for 30 minutes, let it cool to room temperature, weigh it again, make up the loss with 100% methanol, shake it well, filter it, and take the filtrate to obtain the solution.

[0108] (3) Preparation of reference solution: Take reference substance 1, An appropriate amount of (also referred to herein as "reference substance 2") was accurately weighed and dissolved in 100% methanol to prepare a solution containing approximately 0.1 mg of each per 1 mL, which served as a reference solution.

[0109] (4) Determination method: Accurately pipette 10 μL of reference solution and test solution respectively, inject into liquid chromatograph, measure, and record the chromatogram. Figure 1-6 .

[0110] Table 6 Gradient elution conditions 1

[0111]

[0112] Table 7 Gradient elution conditions 2

[0113]

[0114] Table 8 Gradient elution conditions 3

[0115]

[0116] Table 9 Gradient elution conditions 4

[0117]

[0118] Table 10 Gradient elution conditions 5

[0119]

[0120] Table 11 Gradient elution conditions 6

[0121]

[0122]

[0123] Conclusion: The gradient elution of mobile phase with different proportions in acetonitrile and 0.1 mass% phosphoric acid system was investigated. When the proportion of acetonitrile reached 60%, the main components in the prescription could be almost completely eluted out of the column. Finally, the chromatographic conditions to be verified were determined. The separation effect of each chromatographic peak in the test solution was good, and the baseline separation was basically achieved, which could meet the detection requirements.

[0124] Example 2 Determining the characteristic spectrum

[0125] The chromatograms of the test solutions of the six batches of samples used in the experiment were analyzed and compared, and the "Chinese Medicine Fingerprint Similarity Software" 2.0 was used for calibration to determine their common characteristic peaks.

[0126] Results: More than 20 chromatographic peaks were separated from the chromatograms of the 6 batches of test solution, of which there were 14 characteristic peaks in total, and the area of ​​the characteristic peaks accounted for more than 90% of the total peak area. The results are shown in Tables 12-14. Figure 7 .

[0127] Taking the peak of reference substance 1 as the reference peak, calculate the relative retention time and peak area ratio. The calculation formula is as follows:

[0128]

[0129] Table 12 Common characteristic peak matching results

[0130]

[0131]

[0132] Table 13 Relative retention time of common characteristic peaks

[0133]

[0134]

[0135] Table 14 Common characteristic peak relative peak area

[0136]

[0137] 1. Attribution of characteristic peaks

[0138] Positioning reference solution: Take reference 1, reference 2, (also referred to herein as "reference substance 3") (also referred to herein as "reference substance 4"), (also referred to herein as "reference substance 5"), (also referred to herein as "reference substance 6")

[0139] (also referred to herein as "Control 7"),

[0140] An appropriate amount of (also referred to herein as "reference substance 8") was accurately weighed and dissolved in 100% methanol to prepare a solution containing approximately 0.1 mg of each per 1 mL, which served as the positioning reference solution.

[0141] The sample was injected and measured under the above chromatographic conditions to be verified, and the peaks were compared with those in the chromatogram of the test solution (batch number: JD121090082) to attribute the characteristic peaks. The results are shown in Table 15. Figure 8 .

[0142] Table 15 Characteristic peak attribution

[0143]

[0144]

[0145] Results: Seven known components were identified by characteristic peak identification in the chromatogram of the test solution, among which the peaks of reference substance 1, reference substance 3, reference substance 4, reference substance 2 and reference substance 7 were relatively obvious and could be used as characteristic known peaks; reference substance 6 and reference substance 5 were detected weakly and were not used as characteristic peaks; the absorption of reference substance 8 was weak at 230nm, and the chromatographic peak of reference substance 8 was not reflected in this characteristic spectrum.

[0146] 2. Investigation of reference solution

[0147] The peaks of known components detected in the test solution are more obvious, with reference 1, reference 2 and reference 4 located in the middle of the characteristic spectrum. Reference 1 and reference 2 can better reflect the separation effect of the spectrum, and therefore they are used as references.

[0148] Preparation of reference solution Take appropriate amount of reference substance 1 and reference substance 2, weigh accurately, and dissolve in methanol to make a solution containing approximately 0.1 mg of each per 1 mL, which is used as the reference solution.

