Children qi depressing and asthma relieving mixture and quality detection method thereof

Abstract

The invention discloses a children qi depressing and asthma relieving mixture and a quality detection method thereof. The children qi depressing and asthma relieving mixture is prepared from honey-fried ephedra, scalded semen armeniacae amarae, fried fructus perillae, semen raphani, semen plantaginis, uncaria rhynchophylla, fried radix paeoniae alba and scutellaria baicalensis. According to the morbidity characteristics of the infantile asthma, the prepared infantile Qi-depressing asthma-relieving mixture can obviously relieve airway inflammation, improve lung functions and relieve airway hyperreaction, and has a good effect of treating the infantile asthma. According to the method, a detection method is screened through a large number of experiments, thin-layer chromatography qualitative identification and high performance liquid chromatography quantitative detection are adopted, ephedra sinica stapf, scalded semen armeniacae amarae, radix paeoniae alba and radix scutellariae are comprehensively and objectively detected and analyzed, and methodological investigation shows that the high performance liquid chromatography provided by the invention is high in precision and good in repeatability and stability. The traditional Chinese medicine composition has important significance for controlling the quality of the pediatric Qi depressing and calming asthma and ensuring the clinical curative effect.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and specifically relates to a Xiao'er Jiangqi Pingxiao Mixture for treating pediatric asthma and its quality detection method. Background Art

[0002] Bronchial asthma (hereinafter referred to as asthma) is the most common chronic airway disease in children. Repeated attacks of asthma can damage lung function and exacerbate airway injury, causing serious impacts on children's learning, daily activities, and even growth and development, and also bringing a heavy burden to families and society. Due to the complex western medicine treatment plan for asthma and poor compliance of children, the current daily management of asthma is not standardized, and the overall control level of pediatric asthma is still not ideal. Traditional Chinese medicine treatment of pediatric asthma has unique advantages in controlling the condition, reducing the severity of attacks, reducing the number of attacks, and improving the quality of life of children.

[0003] Therefore, the development of a traditional Chinese medicine preparation for treating the attack stage of pediatric asthma is of great significance for treating pediatric asthma. At present, the drug treatment of this disease generally uses chemical drugs and / or traditional Chinese medicine. Chemical drugs have a rapid onset of effect, but their treatment methods only target a certain pathogenesis of asthma. There are limitations in clinical treatment plan selection. In addition, the first-line inhalation therapy has high requirements for children's medication techniques, and long-term hormone maintenance treatment has certain side effects, resulting in poor medication compliance of children; traditional Chinese medicine treatment can treat both the symptoms and the root causes, is safe, effective, and has a lower treatment cost. The present invention is used to treat the external cold and internal heat syndrome in the attack stage of pediatric asthma, and is summarized based on years of clinical experience. It can well overcome the deficiency of the lack of similar marketed varieties of this syndrome type, and achieve the purpose of clinical control. Developing a clinically effective prescription into a safe and effective traditional Chinese medicine preparation can provide more choices for asthmatic children in terms of drug treatment.

[0004] Chinese Patent ZL201910400385.3 discloses a drug with the efficacy of treating pediatric asthma, its preparation method and application. The traditional Chinese medicine composition disclosed in this patent is made of honey-fried Ephedra, blanched Bitter Apricot Seed, stir-fried Perilla Seed, Radish Seed, Plantain Seed, Uncaria rhynchophylla, stir-fried White Peony Root, and Scutellaria baicalensis, and has a good therapeutic effect on pediatric asthma.

[0005] Xiao'er Jiangqi Pingxiao Mixture is an in-hospital traditional Chinese medicine preparation developed by Jiangsu Provincial Hospital of Traditional Chinese Medicine and has obtained the record of traditional Chinese medicine preparation. It is composed of eight traditional Chinese medicines: honey-fried Ephedra, blanched Bitter Apricot Seed, stir-fried Perilla Seed, Radish Seed, Plantain Seed, Uncaria rhynchophylla, stir-fried White Peony Root, and Scutellaria baicalensis. Clinical observation and research show that it has a good effect on treating pediatric asthma.

[0006] At present, there is no preparation and quality detection method for Xiao'er Jiangqi Pingxiao Mixture. The present invention screens out its thin-layer identification and high-performance liquid phase identification methods, which is of great significance for controlling its quality and ensuring clinical efficacy. Summary of the Invention

[0007] The object of the present invention is to provide a Xiao'er Jiangqi Pingxiao Mixture for treating pediatric asthma and its quality detection method.

[0008] To achieve the above object, the present invention provides the following technical solutions:

[0009] A Xiao'er Jiangqi Pingxiao Mixture is prepared by the following method:

[0010] (1) Take 1 - 8 parts by weight of honey - fried Ephedra, 6 - 14 parts by weight of blanched Bitter Apricot Seeds, 8 - 16 parts by weight of stir - fried White Peony Root, 6 - 14 parts by weight of Uncaria rhynchophylla, 6 - 14 parts by weight of Perilla Seed, 6 - 14 parts by weight of Plantain Seed, 6 - 14 parts by weight of Radish Seed, and 4 - 12 parts by weight of Scutellaria baicalensis Georgi, place them in an extraction tank, soak with water, decoct and extract multiple times, filter, combine the filtrates, take an appropriate amount of sucrose, dissolve it in water, mix it with the filtrate, and concentrate under reduced pressure;

[0011] (2) Centrifuge the concentrated liquid obtained in step (1) with a straight - tube high - speed centrifuge to obtain the mixture.

[0012] As a preferred scheme, the above - mentioned Xiao'er Jiangqi Pingxiao Mixture is prepared by the following method:

[0013] Take 1 - 8 parts by weight of honey - fried Ephedra, 6 - 14 parts by weight of blanched Bitter Apricot Seeds, 8 - 16 parts by weight of stir - fried White Peony Root, 6 - 14 parts by weight of Uncaria rhynchophylla, 6 - 14 parts by weight of Perilla Seed, 6 - 14 parts by weight of Plantain Seed, 6 - 14 parts by weight of Radish Seed, and 4 - 12 parts by weight of Scutellaria baicalensis Georgi, place them in an extraction tank, add 10 times the amount of water for the first time, soak for 0.5 h, decoct for 0.5 h, filter; add 8 times the amount of water for the second time, decoct for 0.5 h, filter; take an appropriate amount of sucrose, dissolve it in water, combine it with the two filtrates, concentrate under reduced pressure to a relative density of about 1.07 at 60 °C, place it in a straight - tube high - speed centrifuge, centrifuge at a rotation speed of 16000 r / min and a flow rate of 4 - 6 L / min to obtain the mixture.

[0014] As a more preferred scheme, the above - mentioned Xiao'er Jiangqi Pingxiao Mixture is made up of 4 parts by weight of honey - fried Ephedra, 10 parts by weight of blanched Bitter Apricot Seeds, 10 parts by weight of stir - fried Perilla Seed, 10 parts by weight of Radish Seed, 10 parts by weight of Plantain Seed, 10 parts by weight of Uncaria rhynchophylla, 12 parts by weight of stir - fried White Peony Root, and 8 parts by weight of Scutellaria baicalensis Georgi.

[0015] The quality detection method of the Xiao'er Jiangqi Pingxiao Mixture for treating pediatric asthma according to the present invention includes thin - layer chromatography qualitative detection and high - performance liquid chromatography quantitative detection.

