Fingerprint spectrum of anti-asthma tablets and its establishment method

Abstract

This invention relates to the field of traditional Chinese medicine component analysis technology, specifically to the fingerprint spectrum of Juanxiao tablets and its establishment method. The method for establishing the fingerprint spectrum of Juanxiao tablets includes: using isoharonidin, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, nobiletin, and citrus reticulataein as reference standards, and Juanxiao tablets as the test sample; using HPLC to detect the reference standards and the test sample, and then comparing and analyzing them to determine the common characteristic peaks of Juanxiao tablets; wherein the HPLC conditions include: a C18 column, mobile phase A as a nitrile solvent, mobile phase B as a 0.05-0.15% phosphoric acid solution, and gradient elution. This method can effectively detect characteristic peaks, reduce the drying of impurity peaks, and the obtained fingerprint spectrum has high resolution and good peak shape. All characteristic peaks are baseline separated, enabling a more comprehensive and accurate evaluation of the quality of Juanxiao tablets.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine component analysis technology, and more specifically, to the fingerprint spectrum of Juanxiao tablets and its establishment method. Background Technology

[0002] Juanxiao tablets, as a representative formula for treating phlegm and blood stasis, are designed to address phlegm, blood stasis, and qi stagnation. The formula contains: Lepidium seed to purge lung congestion, relieve asthma, and promote diuresis and reduce swelling; Vitex fruit to dispel wind, release the exterior, stop coughing, and relieve asthma; Citrus reticulata peel to regulate liver qi; Citrus reticulata peel to invigorate spleen qi; Areca catechu to lower qi and eliminate stagnation. These three herbs work together to regulate qi flow, lower qi, and relieve asthma; Rhubarb to promote bowel movement and remove blood stasis; and Ginger to dispel cold, warm the stomach, and harmonize the middle jiao. With the goal of regulating qi flow, eliminating phlegm, and dispersing blood stasis, it embodies the effects of purging lung qi, regulating liver qi, invigorating spleen qi, and clearing bowel movement qi. It is used for acute exacerbations of bronchial asthma with fever, asthma, and phlegm stasis lurking in the lungs, characterized by coarse breathing, profuse phlegm, wheezing, paroxysmal coughing, and thick yellow phlegm.

[0003] Traditional Chinese medicine (TCM) fingerprinting refers to the chromatograms or spectra obtained by appropriate analysis of certain TCM materials or preparations after proper processing. It identifies the chemical characteristics of these materials and preparations, and is based on systematic research into the chemical components of TCM. It is primarily used to evaluate the authenticity, quality, and stability of TCM materials and semi-finished TCM preparations. It is characterized by both "holistic" and "fuzzy" properties. Fingerprinting is currently an internationally recognized quality control method capable of holistic component analysis, and it is of great significance for controlling the quality of TCM materials, extracts, and their preparations. The 2010 edition of the Chinese Pharmacopoeia included fingerprinting technology as a qualitative testing item for the first time.

[0004] Due to the complexity of traditional Chinese medicine components, the determination of fingerprint spectra of traditional Chinese medicine faces problems such as difficulty in separating characteristic peaks, large interference from impurity peaks, complex operation, and poor accuracy, precision, stability, and repeatability. Based on the above problems, the existing technology does not disclose a method for establishing the fingerprint spectra of Juanxiao tablets.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The purpose of this invention is to provide a fingerprint spectrum of anti-asthma tablets and a method for establishing it. The method provided by the embodiments of this invention can effectively detect characteristic peaks, reduce the drying of impurity peaks, and the obtained fingerprint spectrum has high separation and good peak shape. All characteristic peaks have achieved baseline separation, which can more comprehensively and accurately evaluate the quality of anti-asthma tablets.

[0007] This invention is implemented as follows:

[0008] In a first aspect, the present invention provides a method for establishing a fingerprint spectrum of Juanxiao tablets, comprising: using isoharmonin, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, norihesperidin and tangeretin as reference standards, and Juanxiao tablets as the test sample.

[0009] The reference standard and the test sample were detected by HPLC and then compared and analyzed to determine the common characteristic peaks of the anti-asthmatic tablets;

[0010] The HPLC conditions included: a C18 column, mobile phase A being a nitrile solvent, mobile phase B being a 0.05-0.15% phosphoric acid solution, and gradient elution.