[0149] 3. Methodological investigation

[0150] With reference to the General Rules 9101 "Guidelines for Validation of Analytical Methods" of the 2020 edition of the Chinese Pharmacopoeia and the Technical Requirements for Fingerprint Studies of Traditional Chinese Medicine Injections, characteristic spectrum / fingerprint methodology investigations were conducted, mainly including solution stability, precision, repeatability, intermediate precision, durability and reproducibility tests. The acceptable standards for validation are mainly based on the acceptable range of the content and precision of the components to be tested in the samples of the Technical Requirements for Fingerprint Studies of Traditional Chinese Medicine Injections and the General Rules 9101 "Guidelines for Validation of Analytical Methods" of the 2020 edition of the Chinese Pharmacopoeia.

[0151] (1) Stability test

[0152] Prepare the test solution (batch number: JD121090082), accurately pipette 10 μL of the test solution, place it at room temperature and protected from light for 0, 4, 8, 12, 28, and 48 h, respectively, and measure the relative retention time and peak area ratio of the common characteristic peaks. At the same time, perform similarity evaluation using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" V2.0 version to investigate the stability of the test solution.

[0153] Verification requirements: The relative retention time RSD of the characteristic peak in the chromatogram of the test solution is ≤2.0%, the relative peak area RSD is ≤5.0%, and the similarity is >0.9. The test solution can be considered to be stable.

[0154] Verification results: In the test solution, the RSD of the relative retention time of each characteristic peak is ≤1.34%, and the RSD of the relative peak area is ≤3.42% (except for peaks 7, 10, 11, and 17). Peaks 7, 10, 11, and 17 do not show obvious degradation trends, mainly because they account for fewer total peaks and the separation effect affects the integration results. The similarities are all greater than 0.9, indicating that the test solution is stable within 48 hours. The results are shown in Tables 16-17. Fig. 9 , where S1-S6 represent characteristic spectra at different times respectively.

[0155] Table 16 Solution stability similarity evaluation

[0156]

[0157]

[0158] Table 17 Stability test results

[0159]

[0160]

[0161] (2) Precision test

[0162] Prepare the test solution (batch number: JD121090082), draw 10 μL of the test solution, and inject it 6 times continuously. Calculate the relative retention time and relative peak area of ​​the common characteristic peaks in the characteristic spectrum. At the same time, perform similarity evaluation using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" V2.0 version to examine the precision of the instrument.

[0163] Verification requirements: The relative retention time RSD of the characteristic peaks in the chromatogram of the test solution is ≤2.0%, the relative peak area RSD is ≤5.0%, and the similarity is >0.9.

[0164] Verification results: 6 test results, relative retention time RSD ≤ 1.1%, relative peak area RSD ≤ 3.19% (except for peak 10), similarity is greater than 0.9, indicating that the instrument has good precision. The results are shown in Table 18-19. Fig.10 .

[0165] Table 18 Precision Similarity Evaluation

[0166]

[0167] Table 19 Precision test results

[0168]

[0169]

[0170] (3) Repeatability test

[0171] Six samples of the test solution (batch number: JD121090082) were prepared, and 10 μL of each solution was injected for analysis. The relative retention time and relative peak area of ​​the common characteristic peaks in the characteristic spectrum were calculated. At the same time, the similarity evaluation was performed using the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" V2.0 version to examine the reproducibility of the experimental method.

[0172] Verification requirements: The relative retention time RSD of the common characteristic peaks in the chromatogram of the test solution is ≤2.0%, the relative peak area RSD is ≤15.0%, and the similarity is >0.9.

[0173] Verification results: 6 test results, relative retention time RSD ≤ 1.88%, relative peak area RSD ≤ 10.49% (except for peak 13), the deviation of peak 13 is mainly due to the difference in integration caused by the separation effect, similarity > 0.9, indicating that the repeatability of the determination method is good. The results are shown in Table 20-21. Fig.11 .