[0016] Preferably, the thin - layer chromatography qualitative detection includes the following steps:

[0017] (1) Identification of Ephedra by thin - layer chromatography, the specific steps are as follows:

[0018] 1) Take Xiaoer Jiangqi Pingxiao mixture, add a few drops of concentrated ammonia, extract with dichloromethane, combine the dichloromethane solutions, evaporate to dryness in a water bath, dissolve the residue in methanol, and use it as the test solution. Take the traditional Chinese medicine compound composition lacking ephedra and prepare an ephedra-negative solution in the same way;

[0019] 2) Separately, take Ephedra control medicinal material powder, add a few drops of concentrated ammonia test solution, then add chloroform, heat under reflux, filter, evaporate the filtrate to dryness, and dissolve the residue in methanol to prepare the Ephedra control medicinal material solution;

[0020] 3) Pipette the three solutions above and spot them separately on a single silica gel G thin-layer plate. Develop with chloroform-methanol-concentrated ammonia. Remove, air-dry, spray with ninhydrin test solution, and heat until the spots are clearly colored. If the test sample chromatogram shows a spot of the same color at the corresponding position in the control chromatogram, no interference is observed in the negative assay.

[0021] (2) Thin layer chromatography identification of braised bitter almonds, the specific steps are as follows:

[0022] 1) Take Xiaoer Jiangqi Pingxiao mixture, add anhydrous ethanol, let stand, filter, evaporate the filtrate to dryness, dissolve in water, extract with water-saturated n-butanol in batches, combine the n-butanol layers, evaporate to dryness, and dissolve the residue in methanol to prepare the test solution. Take the traditional Chinese medicine compound composition without boiled bitter almonds and prepare a boiled bitter almond negative solution in the same way;

[0023] 2) Prepare another amygdalin reference solution by adding methanol to prepare a reference solution containing 2 mg of amygdalin per 1 ml.

[0024] 3) Pipette each of the three solutions onto a single silica gel GF254 thin layer plate. Develop with ethyl acetate-methanol-water. Remove, allow to air dry, and spray with anisaldehyde-sulfuric acid chromogen solution. Heat until the spots are clearly colored. If the test solution chromatogram shows a spot of the same color at the corresponding position in the control solution chromatogram, no interference is observed in the negative sample.

[0025] (3) Thin layer chromatography identification of white peony root, the specific steps are as follows:

[0026] 1) Take Xiaoer Jiangqi Pingxiao mixture, evaporate to dryness, add ethanol to ultrasonic treatment, filter, evaporate the filtrate to dryness, and dissolve the residue in ethanol to prepare the test solution; take the traditional Chinese medicine compound composition without white peony root, and prepare the white peony root negative solution in the same way;

[0027] 2) Separately, take a white peony root control medicinal material, add ethanol and shake, filter, evaporate the filtrate to dryness, and dissolve the residue in ethanol to prepare the white peony root control medicinal material solution;

[0028] 4) Take the paeoniflorin reference substance and add ethanol to make a solution containing 1 mg per 1 ml as the reference solution;

[0029] 3) Pipette each of the four solutions onto a single silica gel G thin layer plate. Develop with chloroform-ethyl acetate-methanol-formic acid. Remove, allow to air dry, spray with vanillin-sulfuric acid solution, and heat until the spots are clearly colored. If the test solution chromatogram shows spots of the same color at corresponding positions in the chromatograms of the control herb and reference solution solutions, no interference is observed with the negative results.

[0030] (4) Thin layer chromatography identification of Scutellaria baicalensis, the specific steps are as follows:

[0031] 1) Take Xiaoer Jiangqi Pingxiao mixture, adjust the pH to 2-3 with dilute hydrochloric acid, extract with ethyl acetate, combine the two ethyl acetate extracts, evaporate to dryness, and dissolve the residue in methanol to prepare the test solution; take the traditional Chinese medicine compound composition lacking Scutellaria baicalensis and prepare a Scutellaria baicalensis negative solution in the same manner;

[0032] 2) Separately, take a reference herb of Scutellaria baicalensis, add ethyl acetate-methanol, heat to reflux, cool, filter, evaporate the filtrate to dryness, dissolve the residue in methanol, and use the supernatant as the reference herb solution of Scutellaria baicalensis;

[0033] 3) Pipette the above three solutions and spot them on the same polyamide film respectively. Use toluene-ethyl acetate-methanol-formic acid as the developing agent, develop, take out, dry, spray with aluminum chloride solution, dry, and examine under ultraviolet light. In the chromatogram of the test solution, at the corresponding position in the chromatogram of the control medicinal material solution, spots of the same color appear under fluorescence, indicating no interference when negative.

[0034] As a preferred embodiment, the quality detection method of the above-mentioned pediatric Jiangqi Pingxiao mixture for treating childhood asthma, step (1) thin layer chromatography identification of ephedra, using chloroform-methanol-concentrated ammonia in a volume ratio of 20:5:0.5 as the developing solvent;

[0035] Step (2) identification by thin layer chromatography of bitter almonds using ethyl acetate-methanol-water in a volume ratio of 8:1:1 as the developing solvent;

[0036] Step (3) thin layer chromatography identification of white peony root, using chloroform-ethyl acetate-methanol-formic acid in a volume ratio of 40:5:10:0.2 as a developing solvent;

[0037] Step (4) identification of Scutellaria baicalensis by thin layer chromatography using toluene-ethyl acetate-methanol-formic acid in a volume ratio of 10:3:1:2 as a developing solvent.

[0038] As a preferred embodiment, the quality detection method of the above-mentioned pediatric Jiangqipingxiao mixture for treating childhood asthma, wherein the high performance liquid chromatography quantitative detection comprises the following steps:

[0039] (1) Preparation of reference substance solution: Weigh accurately appropriate amounts of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride reference substances, dissolve them with methanol, and prepare single reference substance solutions with mass concentrations of 1.01, 0.89, 1.45, 0.94, and 0.97 mg·ml -1 respectively. Accurately pipette appropriate amounts of the above reference substance solutions into a 20-ml volumetric flask, dilute to the mark with methanol, and prepare a mixed reference substance solution with mass concentrations of 0.051, 0.134, 0.058, 0.047, and 0.048 mg·ml -1 respectively;

[0040] (2) Preparation of test solution: Accurately pipette 2 ml of the Xiao'er Jiangqi Pingxiao Mixture sample, dilute to the mark with 70% methanol in a 10-ml volumetric flask. Perform ultrasonic extraction (power 100 W, frequency 40 KHz) for 30 min, weigh, make up the weight with 70% methanol, shake well, centrifuge for 10 min (3000 r / min), take the supernatant, filter, and obtain the test solution;

[0041] (3) Plotting of standard curve: Take the reference substance solution in step (1) above, dilute it step by step with methanol, accurately pipette and inject it separately, and inject it into a high-performance liquid chromatograph for determination; take the mass concentration X of the reference substance as the abscissa and the peak area Y as the ordinate to obtain the regression equations of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride;

[0042] (4) Content determination: Accurately pipette the reference substance solution in step (1) and the test solution in step (2) respectively, inject them into the liquid chromatograph, and calculate the contents of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride in the test solution according to the standard curve equation in step (3).