[0011] In an optional embodiment, the nitrile solvent is selected from C2-C5 nitrile solvents, preferably C2-C3 nitrile solvents, and most preferably acetonitrile.

[0012] In an optional embodiment, the mobile phase B is a 0.1% phosphoric acid solution.

[0013] In an optional implementation, the HPLC conditions also include: a detection wavelength of 210-330 nm; a column temperature of 28-32 °C; and an injection volume of 5-10 μL.

[0014] In an optional implementation, the gradient elution process is as follows:

[0015]

[0016]

[0017] In an optional implementation, the method further includes: processing the test sample and the reference sample separately to form a test sample solution and a reference sample solution before HPLC detection.

[0018] In an optional embodiment, the step of forming the test solution includes: dissolving the anti-asthmatic tablets using an alcohol solvent, and subjecting the dissolution process to ultrasonic treatment;

[0019] Preferably, the process includes: adding 100-150 ml of alcohol solvent to each gram of the anti-asthmatic tablet, mixing and ultrasonicating, cooling after ultrasonication and reweighting to the initial weight, and then filtering.

[0020] In an optional embodiment, the step of forming the reference solution includes dissolving the reference standard separately using an alcohol solvent.

[0021] In an optional embodiment, the ingredients of the anti-asthmatic tablets are as follows, by weight: 418 parts of Lepidium apetalum, 418 parts of Citrus reticulata peel, 418 parts of Citrus reticulata peel, 625 parts of Vitex negundo fruit, 418 parts of Areca catechu, 125 parts of Rheum palmatum, and 100 parts of Zingiber officinale.

[0022] In an optional implementation, the method for forming the asthma tablet is as follows:

[0023] Ginger is juiced to make ginger juice; rhubarb is crushed to make rhubarb powder;

[0024] The remaining raw materials are distilled using steam, and the distillate is collected.

[0025] The distilled water is filtered to form the first filtrate.

[0026] The residue after distillation is extracted with water and then filtered to form a second filtrate.

[0027] The first filtrate and the second filtrate are mixed and then mixed with an alcohol solvent and allowed to stand. The mixture is then filtered to form a third filtrate, and the third filtrate is concentrated to form an extract powder.

[0028] The extract powder is mixed with rhubarb powder and then pulverized. After adding fillers and humectants, it is granulated and dried. Then, ginger juice and distillate are added and mixed evenly, and then fillers are added and tableted.

[0029] Secondly, embodiments of the present invention provide a fingerprint spectrum for an asthma-relieving tablet, which is obtained by the above-described method for establishing an asthma-relieving tablet fingerprint spectrum.

[0030] This invention offers the following advantages: By selecting specific reference standards and specific HPLC conditions, the embodiments of this invention solve the problems of difficult separation of characteristic peaks and significant interference from impurity peaks. The resulting fingerprint chromatogram exhibits high separation and good peak shape, with baseline separation achieved for each characteristic peak, enabling a more comprehensive and accurate evaluation of the quality of Juanxiao tablets. Furthermore, this method is simple to operate, accurate, and reliable, exhibiting good precision, stability, and repeatability. It overcomes the shortcomings of existing technologies that rely on single detection indicators and cannot reflect the intrinsic quality of Juanxiao tablet formulation process control, achieving a comprehensive and accurate evaluation of formulation quality, thereby more effectively ensuring formulation quality. When using similarity for evaluation, its standard fingerprint chromatogram can be used to comprehensively evaluate and control the quality of Juanxiao tablets, thus ensuring product quality stability and the safety and effectiveness of clinical use. Attached Figure Description

[0031] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0032] Figure 1 The HPLC chromatograms of 10 batches of anti-asthma tablets provided in Example 1 of the present invention;

[0033] Figure 2 This is a reference HPLC chromatogram of the anti-asthmatic tablets provided in Example 1 of the present invention;

[0034] Figure 3The HPLC chromatogram of the anti-asthma tablets provided in Example 2 of this invention;

[0035] Figure 4 The HPLC chromatogram of the anti-asthma tablets provided in Example 3 of this invention;

[0036] Figure 5 The HPLC chromatogram of the anti-asthma tablets provided in Example 4 of this invention;

[0037] Figure 6 The HPLC chromatogram of the anti-asthma tablets provided in Example 5 of this invention;

[0038] Figure 7 HPLC chromatograms provided as verification examples of the present invention;

[0039] Figure 8 The repeatability HPLC chromatograms provided for the verification examples of this invention;

[0040] Figure 9 HPLC chromatograms of stability provided for verification examples of this invention;

[0041] Figure 10 The HPLC chromatogram of the anti-asthmatic tablets provided in Comparative Example 1 of this invention is shown below.