[0174] Table 20 Repeatability similarity evaluation

[0175]

[0176] Table 21 Repeatability test results

[0177]

[0178]

[0179] (4) Intermediate precision

[0180] Different personnel took the contents of this product (batch number: JD121090082) to prepare the test solution according to the preparation method of the test solution, and took 10 μL of each sample and injected it into Shimadzu-10AT VP and Thermo U3000 high performance liquid chromatograph for analysis, and calculated the relative retention time of the characteristic peaks in the fingerprint. At the same time, the "Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine" V2.0 version was used to conduct a repeatability test (batch number: JC121030022) to compare the spectra for similarity evaluation, and to examine the intermediate precision of the experimental method.

[0181] Verification requirements: In the characteristic spectrum, the relative deviation of the characteristic peak relative retention time is specified to be within 5% and the similarity is greater than 0.9.

[0182] Verification results: Different personnel and different instruments determined that the common characteristic peaks were all detected, the RSD of the relative retention time was ≤4.36%, and the similarity was >0.9, indicating that the intermediate precision of the determination method was good. The results are shown in Table 22-23. Figure 12-13 .

[0183] Table 22 Intermediate precision similarity evaluation (Mark peak)

[0184]

[0185] Table 23 Intermediate precision test results

[0186]

[0187]

[0188] (5) Reproducibility test

[0189] The test samples of this product (batch number: JD121090082) were measured in different laboratories, and characteristic spectra were drawn to examine the reproducibility of the method.

[0190] Verification requirements: The relative retention time of the characteristic peaks in the characteristic spectrum is within the specified range and the separation meets the requirements.

[0191] Verification results: Laboratory 2 used (Welch Xtimate @ C 18, 4.6mm×250mm, 5μm) and Agilent588925-902, 4.6mm×250mm, 5μm). The characteristic spectrum of the sample is consistent with the standard characteristic spectrum of laboratory 1. Although the separation of peaks 2 and 7 did not completely achieve baseline separation, it basically met the separation requirements; the relative retention time of the characteristic peaks was not controlled within ±8% except for peaks 1 and 8, and the other peaks fully met the requirements, indicating that the method reproducibility is feasible and the durability meets the requirements; the analysis effects of peaks 6 and 7 on different chromatographic columns and instruments are quite different, and it is recommended not to be listed as characteristic peaks. The results are shown in Table 24. Figure 14-15 .

[0192] Table 24 Separation characteristic spectrum results of laboratory 2

[0193]

[0194]

[0195] The common characteristic peaks of 6 batches of samples used in the experiment were analyzed. Among them, peaks 2, 3, 4, 6, 7, 11, and 13 were not used as common characteristic peaks because they accounted for a small proportion of the total peaks, were unstable, and had poor separation effects. Peaks 1, 5, 8, 9, 10, 12, 14, 15, 16, 17, 18, 19, 20, and 21 accounted for 2% to 14% of the total peaks and had better chromatographic separation, so they were planned to be used as common characteristic peaks.

[0196] Referring to the results of the intermediate precision and reproducibility experiments, the separation effects of peaks 12 (0.74) and 14 (0.80) on different chromatographic columns were quite different. Analysis showed that they were combination peaks of different substances, not single component chromatographic peaks, with poor stability and repeatability. Therefore, they were not included in the characteristic peaks. Peak 1 (0.17) had poor stability in different laboratories and was not listed as a characteristic peak.

[0197] 4. Proposed feature map

[0198] There should be 11 characteristic peaks in the characteristic spectrum of the test sample. The seventh characteristic peak should be taken as the reference peak (S) and the relative retention time of each characteristic peak and the S peak should be calculated.

[0199] The relative retention times of the 11 characteristic peaks of this product are: 0.40 (peak 1), 0.55 (peak 2), 0.63 (peak 3), 0.66 (peak 4), 0.87 (peak 5), 0.97 (peak 6), 1.00 (peak 7, S), 1.03 (peak 8), 1.05 (peak 9), 1.18 (peak 10), 1.26 (peak 11), of which peak 1 is Peak 2 Peak 4 Peak 7 (S) is Peak 8

[0200] 5. Revision scope of characteristic spectrum limit

[0201] Comparing the relative retention times of the 11 characteristic peaks, the deviations of the measurements by different instruments were within ±5% of the mean, and the measurements by different laboratories were within ±9% of the above specified values. Therefore, the relative retention time of each characteristic peak was determined to be within ±10.0% of the specified value, see Figure 6 .