[0043] As a preferred solution, for the quality detection method of the Xiao'er Jiangqi Pingxiao Mixture for treating pediatric asthma described above, the chromatographic conditions in step (4) are as follows: chromatographic column Agilent SB-Phenyl (4.6 mm × 250 mm, 5 μm); acetonitrile (A)-0.2% phosphoric acid aqueous solution (B) as the mobile phase; gradient elution (0 - 11 min, 5% → 5% A; 11 - 12 min, 5% → 8% A; 12 - 15 min, 8% → 8% A; 15 - 26 min, 8% → 20% A; 26 - 30 min, 20% → 23% A; 30 - 38 min, 23% → 40% A); column temperature 25°C; detection wavelength 210 nm; flow rate 1 ml·min -1 ; injection volume 10 μL.

[0044] Compared with the prior art, the beneficial effects of the present invention are:

[0045] 1. The present invention is formulated according to the pathogenesis characteristics of pediatric asthma and the principles of syndrome differentiation and treatment in traditional Chinese medicine. It has the effects of lowering qi and relieving asthma, relieving cough and resolving phlegm, and relieving exterior syndrome and clearing interior heat. It is mainly used for children with mild to moderate external cold and internal heat syndrome during asthma attacks. Symptoms include paroxysmal cough, expectoration of thick and sticky phlegm that is yellow or white in color, shortness of breath, accompanied by nasal congestion, sneezing, runny nose, or sometimes aversion to cold, fever without sweating, restlessness at night, and dry stools. In the formula, honey-fried ephedra is the first drug for treating asthma, which is good at relieving exterior cold and expelling wind and relieving asthma; stir-fried bitter apricot seeds lower and promote the flow of lung qi. The two are used as the monarch drugs together, with the promotion and descent working in coordination to make the lung qi smooth. Perilla seed, radish seed, and plantain seed are used as the ministerial drugs together. The perilla seed of the new "Three Seeds" expels phlegm in the upper jiao and also descends lung qi, the radish seed eliminates phlegm in the middle jiao and also awakens the spleen earth, and the plantain seed clears lung heat and resolves phlegm, diffusing and guiding pathogenic factors outwards. The adjuvant drugs include scutellaria baicalensis to clear lung fire, dry dampness, and resolve phlegm, white peony root to astringe yin and harmonize nutrient qi to assist the defensive qi, and uncaria rhynchophylla to gently clear and expel pathogenic factors to dispel latent wind. The whole formula coordinates ascending and descending, with descending as the main, combines clearing and warming, and treats phlegm and qi simultaneously, jointly achieving the effects of lowering qi and relieving asthma, relieving cough and resolving phlegm, and relieving exterior syndrome and clearing interior heat. Through a large number of experiments, the best raw material composition and dosage ratio are screened out, and the traditional Chinese medicine compound obtained by formulation can significantly improve the symptoms of asthmatic children and has a good effect on treating pediatric asthma.

[0046] 2. The present invention screens and detects methods through a large number of experiments. By using thin-layer chromatography for qualitative identification and high-performance liquid chromatography for quantitative detection, it can comprehensively and objectively detect and analyze ephedra, bitter apricot seeds, white peony root, and scutellaria baicalensis in the formula. Moreover, methodological detection shows that the high-performance liquid chromatography method provided by the present invention has high precision, good repeatability, and stability. Detailed implementation mode

[0047] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts fall within the protection scope of the present invention. Description of the drawings

[0048] Figure 1 Schematic diagram of thin-layer identification of different sample application amounts of ephedra in the present invention;

[0049] Figure 2 Schematic diagram of thin-layer identification of ephedra in Xiao'er Jiangqi Pingxiao Mixture under n-butanol - glacial acetic acid - water (8:2:1) in the present invention;

[0050] Figure 3 Schematic diagram of thin-layer identification of ephedra in Xiao'er Jiangqi Pingxiao Mixture under chloroform - methanol - concentrated ammonia (20:5:0.5) in the present invention;

[0051] Figure 4 Schematic diagram of thin-layer identification of different sample application amounts of bitter apricot seeds in the present invention;

[0052] Figure 5 This is a schematic diagram of the thin-layer identification of bitter apricot kernels in the Xiao'er Jiangqi Pingxiao Mixture under dichloromethane - methanol - glacial acetic acid - water (20:10:3:1) according to the present invention;

[0053] Figure 6 This is a schematic diagram of the thin-layer identification of bitter apricot kernels in the Xiao'er Jiangqi Pingxiao Mixture under ethyl acetate - methanol - water (8:1:1) according to the present invention;

[0054] Figure 7 This is a schematic diagram of the thin-layer identification of Paeonia lactiflora Pall. with different sample application amounts according to the present invention;

[0055] Figure 8 This is a schematic diagram of the thin-layer identification of Paeonia lactiflora Pall. in the Xiao'er Jiangqi Pingxiao Mixture under chloroform - ethyl acetate - methanol - formic acid (40:5:10:0.2) according to the present invention;

[0056] Figure 9 This is a schematic diagram of the thin-layer identification of Paeonia lactiflora Pall. in the Xiao'er Jiangqi Pingxiao Mixture under chloroform - ethyl acetate - methanol - concentrated ammonia (8:1:4:1) according to the present invention;

[0057] Figure 10 This is a schematic diagram of the thin-layer identification of Scutellaria baicalensis Georgi with different sample application amounts according to the present invention;

[0058] Figure 11 This is a schematic diagram of the thin-layer identification of Scutellaria baicalensis Georgi in the Xiao'er Jiangqi Pingxiao Mixture under cyclohexane - ethyl acetate - formic acid (7:3:0.2) according to the present invention;

[0059] Figure 12 This is a schematic diagram of the thin-layer identification of Scutellaria baicalensis Georgi in the Xiao'er Jiangqi Pingxiao Mixture under toluene - ethyl acetate - methanol - formic acid (10:3:1:2) according to the present invention;

[0060] Figure 13 This is a schematic diagram of the specificity chromatogram of the test solution of the Xiao'er Jiangqi Pingxiao Mixture according to the present invention. In the figure, a. test solution, b. Scutellaria baicalensis Georgi negative, c. honey-fried Ephedra sinica Stapf negative, d. stir-fried Paeonia lactiflora Pall. negative, e. blanched bitter apricot kernels negative, f. mixed reference substance solution; 1. ephedrine hydrochloride, 2. pseudoephedrine hydrochloride, 3. amygdalin, 4. paeoniflorin, 5. baicalin.

[0061] Preparation method of the Xiao'er Jiangqi Pingxiao Mixture in Example 1, specifically including the following steps:

[0062] Take 4g of honey-fried Ephedrae Herba, 10g of blanched Armeniacae Semen Amarum, 10g of stir-fried Perillae Semen, 10g of Raphanus Sativae Semen, 10g of Plantaginis Semen, 10g of Uncariae Ramulus cum Uncis, 12g of stir-fried Paeoniae Radix Alba, and 8g of Scutellariae Radix. Place them in an extraction tank, decoct with water twice. For the first time, add 10 times the amount of water, soak for 0.5h, decoct for 0.5h, and filter; for the second time, add 8 times the amount of water, decoct for 0.5h, and filter. Take 4g of sucrose, dissolve it in water, combine it with the two filtrates, and concentrate under reduced pressure to a relative density of about 1.07 (60°C). Centrifuge with a straight-tube high-speed centrifuge (rotation speed: 16,000 r / min, flow rate: 4 - 6 L / min) to obtain the mixture.