[0042] Figure 11 The HPLC chromatogram of the anti-asthma tablets provided in Comparative Example 2 of this invention;

[0043] Figure 12 The HPLC chromatogram of the anti-asthmatic tablets provided in Comparative Example 3 of this invention is shown below.

[0044] Figure 13 The HPLC chromatogram of the anti-asthmatic tablets provided in Comparative Example 4 of this invention is shown below.

[0045] Figure 14 The HPLC chromatogram of the anti-asthmatic tablets provided in Comparative Example 5 of this invention is shown below.

[0046] Figure 15 This is the HPLC chromatogram of the anti-asthmatic tablets provided in Comparative Example 6 of the present invention. Detailed Implementation

[0047] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0048] In a first aspect, the present invention provides a method for establishing a fingerprint spectrum of asthma tablets, comprising:

[0049] Using isoharonin, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, nodosum glycoside, and citrus peel as reference standards, this invention uses hesperidin as a reference peak and identifies nine common peaks as the common peaks constituting the fingerprint spectrum of Juanxiao tablets, making the composition of the fingerprint spectrum richer and more comprehensive. Isoharonin, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, nodosum glycoside, and citrus peel were identified from the common peaks, further clarifying the chemical basis of Juanxiao tablets.

[0050] The reference solutions were prepared by dissolving the substances in alcohol solvents, such as C1-C5 monohydric alcohols or methanol.

[0051] The test sample is a tablet for treating asthma. The tablet is dissolved in an alcohol solvent, and the dissolution process is performed by sonication. Specifically, 100-150 ml of alcohol solvent is added for each gram of the tablet, and the initial weight is recorded after mixing. Then, the mixture is dissolved by sonication. During the sonication process, the mixed volume may be heated and some solvent may evaporate. Therefore, after the ultrasonic dissolution is completed, the mixture is allowed to cool to room temperature under natural conditions, and the solvent is added back to the initial weight. The mixture is then filtered to obtain the test solution.

[0052] The test solution can be used directly for injection without further dilution. The alcohol solvent can be a C1-C5 monohydric alcohol, such as methanol.

[0053] The samples for the pilot and trial production of Juanxiao tablets are as follows. The ingredients of Juanxiao tablets are as follows: 418 parts of Lepidium apetalum, 418 parts of Citrus reticulata peel, 418 parts of Citrus reticulata peel, 625 parts of Vitex negundo fruit, 418 parts of Areca catechu, 125 parts of Rheum palmatum, and 100 parts of Zingiber officinale.

[0054] The method for forming the aforementioned anti-asthma tablet is as follows:

[0055] Ginger is juiced to make ginger juice; rhubarb is crushed to make rhubarb powder;

[0056] The remaining raw materials are distilled using steam, and the distillate is collected.

[0057] The distilled water is filtered to form the first filtrate.

[0058] The residue after distillation is extracted with water and then filtered to form a second filtrate.

[0059] The first filtrate and the second filtrate are mixed and then mixed with an alcohol solvent and allowed to stand. The mixture is then filtered to form a third filtrate, and the third filtrate is concentrated to form an extract powder.

[0060] The extract powder is mixed with rhubarb powder and then pulverized. A filler, such as starch, and a wetting agent, such as ethanol, are added before granulation and drying. Then, ginger juice and distillate are added and mixed evenly, and then filler is added before tableting.

[0061] Then, HPLC was used to detect the reference solution and the test solution, and the chromatograms obtained were compared to determine the common characteristic peaks of Juanxiao tablets, and then the HPLC standard fingerprint chromatogram of Juanxiao tablets was determined.

[0062] The HPLC conditions include: a C18 column; a detection wavelength of 210-330 nm; for example, any value within the 210-330 nm range, such as 210 nm, 254 nm, 280 nm, 300 nm, and 330 nm; a column temperature of 28-32℃; for example, any value within the 28-32℃ range, such as 28℃, 29℃, 30℃, 31℃, and 32℃; and an injection volume of 5-10 μL; for example, any value within the 5-10 μL range, such as 5 μL, 6 μL, 7 μL, 8 μL, 9 μL, and 10 μL.