[0202] Based on the above results, 11 characteristic peaks should be present in the characteristic spectrum of the test sample. The seventh characteristic peak is taken as the reference peak (S) peak. The relative retention time of each characteristic peak and the S peak is calculated, which should be within ±10.0% of the specified value. The specified value is:

[0203] 0.40 (peak 1), 0.55 (peak 2), 0.63 (peak 3), 0.66 (peak 4), 0.87 (peak 5), 0.97 (peak 6), 1.00 (peak 7, S), 1.03 (peak 8), 1.05 (peak 9), 1.18 (peak 10), 1.26 (peak 11), of which peak 1 is Peak 2 Peak 4 Peak 7 (S) is Peak 8

[0204] 6. Similarity evaluation

[0205] The characteristic spectrum test data of 6 batches (JD121090082, JD121090092, JD121090102, JD121090112, JD121090122, JD121090132) were imported into the Chinese medicine fingerprint spectrum similarity software, and the control characteristic spectrum of the characteristic spectrum was obtained by calculation by the Chinese medicine fingerprint spectrum similarity software (see Figure 6 ), each characteristic spectrum test data is calculated by MARK peak (characteristic peak) similarity, and the similarity of each batch of samples is compared.

[0206] The results showed that characteristic peaks were detected in the fingerprints of samples from different batches, and the similarity of MARK peaks (characteristic peaks) was ≥ 0.9, with no significant differences. The results are shown in Table 25.

[0207] Table 25 Fingerprint similarity evaluation results of different batches of samples

[0208]

[0209]

[0210] 7. System suitability investigation

[0211] The system suitability was investigated using the separation effect of the reference peak and adjacent peaks in the chromatogram of the test solution as the evaluation index. The results are shown in Table 26 below.

[0212] Table 26 System suitability investigation results

[0213]

[0214] Example 3

[0215] In this embodiment, the characteristic map is constructed according to the following method:

[0216] (1) Chromatographic conditions: Octadecyl bonded silica gel was used as the filler and the chromatographic column was selected from Welch Xtimate @ C 18 , column length 25cm, inner diameter 4.6mm, particle size 5μm; 0.05 mass% phosphoric acid as mobile phase A, acetonitrile as mobile phase B, gradient elution as specified in the table below, balance for 10 minutes before injection; flow rate 0.8mL per minute; column temperature 25℃; detection wavelength 200nm. The resolution between the reference peak and the adjacent chromatographic peak should be greater than 1.5;

[0217] Table 27 Elution conditions

[0218] Time (minutes) 0.05 mass% phosphoric acid (%) Acetonitrile (%) 0 90 10 3 90 10 7 85 15 35 70 30 52 50 50 60 40 60

[0219] (2) Preparation of test solution: Grind the sample contents, take 0.5 g of the fine powder, accurately weigh it, place it in a stoppered conical flask, accurately add 25 mL of 90% methanol, weigh it, ultrasonically treat it (220 W, 35 kHz) for 20 min, let it cool to room temperature, weigh it again, make up the loss with 90% methanol, shake it well, filter it, and take the filtrate to obtain the solution.

[0220] (3) Preparation and determination of reference solution: Same as in Example 1, recording the chromatogram.

[0221] The chromatogram of the test sample showed 11 common characteristic peaks. Taking the seventh characteristic peak as the reference peak, the ratios of the relative retention time of the 11 common characteristic peaks to the retention time of the reference peak were 0.40±10%, 0.55±10%, 0.63±10%, 0.66±10%, 0.87±10%, 0.97±10%, 1.00, 1.03±10%, 1.05±10%, 1.18±10%, and 1.26±10%. The separation of adjacent chromatographic peaks was greater than 1.5, and baseline separation was achieved between the characteristic peaks, which met the requirements.

[0222] Example 4

[0223] In this embodiment, the characteristic map is constructed according to the following method:

[0224] 1. Chromatographic conditions: Octadecyl bonded silica gel is used as a filler, and the chromatographic column is selected from Welch Xtimate@C18, with a column length of 25 cm, an inner diameter of 4.6 mm, and a particle size of 5 μm; 0.1 mass % phosphoric acid is used as mobile phase A, acetonitrile is used as mobile phase B, and gradient elution is performed (elution conditions are the same as in Example 1), and equilibration is performed for 10 minutes before injection; the flow rate is 1.0 mL per minute; the column temperature is 30° C.; the detection wavelength is 320 nm; the theoretical plate number calculated based on the reference peak should be not less than 10,000;

[0225] 2. Preparation and determination of test solution and reference solution: Same as Example 1, the characteristic spectrum obtained is shown in Fig.17 .