[0063] Example 2 Thin-layer chromatography identification of the Pediatric Qi-Dropping and Wheezing-Relieving Mixture. The Pediatric Qi-Dropping and Wheezing-Relieving Mixture is prepared according to Example 1. The specific steps are as follows:

[0064] Step 1: Thin-layer chromatography identification of Ephedrae Herba. The specific steps are as follows:

[0065] (1) Take 12.5 ml of the Pediatric Qi-Dropping and Wheezing-Relieving Mixture, add a few drops of concentrated ammonia, shake and extract with dichloromethane 2 times, 30 ml each time. Combine the two dichloromethane solutions, evaporate to dryness in a water bath, dissolve the residue in 1 ml of methanol to obtain the test solution. Take the other herbs without Ephedrae Herba in the traditional Chinese medicine composition according to Example 1 and prepare the negative solution without Ephedrae Herba in the same way.

[0066] (2) Take another 1g of Ephedrae Herba control medicinal material, add a few drops of concentrated ammonia test solution, then add 10 ml of chloroform, heat under reflux for 1 hour, filter, evaporate the filtrate to dryness, dissolve the residue in 2 ml of methanol, filter, and take the filtrate as the Ephedrae Herba control medicinal material solution.

[0067] (3) Absorb the above three solutions and spot them on the same silica gel G thin-layer plate respectively. Use chloroform - methanol - concentrated ammonia (20:5:0.5) as the developing agent, develop, take out, dry in the air, spray with 2% ninhydrin test solution, and heat until the spots are clearly visible. In the test solution chromatogram, at the position corresponding to the control chromatogram, there are spots of the same color, and there is no interference in the negative.

[0068] (4) Durability detection of the Ephedrae Herba thin-layer method:

[0069] Different sample application amounts: Detect the feasibility of the thin-layer identification method of Ephedrae Herba in the Pediatric Qi-Dropping and Wheezing-Relieving Mixture under the conditions of 5 μL of Ephedrae Herba negative, 5 μL of ephedrine hydrochloride, and 5 μL and 4 μL of the test sample application. The results show that when the sample application amount is 5 μL, the identification spots are clearer; see Figure 1 , where Figure 1 1 is the Ephedrae Herba negative solution, 2 is the control medicinal material, and 3 and 4 are the test samples.

[0070] Comparison of the developing effects of different developing agents: The thin-layer method of Ephedrae Herba uses n-butanol - glacial acetic acid - water (8:2:1) and chloroform - methanol - concentrated ammonia (20:5:0.5) as the developing agents. The results are shown inFigure 2 , 3 . As can be seen from the figure, the development effect of n-butanol - glacial acetic acid - water (8:2:1) is poor, so chloroform - methanol - concentrated ammonia (20:5:0.5) is still used as the developing agent. Figure 2 , 3 In 3 , 1 is the negative solution of ephedra, 2 is the control medicinal material, and 3 and 4 are the test samples.

[0071] Different temperatures: The development conditions of the thin-layer identification of ephedra were detected at 8°C and 25°C respectively by using the above method. The results showed no obvious difference between the two temperatures, so this method is applicable to the thin-layer identification of ephedra in Xiao'er Jiangqi Pingxiao Mixture at different temperatures.

[0072] Different humidities: The feasibility of the thin-layer identification method of ephedra in Xiao'er Jiangqi Pingxiao Mixture was detected at humidities of 35% and 56% respectively. The results showed no obvious difference between the two humidities, and there were obvious and clear thin-layer identification points, so this method is applicable to the thin-layer identification of ephedra at different humidities.

[0073] Different manufacturers' thin-layer plates: Kangyexin thin silica gel G plates and Qingdao Ocean silica gel G plates were used for development respectively. The results showed no obvious difference, so this method has good adaptability and has no requirements for the manufacturers and brands of thin-layer plates.

[0074] Different batches of thin-layer plates: Silica gel G plates of the same manufacturer with different batches (20190506, 20201226) were used for development respectively. The results showed no obvious difference, so this method has good adaptability and has no requirements for the batches of thin-layer plates.

[0075] Step 2: The thin-layer chromatography identification of bitter apricot kernels is carried out as follows:

[0076] (1) Take 10 ml of Xiao'er Jiangqi Pingxiao Mixture, add 15 ml of absolute ethanol, let it stand for 12 h, filter, evaporate the filtrate to dryness, dissolve it in 10 ml of water, extract it with 6 times of water-saturated n-butanol in portions, combine the n-butanol layers, evaporate the n-butanol to dryness, add 2 ml of methanol to dissolve it, and use it as the test sample solution. Take other traditional Chinese medicine raw materials without bitter apricot kernels according to the traditional Chinese medicine composition in Example 1, and prepare the negative solution of bitter apricot kernels in the same way;

[0077] (2) Take another 20 mg of amygdalin reference substance, add 10 ml of methanol, and ultrasonically treat it for 10 minutes to obtain the amygdalin reference substance solution.

[0078] (3) Absorb the above 3 kinds of solutions, and spot them on the same silica gel GF254 thin-layer plate respectively. Use ethyl acetate - methanol - water (8:1:1) as the developing agent, develop, take out, dry in the air, spray with anisaldehyde sulfuric acid developer, and heat at 105°C until the spots are clearly developed, and no interference is seen in the negative.

[0079] (4) Durability detection of the thin-layer method for bitter apricot kernels

[0080] Different sample application volumes: The feasibility of the thin-layer identification method for Armeniacae Semen Amarum in Xiao'er Jiangqi Pingxiao Mixture was detected under the conditions of spotting 5 μL of amygdalin reference substance and 3 μL, 5 μL, and 8 μL of the test sample. As a result, when spotting 8 μL, the identification spots were clearer. See Figure 4 , where Figure 4 1 in

[0081] is the amygdalin reference substance, and 2, 3, and 4 are the test samples. Figure 5 、 6 . Finally, ethyl acetate - methanol - water (8:1:1) with clear spots was selected as the developing agent. In Figure 5 、 Figure 6 , 1 is the negative solution of Armeniacae Semen Amarum, 2 is the amygdalin reference substance, and 3 and 4 are the test samples.

[0082] Different temperatures: The developing conditions for the thin-layer identification of Armeniacae Semen Amarum were detected at 8 °C and 25 °C respectively using the above method. As a result, there was no significant difference between the two temperatures. Therefore, this method is applicable to the thin-layer identification of Armeniacae Semen Amarum in Xiao'er Jiangqi Pingxiao Mixture at different temperatures.

[0083] Different humidities: The feasibility of the thin-layer identification method for Armeniacae Semen Amarum in Xiao'er Jiangqi Pingxiao Mixture was detected at humidities of 35% and 56% respectively. As a result, there was no significant difference between the two humidities, and there were obvious and clear thin-layer identification spots. Therefore, this method is applicable to the thin-layer identification of Armeniacae Semen Amarum at different humidities.

[0084] Different manufacturers' thin-layer plates: Development was carried out using Kangyexin silica gel GF254 plates and Qingdao Ocean silica gel GF254 plates respectively. As a result, there was no significant difference. Therefore, this method has good adaptability and has no requirements for the manufacturer and brand of the thin-layer plate.

[0085] Different batch numbers of thin-layer plates: Development was carried out using silica gel G plates of the same manufacturer with different batch numbers (20190708, 20210418) respectively. As a result, there was no significant difference. Therefore, this method has good adaptability and has no requirements for the batch number of the thin-layer plate.

[0086] Step 3: For the thin-layer chromatography identification of Paeoniae Radix Alba, the specific steps are as follows:

[0087] (1) Take 5 ml of Xiao'er Jiangqi Pingxiao Mixture, evaporate to dryness, add 10 ml of ethanol, extract ultrasonically for 30 minutes, filter, evaporate the filtrate to dryness, and dissolve the residue in 1 ml of ethanol to obtain the test sample solution. According to the Chinese medicinal composition in Example 1, take the other Chinese medicinal raw materials without Paeoniae Radix Alba and prepare the negative test sample solution of Paeoniae Radix Alba in the same way.