[0063] HPLC elution using a gradient of mobile phase content and flow rate is as follows: Mobile phase A is a nitrile solvent, preferably a C2-C5 nitrile solvent, more preferably a C2-C3 nitrile solvent, and most preferably acetonitrile. Mobile phase B is a 0.05-0.15% phosphoric acid solution, for example, any value between 0.05% and 0.15% phosphoric acid solution, such as 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.10%, 0.11%, 0.12%, 0.13%, 0.14%, and 0.15%.

[0064] The gradient elution process is as follows:

[0065]

[0066] Secondly, embodiments of the present invention provide a fingerprint spectrum for an asthma-relieving tablet, which is obtained by the above-described method for establishing an asthma-relieving tablet fingerprint spectrum.

[0067] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0068] The anti-asthma tablets provided by this invention are prepared using the following formula and process:

[0069] 418g of Lepidium apetalum, 418g of Citrus reticulata peel, 418g of Citrus reticulata peel, 625g of Vitex negundo fruit, 418g of Areca catechu, 125g of Rheum palmatum, and 100g of fresh ginger.

[0070] The above seven ingredients are prepared as follows: ginger juice is extracted, rhubarb is ground into a fine powder; dried tangerine peel, green tangerine peel, vitex fruit, areca nut, and lepidium seed (wrapped in cotton cloth) are steam distilled, and the distillate is collected; the distilled aqueous solution is filtered and set aside; the residue is boiled again with water, filtered, combined with the above filtrate, concentrated, cooled, and ethanol is added to make the alcohol content 65%, let stand for 24 hours, filtered, the ethanol is recovered from the filtrate, concentrated to obtain an extract, rhubarb fine powder is added, dried, ground into a fine powder, starch and ethanol are added as appropriate, granules are made, dried, ginger juice and distillate are sprayed in, mixed well, starch is added as appropriate, pressed into 1000 tablets, coated with a thin film, and the product is obtained.

[0071] Example 1

[0072] This invention provides a method for establishing a fingerprint spectrum of asthma tablets, as detailed below:

[0073] Instruments: Thermo Ultimate 3000 high performance liquid chromatograph; MSA224S-CE electronic balance; IQ7000 ultrapure water system.

[0074] The anti-asthma tablets were prepared in-house according to the above method, and the sample batch numbers are shown in Table 1. Acetonitrile and phosphoric acid were of chromatographic grade, water was ultrapure water, and all other reagents were of analytical grade.

[0075] Table 1. Sample Information of Anti-asthmatic Tablets

[0076]

[0077] Chromatographic conditions: Column: Agilent ZORBAX SB C18 column, 4.6 x 250 mm, particle size 5 μm; Mobile phase: Acetonitrile as mobile phase A, 0.10% phosphoric acid solution as mobile phase B, elution using flow rate gradient (see Table 2); Detection wavelength: 210-330 nm (283 nm); Column temperature: 30 °C; Injection volume: 5 μL.

[0078] Table 2 Gradient Elution

[0079]

[0080] Preparation of reference solutions: Weigh appropriate amounts of isoharmonin, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, nobiletin, and citrus tinctoria reference standards, dissolve them in methanol, and use them as reference solutions.

[0081] Preparation of test solution: Accurately weigh 0.5g of anti-asthmatic tablets, place them in a stoppered conical flask, add 75% methanol, sonicate for 30min, allow to cool naturally to room temperature, add 75% methanol to bring the weight back to the initial weight, filter, and collect the filtrate to obtain the test solution.

[0082] The test solution and reference solution were analyzed using the above chromatographic conditions. The HPLC fingerprints of 10 batches of Juanxiao tablets are shown below. Figure 1 Among them, S1-S10 are batch numbers: YF220705, YF230120, YF230712, YF230728, YF230809, YF230821, YF230829, YF231009, YF231026, and YF231119, respectively. The fingerprint similarity evaluation software "Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012.130723)" compiled by the Pharmacopoeia Commission was used for analysis and comparison. A common pattern chromatogram of the HPLC fingerprint of Juanxiao tablets was generated as the reference fingerprint chromatogram for Juanxiao tablets. (See...) Figure 2 , Figure 2 The following compounds were identified: 1. Isorhizin; 2. Isorhizin; 3. Rutin; 4. Hesperidin (S); 5. Lemon balm glycoside; 6. Aloe-emodin; 7. Norihesperidin; 9. Citrus reticulatae. Similarity results are shown in Table 3, and the relative retention times of shared patterns are shown in Table 4. The relative retention times of fingerprints from 10 batches of anti-asthmatic tablets are shown in Table 5.