[0226] Conclusion: Although the peak shape at 320nm is more prominent and the baseline drift is smaller, the characteristic peak at 230nm is more obvious, which basically reflects the fingerprint characteristics of the sample. Therefore, the detection wavelength is determined to be 230nm.

[0227] Example 5

[0228] In this embodiment, the characteristic map is constructed according to the following method:

[0229] 1. Chromatographic conditions: same as in Example 1.

[0230] 2. Preparation of test solution: Take the sample content, grind it into powder, take 0.5g of fine powder, weigh it accurately, put it in a stoppered conical flask, add 25mL of 70% ethanol accurately, weigh it, ultrasonically treat it (250W, 40KHz) for 30 minutes, let it cool to room temperature, weigh it again, make up the lost weight with 70% ethanol, shake it well, filter it, and take the filtrate to obtain the solution;

[0231] 3. Preparation and determination of reference solution: Same as Example 1. The characteristic spectrum obtained is shown in Fig.18 .

[0232] The test sample solutions were prepared by direct extraction with methanol and 70% ethanol. The chromatographic peaks in the chromatograms of the test samples with different solvents were the same, the characteristic peaks were obvious, and the separation effects could meet the separation requirements. The methanol extract was easier to filter than the 70% ethanol extract (containing water). The experiment determined that methanol was used to prepare the test sample solution by ultrasonic treatment, so methanol was determined to be the extraction solvent.

[0233] summary

[0234] The characteristic spectrum / fingerprint spectrum developed by this method has certain specificity. The characteristic peaks basically reflect the main components with ultraviolet absorption and known effective ingredients in the pharmaceutical composition. The identification of 11 characteristic peaks better reflects the material basis of the pharmaceutical composition. The method has good durability and reproducibility and can be used as a basis for quality control of the pharmaceutical composition, for evaluating the uniformity, excellence and stability of the preparation quality, and for comparing the consistency of product quality before and after the change.

[0235] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A method for constructing a characteristic spectrum of a pharmaceutical composition, characterized in that: include: Providing a plurality of different batches of pharmaceutical composition samples; Under the same conditions, each sample was pretreated separately to obtain multiple test sample solutions; Under the same conditions, a plurality of the test sample solutions are subjected to high performance liquid chromatography to obtain a plurality of chromatograms; By comparing and analyzing a plurality of the chromatograms, common characteristic peaks are determined to obtain a characteristic spectrum of the pharmaceutical composition; Wherein, the mobile phase used in the high performance liquid chromatography detection includes: Mobile phase A, wherein the mobile phase A is a 0.05-0.15 mass % phosphoric acid solution; Mobile phase B, wherein the mobile phase B is acetonitrile; The elution mode of the mobile phase is gradient elution; Wherein, the pharmaceutical composition contains active ingredients, and the active ingredients are made from the following Chinese medicinal raw materials: honeysuckle, forsythia, Scrophularia, Isatis root, red peony root, scutellaria, mulberry leaf, chrysanthemum, Peucedanum peucedanum, bitter almond, burdock fruit, oriental rhizome, Sterculia lychnophora, fried silkworm, cicada shell, and oroxylum butterfly; preferably, the weight ratio of any one raw material selected from honeysuckle, forsythia, Scrophularia ningpoensis, and Isatis root to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, Peucedanum peucedanum, bitter almond, burdock fruit, oriental rhizome, Sterculia lychnophora, silkworm, cicada shell, and oroxylum butterfly is 2.5:1, and the weight ratio of Scutellaria baicalensis to any one raw material selected from red peony root, mulberry leaf, chrysanthemum, Peucedanum peucedanum, bitter almond, burdock fruit, oriental rhizome, Sterculia lychnophora, silkworm, cicada shell, and oroxylum butterfly is 1:

1.