[0088] (2) Separately take 0.5 g of the control crude drug of Paeonia lactiflora Pall., add 10 ml of ethanol, shake for 5 minutes, filter, evaporate the filtrate to dryness, dissolve the residue in 1 ml of ethanol to obtain the control crude drug solution.

[0089] (3) Take paeoniflorin reference substance, add ethanol to make a solution containing 1 mg per 1 ml as the reference substance solution.

[0090] (4) Absorb the above 4 kinds of solutions, respectively spot them on the same silica gel G thin-layer plate, use chloroform - ethyl acetate - methanol - formic acid (40:5:10:0.2) as the developing agent, develop, take out, dry in air, spray with 5% vanillin sulfuric acid solution. In the chromatogram of the test solution, at the corresponding position of the chromatogram of the control crude drug solution, under fluorescence, spots of the same color appear, and there is no interference in the negative sample.

[0091] (5) Durability test of the thin-layer method for Paeonia lactiflora Pall.

[0092] Different sample application amounts: Detect the feasibility of the thin-layer identification method of Paeonia lactiflora Pall. in Xiao'er Jiangqi Pingxiao Mixture under the conditions of 10 μL of Paeonia lactiflora Pall. negative sample, 10 μL of paeoniflorin, 10 μL and 8 μL of the test sample spotting respectively. As a result, when spotting 10 μL, the identification spots are clearer. See Figure 7 , among which Figure 7 1 is the Paeonia lactiflora Pall. negative sample, 2 is paeoniflorin, 3 and 4 are the test samples.

[0093] Comparison of the developing effects of different developing agents: The thin-layer method of Paeonia lactiflora Pall. uses chloroform - ethyl acetate - methanol - formic acid (40:5:10:0.2) and chloroform - ethyl acetate - methanol - concentrated ammonia (8:1:4:1) as the developing agents respectively. The results are shown in Figure 8 , 9 . Finally, chloroform - ethyl acetate - methanol - formic acid (40:5:10:0.2) with clear spots is selected as the developing agent. Among which Figure 8 , 9 1 is the Paeonia lactiflora Pall. negative solution, 2 is the control crude drug of Paeonia lactiflora Pall., 3 is the test sample, and 4 is the paeoniflorin reference substance.

[0094] Different temperatures: Detect the developing conditions of the thin-layer identification of Paeonia lactiflora Pall. by the above method at 8 °C and 25 °C respectively. As a result, there is no obvious difference between the two temperatures, so this method is applicable to the thin-layer identification of Paeonia lactiflora Pall. in Xiao'er Jiangqi Pingxiao Mixture at different temperatures.

[0095] Different humidities: Detect the feasibility of the thin-layer identification method of Paeonia lactiflora Pall. in Xiao'er Jiangqi Pingxiao Mixture at humidities of 35% and 56% respectively. As a result, there is no obvious difference between the two humidities, and there are obvious and clear thin-layer identification spots, so this method is applicable to the thin-layer identification of Paeonia lactiflora Pall. at different humidities.

[0096] Thin layer plates from different manufacturers: The plates from Kangyexin silica gel G and Qingdao Ocean silica gel G were used for development respectively, and there were no obvious differences in the results. Therefore, this method has good adaptability and has no requirements for the manufacturers and brands of thin layer plates.

[0097] Thin layer plates with different batch numbers: Silica gel G plates with different batch numbers (20190506, 20201226) from the same manufacturer were used for development respectively, and there were no obvious differences in the results. Therefore, this method has good adaptability and has no requirements for the batch numbers of thin layer plates.

[0098] Step 4: The thin layer chromatography identification of Scutellaria baicalensis is as follows:

[0099] (1) Take 10 ml of Xiao'er Jiangqi Pingxiao Mixture, adjust the pH to 2 with 6 ml / mol dilute hydrochloric acid, extract twice with 20 ml of ethyl acetate by shaking, combine the two ethyl acetate layers (upper layer), evaporate to dryness, dissolve the residue in 2 ml of methanol to obtain the test solution. According to the traditional Chinese medicine composition in Example 1, other traditional Chinese medicine raw materials without Scutellaria baicalensis were taken, and the Scutellaria baicalensis negative solution was prepared in the same way.

[0100] (2) Separately take 1 g of Scutellaria baicalensis control medicinal material, add 30 ml of ethyl acetate - methanol (3:1), heat under reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, dissolve the residue in 5 ml of methanol, and take the supernatant to obtain the control medicinal material solution.

[0101] (3) Pipette the above three solutions respectively and spot them on the same polyamide thin layer. Use cyclohexane - dichloromethane - ethyl acetate (10:5:8) as the developing agent, develop, take out, dry in air, and examine under an ultraviolet lamp (365 nm). In the chromatogram of the test solution, at the corresponding position of the chromatogram of the control medicinal material solution, spots of the same color appear under fluorescence, and there is no interference in the negative solution.

[0102] (4) Durability detection of the thin layer method for Scutellaria baicalensis

[0103] Different sample application volumes: The feasibility of the thin layer identification method for Scutellaria baicalensis in Xiao'er Jiangqi Pingxiao Mixture was detected respectively under the conditions of 5 μL of Scutellaria baicalensis negative, 3 μL of control medicinal material, and 3 μL, 2 μL, and 1 μL of the test sample application. The results are shown in Figure 10 , when the sample application volume is 3 μL, the identification spots are clearer, where Figure 10 1 is the Scutellaria baicalensis negative solution, 2 is the control medicinal material, and 3, 4, and 5 are test samples of different batches.

[0104] Comparison of the development effects of different developing agents: The thin layer method for Scutellaria baicalensis uses cyclohexane - ethyl acetate - formic acid (7:3:0.2) and toluene - ethyl acetate - methanol - formic acid (10:3:1:2) as the developing agents respectively, and the results are shown in Figure 11 , 12 . Finally, toluene - ethyl acetate - methanol - formic acid (10:3:1:2) with clear corresponding bands and more numbers was selected as the developing agent. Among themFigure 11 , 12 1 is the Scutellaria baicalensis negative solution, 2 is the Scutellaria baicalensis control medicinal material, and 3 - 5 are the test samples.

[0105] Different temperatures: The development conditions of the thin-layer identification of Scutellaria baicalensis were detected at 8°C and 25°C respectively using the above method. The results showed no significant difference between the two temperatures, so this method is applicable to the thin-layer identification of Scutellaria baicalensis in Xiao'er Jiangqi Pingxiao Mixture at different temperatures.

[0106] Different humidities: The feasibility of the thin-layer identification method of Scutellaria baicalensis in Xiao'er Jiangqi Pingxiao Mixture was detected at humidities of 35% and 56% respectively. The results showed no significant difference between the two humidities, and there were obvious and clear thin-layer identification points, so this method is applicable to the thin-layer identification of Scutellaria baicalensis at different humidities.

[0107] Different manufacturers' thin-layer plates: The development was carried out using the polyamide film from Luqiao and the polyamide film from Guoyao respectively. The results showed no significant difference, so this method has good adaptability and has no requirements for the manufacturer and brand of the thin-layer plate.

[0108] Different batch numbers of thin-layer plates: The development was carried out using polyamide films with different batch numbers (20190905, 20210226) from the same manufacturer respectively. The results showed no significant difference, so this method has good adaptability and has no requirements for the batch number of the thin-layer plate.