[0083] according to Figure 1 and Figure 2 It can be seen that, from Figure 1 and Figure 2 It is known that the fingerprint spectrum of Juanxiao tablets has a total of 9 peaks, with peak 4 (hesperidin) serving as the reference peak (S peak). The relative retention times of each characteristic peak and peak S peak were calculated and are shown in Tables 3, 4, and 5. The relative retention times should be within ±10% of the specified values. The specified values ​​for the 8 common peaks in this spectrum are: 0.499, 0.634, 0.813, 1.641, 2.291, 2.427, 2.507, and 2.577.

[0084] Table 3. Similarity results of fingerprint patterns of anti-asthma tablets

[0085]

[0086] Table 4. Relative Retention Time of Common Patterns

[0087]

[0088]

[0089] Table 5. Relative retention times of fingerprint chromatograms from 10 batches of anti-asthma tablets.

[0090]

[0091] Based on the above results, using peak 4 (hesperidin) as the reference peak S in the fingerprint spectrum, the relative retention times of the common peaks 1-9 of Juanxiao tablets can be determined. This comprehensive and rapid testing of Juanxiao tablet samples in a single step is beneficial for the overall quality control of Juanxiao tablets, thereby contributing to improving the stability and safety of the drug.

[0092] Example 2-3

[0093] Examples 2-3 refer to the method provided in Example 1 to establish the fingerprint spectrum of the anti-asthmatic tablets. The only difference is the concentration of mobile phase B. Specifically, mobile phase B used in Example 2 is 0.05% phosphoric acid solution, and mobile phase B used in Example 3 is 0.15% phosphoric acid solution. The anti-asthmatic tablets used are anti-asthmatic tablets with batch number YF220705. All other operations are the same as in Example 1.

[0094] The detection results of Example 2 can be found here. Figure 3 The detection results of Example 3 Figure 4 ,according to Figure 3 and Figure 4 It can be seen that both 0.05% and 0.15% phosphoric acid solutions can achieve the detection of characteristic peaks, indicating that 0.05%-0.15% phosphoric acid solutions can be effectively used for the separation of components in Juanxiao tablets.

[0095] Examples 4-5

[0096] Examples 4-5 refer to the method provided in Example 1 to establish the fingerprint spectrum of the anti-asthmatic tablets. The only difference is the column temperature. The column temperature in Example 4 is 28°C and the column temperature in Example 5 is 32°C. The anti-asthmatic tablets used are anti-asthmatic tablets with batch number YF220705. All other operations are the same as in Example 1.

[0097] The detection results of Example 4 can be found here. Figure 5 The detection results of Example 5 Figure 6 ,according to Figure 5 and Figure 6 It can be seen that both column temperatures of 28℃ and 32℃ can achieve the detection of characteristic peaks, indicating that column temperatures of 28-32℃ can be effectively applied to the separation of components of Juanxiao tablets.

[0098] Verification Example

[0099] The precision, stability, and repeatability of the method provided in Example 1 were tested, as follows:

[0100] (1) Precision

[0101] Take the same sample solution (batch number: YF220705), and inject it 6 times under the chromatographic conditions of Example 1. The relative retention times of the common peaks are determined, and the results are shown in Table 6. The chromatogram for the precision test is shown in [Table 6]. Figure 7The similarity results are shown in Table 7.

[0102] Table 6. Results of Relative Retention Time in Precision Test

[0103]

[0104] Table 7. Similarity Results of Precision Test

[0105]

[0106] Based on the above results, the relative retention times (RSDs) of the common reference peak S peak (peak 4) were 0.10%, 0.07%, 0.06%, 0.06%, 0.07%, 0.06%, 0.05%, and 0.07%, respectively. The similarity to the fingerprint spectrum of the anti-asthmatic tablets was greater than 0.99, indicating good precision.