2. The method according to claim 1, characterized in that: The elution mode of the mobile phase is as follows: 。 3. The method according to claim 1, characterized in that The mobile phase used in the HPLC detection has a flow rate of 0.8 to 1.2 mL / min, a column temperature of 25 to 35° C., and a wavelength of 200 to 250 nm; and / or, the chromatographic column is selected from a C18 chromatographic column, preferably a Welch Xtimate @ C 18 column.

4. The method according to claim 1, characterized in that: The pre-processing comprises: The pharmaceutical composition and methanol are mixed and then ultrasonically treated, the resulting mixture is supplemented with methanol to make up the weight, shaken well, filtered, and the filtrate is collected to obtain a test solution; Optionally, the ultrasonic treatment is carried out at a power of 220 to 280 W and a frequency of 35 to 45 kHz for 20 to 40 minutes; Optionally, the concentration of the methanol is 90 to 100 volume %.

5. The method according to claim 1, characterized in that Further including: Under the same conditions, the reference solution and the plurality of test solution are subjected to high performance liquid chromatography detection, preferably, the reference solution is selected Solution, solution or a mixture of the two; Optionally, the number of batches of the plurality of different batches is at least 5; Optionally, the characteristic spectrum includes 11 common characteristic peaks, among which the seventh characteristic peak is used as a reference peak, and the ratios of the relative retention times of the 11 common characteristic peaks to the retention time of the reference peak are 0.40±10%, 0.55±10%, 0.63±10%, 0.66±10%, 0.87±10%, 0.97±10%, 1.00, 1.03±10%, 1.05±10%, 1.18±10%, and 1.26±10%, respectively.

6. A method for identifying the pharmaceutical composition defined in claim 1, characterized in that: include: Pre-treating the sample to be tested to obtain a sample solution; Performing high performance liquid chromatography on the sample solution to obtain a chromatogram; Analyzing the chromatogram with the characteristic spectrum obtained by the method according to any one of claims 1 to 5 to determine whether the sample to be tested is the pharmaceutical composition defined in claim 1; The detection conditions of the HPLC are as defined in the method for constructing the characteristic spectrum of the pharmaceutical composition as claimed in any one of claims 1 to 3 and 5.

7. The method according to claim 6, characterized in that The pretreatment is as defined in the method of claim 4; If the similarity between the chromatogram and the characteristic spectrum is not less than 0.9, the sample to be tested is determined to be the pharmaceutical composition defined in claim 1.

8. A method for detecting the pharmaceutical composition defined in claim 1, characterized in that: include: A solution of a characteristic component of known concentration corresponding to at least one characteristic peak in the characteristic spectrum obtained by the method for constructing a characteristic spectrum of a pharmaceutical composition according to any one of claims 1 to 5 is prepared as a reference solution; Pre-treating the sample to obtain a test solution; The reference solution and the test solution are respectively subjected to high performance liquid chromatography to obtain a chromatogram; Analyzing the chromatogram of the reference solution and the chromatogram of the test solution to determine the content of at least one characteristic component in the sample; Wherein, the HPLC detection conditions are defined in the method for constructing the characteristic spectrum of the pharmaceutical composition as described in any one of claims 1-3 and 5.

9. The method according to claim 8, characterized in that The pre-treatment is as defined in the method of claim 4.

10. A method for quality control of the pharmaceutical composition defined in claim 1, characterized in that: include: Pretreating the pharmaceutical composition sample to obtain a test solution; The test solution is subjected to high performance liquid chromatography to obtain a chromatogram; Analyze the chromatogram with the characteristic spectrum obtained by the method according to any one of claims 1 to 5 to determine whether the quality of the sample to be tested meets the standard; Wherein, the HPLC detection conditions are as defined in the method according to any one of claims 1 to 3 and 5; Optionally, the pretreatment is as defined in the method of claim 4; Optionally, if the similarity between the chromatogram and the characteristic spectrum is not less than 0.9, the quality of the pharmaceutical composition is deemed to be up to standard.

11. The pharmaceutical composition according to claim 1, characterized in that The similarity between the high performance liquid chromatogram of the pharmaceutical composition and the characteristic spectrum constructed by the method of any one of claims 1 to 5 is not less than 0.

9.

12. Use of the pharmaceutical composition as defined in claim 11 in the preparation of a medicament for treating or preventing throat diseases such as acute pharyngitis and / or acute laryngitis.

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