[0109] Example 3 Content determination method of Xiao'er Jiangqi Pingxiao Mixture

[0110] (1) Chromatographic conditions: Chromatographic column: Agilent SB-Phenyl (4.6 mm × 250 mm, 5 μm); Mobile phase: Acetonitrile (A) - 0.2% phosphoric acid aqueous solution (B); Gradient elution (0 - 11 min, 5% → 5% A; 11 - 12 min, 5% → 8%

[0111] A; 12 - 15 min, 8% → 8% A; 15 - 26 min, 8% → 20% A; 26 - 30 min, 20% → 23% A; 30 - 38 min, 23% → 40% A); Column temperature: 25°C; Detection wavelength: 210 nm; Flow rate: 1 ml·min -1 ; Injection volume: 10 μL.

[0112] (2) Preparation of reference solution: Weigh accurately appropriate amounts of reference substances of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride, dissolve them with methanol, and prepare single reference solutions with mass concentrations of 1.01, 0.89, 1.45, 0.94, and 0.97 mg·ml -1 respectively. Precisely pipette appropriate amounts of the above reference solutions into a 20 ml volumetric flask, dilute to the mark with methanol, and prepare a reference solution containing mass concentrations of 0.051, 0.134, 0.058, 0.047, and 0.048 mg·ml-1 Mixed reference substance solution.

[0113] (3) Preparation of test solution: Precisely measure 2 ml of the Xiao'er Jiangqi Pingxiao Mixture prepared in Example 1, add 70% methanol to make up the volume to a 10-ml volumetric flask. Ultrasonically extract (power 100 W, frequency 40 KHz) for 30 min, weigh, make up the weight with 70% methanol, shake well, centrifuge for 10 min (3000 r / min), take the supernatant, filter, and obtain. And prepare negative test solutions lacking honey-fried ephedra, blanched bitter apricot seeds, stir-fried white peony root, stir-fried perilla seeds, scutellaria baicalensis, uncaria rhynchophylla, stir-fried radish seeds, and semen plantaginis praeparatum under the same conditions as in Example 1.

[0114] (4) Investigation of linear relationship:

[0115] Take appropriate amounts of each reference substance, dissolve in methanol, and dilute step by step. Determine according to the above chromatographic conditions. Set the abscissa (X) as the mass concentration of the reference substance and the ordinate (Y) as the peak area, and perform linear regression analysis. The results are shown in Table 1.

[0116] Table 1 Results of investigation of linear relationship

[0117] Index component Regression equation <![CDATA[R 2 > <![CDATA[Linear range (μg·ml -1 )]]> Ephedrine Y = 16.17X + 8.5526 0.9999 6.072～219.120 Pseudoephedrine Y = 16.691X + 8.941 0.9999 5.780～172.092 Amygdalin Y = 11.945X + 3.6842 0.9994 16.612～526.020 Paeoniflorin Y = 3.6332X + 0.9838 0.9996 18.738～674.556 Baicalin Y = 33.302X + 38.182 0.9994 41.293～1486.548

[0118] (5) Precision test:

[0119] Take 10 μL of the same reference substance solution and inject it into the high-performance liquid chromatograph, and repeat the determination 6 times. Calculate with the peak area integral value. The RSD values of the peak areas of ephedrine hydrochloride, pseudoephedrine hydrochloride, amygdalin, paeoniflorin, and baicalin are 1.50%, 1.04%, 1.50%, 1.67%, and 1.13% respectively, indicating good instrument precision.

[0120] (6) Repeatability test:

[0121] Precisely weigh the mixture prepared according to the method in Example 1, a total of 6 portions, prepare the sample solution according to the above requirements, and determine according to law. The RSDs of the peak areas of ephedrine hydrochloride, pseudoephedrine hydrochloride, amygdalin, paeoniflorin, and baicalin are 1.91%, 1.44%, 0.74%, 1.03%, and 0.55% respectively, indicating good repeatability of this method.

[0122] (7) Stability test:

[0123] Take the same mixture and inject samples at 0, 2, 4, 8, 12, and 24 h after the preparation of the sample solution is completed, and determine. The RSDs of the peak areas of ephedrine hydrochloride, pseudoephedrine hydrochloride, amygdalin, paeoniflorin, and baicalin are 1.33%, 1.50%, 1.31%, 1.73%, and 0.88% respectively, indicating that the test solution is basically stable within 24 h at room temperature.

[0124] (8) Spiked recovery test:

[0125] Take 6 portions of the same batch of samples with known content of the target components, 1 ml for each portion, and add reference substance solutions with a content ratio of approximately 1:1 to each target component. Prepare the test sample solutions according to the above requirements, and determine them under the above chromatographic conditions. Calculate the spiked recovery rate and its RSD value. The results are shown in Table 2. The average spiked recovery rates of baicalin, ephedrine, paeoniflorin, amygdalin, and pseudoephedrine are between 96% and 103%, indicating that this method has good accuracy.

[0126] Table 2 Spiked recovery test results (n = 6)

[0127]

[0128] (9) Specificity investigation

[0129] Take the test sample solution, negative test sample solution, and mixed reference substance solution prepared above, and inject them for detection under the above chromatographic conditions. The results are as Figure 13 shown. The resolution between each target component and its adjacent components is >1.5, and the negative results show no interference, indicating that the target components determined by this method have good specificity.

[0130] (10) Determination of sample content:

[0131] Take 10 batches of samples, prepare the sample solutions according to the above requirements, and determine them under the above chromatographic conditions. The determination results of the contents of ephedrine, pseudoephedrine, amygdalin, paeoniflorin, and baicalin are shown in Table 3.

[0132] Table 3 Determination results of target component contents (n = 10)

[0133]

[0134]

[0135] Example 4 Multicenter clinical study of Xiao'er Jiangqi Pingxiao Mixture based on Zelen design

[0136] (1) Case selection

[0137] The cases were children who visited the Affiliated Hospital of Nanjing University of Chinese Medicine (Jiangsu Provincial Hospital of Chinese Medicine), Yancheng Hospital of Chinese Medicine, and Zhangjiagang Hospital of Chinese Medicine from August 2023 to December 2024 and met the diagnostic criteria for bronchial asthma.

[0138] (2) Group treatment plan

[0139] 1) Western medicine group

[0140] For patients with body weight < 20 kg: 2 ml of budesonide suspension, 1 ml of terbutaline sulfate nebulization solution, and 1 ml of ipratropium bromide solution;

[0141] For patients with body weight ≥ 20 kg: 2 ml of budesonide suspension, 2 ml of terbutaline sulfate nebulization solution, and 2 ml of ipratropium bromide solution;

[0142] Add them to an air compressor nebulizer for aerosol inhalation, 15 minutes each time, twice a day.

[0143] 2) Traditional Chinese medicine group

[0144] Oral administration of Xiao'er Jiangqi Pingxiao Mixture prepared according to Example 1, 20 ml once for children aged 3 - 6 years old, 25 ml once for children over 6 years old, three times a day.

[0145] 3) Integrated traditional Chinese and Western medicine group

[0146] The treatment plan of the above Western medicine group combined with the traditional Chinese medicine group.

[0147] 4) The above plan is continuously treated for 7 days as one course of treatment, and a total of one course of treatment is carried out.