[0107] (2) Repeatability

[0108] Six test samples were prepared from the same test solution (batch number: YF220705). The relative retention times of the common peaks were determined under the chromatographic conditions described in Example 1. The results are shown in Table 8. The chromatograms for the repeatability test are shown in [Table 8]. Figure 8 The similarity results are shown in Table 9.

[0109] Table 8. Results of Relative Retention Time in Repeatability Tests

[0110]

[0111]

[0112] Table 9. Similarity Results of Repeatability Tests

[0113]

[0114] The results above show that the relative retention times (RSDs) of the common reference peak S peak (peak 4) are 0.04%, 0.03%, 0.04%, 0.02%, 0.03%, 0.03%, 0.03%, and 0.03%, respectively. The similarity to the fingerprint spectrum of the anti-asthmatic tablets is greater than 0.99, indicating that the method has good repeatability.

[0115] (3) Stability

[0116] Take the same sample solution (batch number: YF220705), and after preparation, inject it at 0, 2, 4, 6, 8, 10, 16, and 24 h according to the chromatographic conditions of Example 1. The relative retention times of the common peaks are determined, and the results are shown in Table 10. The chromatograms for the stability test are shown in [Table 10]. Figure 9 The similarity results are shown in Table 11.

[0117] Table 10 Results of relative retention times in stability tests

[0118]

[0119]

[0120] Table 11 Similarity Results of Stability Tests

[0121]

[0122] The results above show that the relative retention times (RSDs) of the common reference peak S (peak 4) were 0.09%, 0.06%, 0.05%, 0.06%, 0.07%, 0.06%, 0.06%, and 0.07%, respectively. The similarity to the fingerprint chromatogram of the antipyretic tablets was greater than 0.99, indicating that the test solution was stable within 24 hours and met the determination requirements.

[0123] Comparative Example 1

[0124] This comparative example provides a method for establishing the fingerprint spectrum of Juanxiao tablets. The operation is the same as in Example 1, except that the mobile phase used is different. Specifically, mobile phase A is methanol, mobile phase B is 0.1% phosphoric acid solution, and the Juanxiao tablets used are Juanxiao tablets with batch number YF220705. All other operations are the same as in Example 1.

[0125] See results Figure 10 ,Depend on Figure 10 It can be seen that the characteristic peaks were not sufficiently separated, therefore the mobile phase of methanol-0.1% phosphoric acid water is not suitable for the separation of the components of Juanxiao tablets. This shows that not any mobile phase is suitable for the separation and identification of the components of Juanxiao tablets.

[0126] Comparative Example 2

[0127] This comparative example provides a method for establishing the fingerprint spectrum of Juanxiao tablets. The operation is the same as in Example 1, except that the mobile phase used is different. Specifically, mobile phase A is acetonitrile, mobile phase B is 0.1% formic acid aqueous solution, and the Juanxiao tablets used are Juanxiao tablets with batch number YF220705. All other operations are the same as in Example 1.

[0128] See results Figure 11 ,according to Figure 11 It can be seen that the characteristic peaks were not sufficiently separated, and the mobile phase of acetonitrile-0.1% formic acid water is not suitable for the separation of the components of Juanxiao tablets. This shows that not any mobile phase is suitable for the separation and identification of the components of Juanxiao tablets.

[0129] Comparative Example 3

[0130] This comparative example provides a method for establishing the fingerprint spectrum of anti-asthmatic tablets. The operation is the same as that in Example 1, except that the gradient elution method used is different. For specific elution, please refer to Table 12. All other operations are the same as those in Example 1.

[0131] Table 12 Elution gradient

[0132]

[0133] See results Figure 12 ,according to Figure 12 It can be seen that the characteristic peaks were not sufficiently separated, and the elution gradient mobile phase is not suitable for the separation of the components of Juanxiao tablets. This indicates that not all gradient elution can achieve the separation and identification of the components of Juanxiao tablets, demonstrating that the gradient elution method provided in the embodiments of the present invention has beneficial effects.

[0134] Comparative Example 4

[0135] This comparative example provides a method for establishing the fingerprint spectrum of anti-asthmatic tablets. The operation is the same as that in Example 1, except that the gradient elution method used is different. For specific elution, please refer to Table 13. All other operations are the same as those in Example 1.