[0148] (3) Research results

[0149] A total of 326 children with asthma attack were included, including 200 males and 126 females. There were 60 cases in the Western medicine group, 168 cases in the traditional Chinese medicine group, and 98 cases in the integrated traditional Chinese and Western medicine group. There was no significant difference in the baseline comparison of the gender and age of the children among the three groups (p > 0.05).

[0150] Table 4 Baseline statistical table

[0151]

[0152] Compare the differences in the disappearance time of the main clinical symptoms of asthma, such as wheezing, gasping, coughing, sputum, and chest tightness, among the three treatment plans. As shown in Table 5 below, Xiao'er Jiangqi Pingxiao Mixture has an advantage in improving expectoration in children, and can shorten the duration of expectoration, and the difference is statistically significant (χ 2 =9.159, P=0.010). The results of multiple comparison analysis: There was a statistically significant difference in sputum between the "traditional Chinese medicine" and "Western medicine" groups (P = 0.026).

[0153] Table 5 Comparison of the basic disappearance time among the main symptoms after treatment

[0154] Wheezing Dyspnea Cough Sputum Chest distress Statistic <![CDATA[χ 2 = 0.221 1 > <![CDATA[χ 2 = 1.773 1 > <![CDATA[χ 2 = 2.092 1 > <![CDATA[χ 2 = 9.159 1 > <![CDATA[χ 2 = 0.677 1 > P value 0.895 0.412 0.351 0.01 0.713

[0155] Note: 1. Kruskal - Wallis H test.

[0156] In terms of within-group comparison, as shown in Table 6, the data distributions of the main symptoms of wheezing, gasping, coughing, sputum, and chest tightness in the traditional Chinese medicine group at the follow-up visit were different from those at the initial visit, and the differences were statistically significant.

[0157] Table 6 Comparison Results of the Main Symptoms of the Traditional Chinese Medicine Group at the Initial and Follow-up Visits

[0158] Wheezing Dyspnea Cough Sputum Chest distress Statistic <![CDATA[Z = 7.368 1 > <![CDATA[Z = 8.099 1 > <![CDATA[Z = 13.561 1 > <![CDATA[Z = 7.899 1 > <![CDATA[Z = 4.226 1 > P value <0.001 <0.001 <0.001 <0.001 <0.001

[0159] Note: 1. Mann Whitney U test; 2. Continuity-corrected χ 2 test.

[0160] As shown in Table 7, the data distributions of the secondary symptoms of nasal itching, sneezing, nasal congestion, runny nose, dry mouth / body fluid, sleep at night, bowel movement, tongue quality, and tongue coating in the traditional Chinese medicine group at the follow-up visit were different from those at the initial visit, and the differences were statistically significant.

[0161] Table 7 Comparison Results of the Secondary Symptoms of the Traditional Chinese Medicine Group at the Initial and Follow-up Visits

[0162] Statistic P value Statistic P value Chest distress <![CDATA[Z = 4.226 1 > <0.001 Aversion to cold <![CDATA[χ 2 = 1.007 2 > 0.316 Nasal itching <![CDATA[Z = 3.757 1 > <0.001 Sleep <![CDATA[Z = 3.222 1 > 0.001 Sneeze <![CDATA[Z = 2.316 1 > 0.021 Urine <![CDATA[Z = 1.293 1 > 0.196 Nasal congestion <![CDATA[Z = 5.887 1 > <0.001 Defecation <![CDATA[Z = 3.978 1 > <0.001 Runny nose <![CDATA[Z = 6.091 1 > <0.001 Tongue body <![CDATA[Z = 6.879 1 > <0.001 Dry mouth / Body fluid <![CDATA[Z = 1.985 1 > 0.047 Tongue coating <![CDATA[Z = 2.436 1 > 0.015 Sore throat <![CDATA[Z = 1.021 1 > 0.307

[0163] Note: 1. Mann Whitney U test; 2. Continuity-corrected χ 2 test.

[0164] The comparison results before and after treatment in the integrated traditional Chinese and Western medicine group are shown in Table 8. The data distributions of the main symptoms of wheezing, gasping, coughing, sputum, and chest tightness at the follow-up visit were different from those at the initial visit, and the differences were statistically significant.

[0165] Table 8 Comparison Results of the Main Symptoms of the Integrated Traditional Chinese and Western Medicine Group at the Initial and Follow-up Visits

[0166] Wheezing Dyspnea Cough Sputum Chest distress Statistic <![CDATA[Z = 5.457 1 > <![CDATA[Z = 6.909 1 > <![CDATA[Z = 10.374 1 > <![CDATA[Z = 6.865 1 > <![CDATA[Z = 3.287 1 > P value <0.001 <0.001 <0.001 <0.001 0.001

[0167] Note: 1. Mann Whitney U test; 2. Pearson χ 2 test.

[0168] As shown in Table 9, after treatment in the integrated traditional Chinese and Western medicine group, the data distributions of the secondary symptoms of nasal itching, nasal congestion, runny nose, sleep at night, bowel movement, and tongue quality at the follow-up visit were different from those at the initial visit, and the differences were statistically significant.

[0169] Table 9 Comparison Results of the Secondary Symptoms of the Integrated Traditional Chinese and Western Medicine Group at the Initial and Follow-up Visits

[0170] Statistic P value Statistic P value Chest distress <![CDATA[Z = 3.287 1 > 0.001 Aversion to cold <![CDATA[Z = 1.092 1 > 0.275 Nasal itching <![CDATA[Z = 2.913 1 > 0.004 Sleep <![CDATA[Z = 3.126 1 > 0.002 Sneeze <![CDATA[Z = 1.841 1 > 0.066 Urine <![CDATA[Z = 1.133 1 > 0.257 Nasal congestion <![CDATA[Z = 5.043 1 > <0.001 Defecation <![CDATA[Z = 3.410 1 > 0.001 Runny nose <![CDATA[Z = 4.850 1 > <0.001 Tongue body <![CDATA[Z = 4.539 1 > <0.001 Dry mouth / Body fluid <![CDATA[χ 2 = 0.619 2 > 0.432 Tongue coating <![CDATA[Z = 1.114 1 > 0.265 Sore throat <![CDATA[χ 2 = 0.008 2 > 0.93

[0171] Note: 1. Mann Whitney U test; 2. Pearson χ 2 test.

[0172] The clinical trial results show that: the Xiao'er Jiangqi Pingxiao Mixture used alone or in combination with the western medicine group has advantages in expectoration compared with the western medicine group, and can shorten the duration of symptoms in children. In terms of the improvement of the main symptoms and secondary symptoms, the Xiao'er Jiangqi Pingxiao Mixture used alone or in combination with the western medicine group has definite curative effects on the five main symptoms during the asthma attack period, and also has a certain improvement effect on each secondary symptom.

[0173] Although the embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A pediatric qi-descending and asthma-calming mixture, characterized in that, It is prepared by the following method: (1) Take 1 - 8 parts by weight of honey - processed Ephedra, 6 - 14 parts of blanched bitter apricot seeds, 8 - 16 parts of stir - fried white peony roots, 6 - 14 parts of uncaria rhynchophylla, 6 - 14 parts of perilla seeds, 6 - 14 parts of plantain seeds, 6 - 14 parts of radish seeds, and 4 - 12 parts of scutellaria baicalensis. Place them in an extraction tank, soak with water, decoct and extract multiple times, filter, combine the filtrates, take an appropriate amount of sucrose, dissolve it in water, mix it with the filtrate, and concentrate under reduced pressure; (2) Centrifuge the concentrated liquid obtained in step (1) with a straight - tube high - speed centrifuge to obtain the mixture.