[0136] Table 13 Elution gradient

[0137]

[0138]

[0139] See results Figure 13 ,according to Figure 13 It can be seen that the characteristic peaks were not fully separated and the impurity peaks were obvious, indicating that this elution gradient mobile phase is not suitable for the separation of components in Juanxiao tablets.

[0140] Comparative Example 5

[0141] This comparative example provides a method for establishing the fingerprint spectrum of anti-asthmatic tablets. The operation is the same as that in Example 1, except that the gradient elution method used is different. For specific elution, please refer to Table 14. All other operations are the same as those in Example 1.

[0142] Table 14 Elution gradient

[0143]

[0144] See results Figure 14 ,according to Figure 14 It can be seen that the characteristic peaks were not fully separated and the impurity peaks were obvious, indicating that this elution gradient mobile phase is not suitable for the separation of components in Juanxiao tablets.

[0145] Comparative Example 6

[0146] This comparative example provides a method for establishing the fingerprint spectrum of anti-asthmatic tablets. The operation is the same as that in Example 1, except that the gradient elution method used is different. For specific elution, please refer to Table 15. All other operations are the same as those in Example 1.

[0147] Table 15 Elution gradient

[0148]

[0149] See results Figure 15 ,according to Figure 15 It can be seen that the characteristic peaks were not sufficiently separated, and this elution gradient mobile phase is not suitable for the separation of the components of the antihistamine tablets.

[0150] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for establishing a fingerprint spectrum of asthma tablets, characterized in that, include: Isocaroside, isovitexin, rutin, hesperidin, lemon balm glycoside, aloe-emodin, norihesperidin and citrus-reactive protein were used as reference standards, and anti-alzoster tablets were used as the test sample. The ingredients of the anti-asthmatic tablets are as follows, by weight: 418 parts of Lepidium apetalum, 418 parts of Citrus reticulata peel, 418 parts of Citrus reticulata peel, 625 parts of Vitex negundo fruit, 418 parts of Areca catechu, 125 parts of Rheum palmatum and 100 parts of Zingiber officinale. The reference standard and the test sample were detected by HPLC and then compared and analyzed to determine the common characteristic peaks of the anti-asthmatic tablets; The HPLC conditions included: a detection wavelength of 283 nm; a C18 column; mobile phase A of acetonitrile; mobile phase B of 0.05-0.15% phosphoric acid solution; and a gradient elution process as follows:

2. The method for establishing the fingerprint spectrum of asthma tablets according to claim 1, characterized in that, The mobile phase B is a 0.1% phosphoric acid solution.

3. The method for establishing the fingerprint spectrum of asthma tablets according to claim 1, characterized in that, HPLC conditions also include: column temperature of 28-32℃ and injection volume of 5-10μL.

4. The method for establishing the fingerprint spectrum of asthma tablets according to claim 1, characterized in that, Also includes: Before HPLC detection, the test sample and the reference sample are processed to form test sample solution and reference sample solution, respectively.

5. The method for establishing the fingerprint spectrum of asthma tablets according to claim 4, characterized in that, The steps for forming the test solution include: dissolving the anti-asthmatic tablets using an alcohol solvent, and performing ultrasonic treatment during the dissolution process.

6. The method for establishing the fingerprint spectrum of asthma tablets according to claim 5, characterized in that, include: For each gram of the aforementioned anti-asthmatic tablets, add 100-150 ml of alcohol solvent, mix and sonicate, cool after sonication and reweight to the initial weight, and then filter.

7. The method for establishing the fingerprint spectrum of asthma tablets according to claim 4, characterized in that, The steps for forming the reference solution include: dissolving the reference standard separately using an alcohol solvent.

8. The method for establishing the fingerprint spectrum of asthma tablets according to claim 1, characterized in that, The method for forming the aforementioned anti-asthma tablet is as follows: Ginger is juiced to make ginger juice; rhubarb is crushed to make rhubarb powder; The remaining raw materials are distilled using steam, and the distillate is collected. The distilled water is filtered to form the first filtrate. The residue after distillation is extracted with water and then filtered to form a second filtrate. The first filtrate and the second filtrate are mixed and then mixed with an alcohol solvent and allowed to stand. The mixture is then filtered to form a third filtrate, and the third filtrate is concentrated to form an extract powder. The extract powder is mixed with rhubarb powder and then pulverized. After adding fillers and humectants, it is granulated and dried. Then, ginger juice and distillate are added and mixed evenly, and then fillers are added and tableted.

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