2. The pediatric qi-lowering and asthma-calming mixture according to claim 1, characterized in that, It is prepared by the following method: Take 1 - 8 parts by weight of honey - processed Ephedra, 6 - 14 parts of blanched bitter apricot seeds, 8 - 16 parts of stir - fried white peony roots, 6 - 14 parts of uncaria rhynchophylla, 6 - 14 parts of perilla seeds, 6 - 14 parts of plantain seeds, 6 - 14 parts of radish seeds, and 4 - 12 parts of scutellaria baicalensis. Place them in an extraction tank, add 10 times the amount of water for the first time, soak for 0.5 h, decoct for 0.5 h, and filter; add 8 times the amount of water for the second time, decoct for 0.5 h, and filter; take an appropriate amount of sucrose, dissolve it in water, combine it with the two filtrates, concentrate under reduced pressure to a relative density of about 1.07 at 60 °C, place it in a straight - tube high - speed centrifuge, and centrifuge at a rotation speed of 16000 r / min and a flow rate of 4 - 6 L / min to obtain the mixture.

3. A quality inspection method for the Pediatric Qi-Dropping and Wheezing-Relieving Mixture prepared by the preparation method of claim 1 or 2, characterized in that, It includes thin - layer chromatography qualitative detection and high - performance liquid chromatography quantitative detection.

4. The quality inspection method according to claim 3, characterized in that, The thin - layer chromatography qualitative detection includes the following steps: (1) Identification of Ephedra by thin - layer chromatography, the specific steps are as follows: 1) Take the mixture, add a few drops of concentrated ammonia, extract with dichloromethane, combine the dichloromethane layers, evaporate to dryness in a water bath, dissolve the residue in methanol to obtain the test solution; take the traditional Chinese medicine compound composition without Ephedra, and prepare the negative solution without Ephedra by the same method as in claim 1; 2) Take another Ephedra control medicinal material powder, add a few drops of concentrated ammonia test solution, then add chloroform, heat under reflux, filter, evaporate the filtrate to dryness, and dissolve the residue in methanol to obtain the Ephedra control medicinal material solution; 3) Absorb the above three solutions and spot them on the same silica gel G thin - layer plate respectively. Use chloroform - methanol - concentrated ammonia as the developing agent, develop, take out, dry in air, spray with ninhydrin test solution, and heat until the spots are clearly colored; in the chromatogram of the test solution, at the corresponding position of the chromatogram of the control medicinal material, there are spots of the same color, and there is no interference from the negative. (2) Identification of bitter apricot seeds by thin - layer chromatography, the specific steps are as follows: 1) Take the mixture, add anhydrous ethanol, let it stand, filter, evaporate the filtrate to dryness, dissolve it in water, extract it with water - saturated n - butanol in several times, combine the n - butanol layers, evaporate to dryness, dissolve the residue in methanol to obtain the test solution; take the traditional Chinese medicine compound composition without blanched bitter apricot seeds, and prepare the negative solution without blanched bitter apricot seeds by the same method as in claim 1; 2) Take another amygdalin reference substance, dissolve it in methanol to prepare the amygdalin reference substance solution; 3) Absorb the above three solutions and spot them on the same silica gel GF254 thin - layer plate respectively. Use ethyl acetate - methanol - water as the developing agent, develop, take out, dry in air, spray with anisaldehyde - sulfuric acid color - developing agent solution, and heat until the spots are clearly colored; in the chromatogram of the test solution, at the corresponding position of the chromatogram of the reference substance solution, there are spots of the same color, and there is no interference from the negative. (3) Identification of white peony roots by thin - layer chromatography, the specific steps are as follows: 1) taking the mixture, evaporating it to dryness, adding ethanol for ultrasonic treatment, filtering, evaporating the filtrate to dryness, dissolving the residue in ethanol, and using it as the test solution; taking the Chinese herbal compound composition lacking white peony root, and preparing a white peony root negative solution according to the same method as claimed in claim 1; 2) Separately, obtain white peony root control medicinal material powder, add ethanol to shake, filter, evaporate the filtrate to dryness, and dissolve the residue in ethanol to prepare the white peony root control medicinal material test solution; 3) Take the paeoniflorin reference substance and add ethanol to prepare a paeoniflorin reference substance solution; 4) Pipette the three solutions above and spot them separately on the same silica gel G thin layer plate. Develop with chloroform-ethyl acetate-methanol-formic acid as the developing solvent. Remove the plate, dry it in air, spray it with vanillin sulfuric acid solution, and heat it until the spots are clearly colored. If the chromatogram of the test solution shows spots of the same color at the corresponding positions in the chromatograms of the control medicinal material and the reference substance solution, no interference is observed in the negative test. (4) Thin layer chromatography identification of Scutellaria baicalensis, the specific steps are as follows: 1) Take the mixture, adjust the pH to 2-3 with dilute hydrochloric acid, extract with ethyl acetate by shaking, combine the two ethyl acetate extracts, evaporate to dryness, dissolve the residue in methanol, and use it as the test solution; take the traditional Chinese medicine compound composition lacking Scutellaria baicalensis, and prepare a Scutellaria baicalensis negative solution according to the same method as claim 1; 2) Separately, take a reference herb of Scutellaria baicalensis, add ethyl acetate-methanol, heat to reflux, cool, filter, evaporate the filtrate to dryness, dissolve the residue in methanol, and use the supernatant as the reference herb solution of Scutellaria baicalensis; 3) Pipette the above three solutions and spot them on the same polyamide film respectively. Use toluene-ethyl acetate-methanol-formic acid as the developing agent, develop, take out, dry, spray with aluminum chloride solution, dry, and examine under ultraviolet light. In the chromatogram of the test solution, at the corresponding position in the chromatogram of the control medicinal material solution, spots of the same color appear under fluorescence, indicating negative and no interference.

5. The quality inspection method according to claim 4, characterized in that Step (1) thin layer chromatography identification of Ephedra using chloroform-methanol-concentrated ammonia in a volume ratio of 20:5:0.5 as a developing solvent; Step (2) identification by thin layer chromatography of bitter almonds using ethyl acetate-methanol-water in a volume ratio of 8:1:1 as the developing solvent; Step (3) thin layer chromatography identification of white peony root, using chloroform-ethyl acetate-methanol-formic acid in a volume ratio of 40:5:10:0.2 as a developing solvent; Step (4) identification of Scutellaria baicalensis by thin layer chromatography using toluene-ethyl acetate-methanol-formic acid in a volume ratio of 10:3:1:2 as a developing solvent.

6. The quality inspection method according to claim 3, wherein The high performance liquid phase quantitative detection comprises the following steps: (1) Preparation of reference solution: accurately weigh ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride reference substances, dissolve in methanol, sonicate, adjust to volume, and shake well to prepare reference solution; (2) Preparation of test solution: accurately measure the mixture, add 70% methanol to the volume, ultrasonically extract, weigh the weight, make up the weight with 70% methanol, shake well, centrifuge, take the supernatant, filter, and obtain; (3) Drawing of standard curve: Take the reference solution prepared in step (1) above, dilute it step by step with methanol, accurately aspirate and inject it into a high performance liquid chromatograph for determination; use the mass concentration X of the reference solution as the abscissa and the peak area Y as the ordinate to obtain the regression equations of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride; (4) Content determination: Precisely pipette the reference solution in step (1) and the test solution in step (2) into the liquid chromatograph respectively, and calculate the contents of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride in the test solution according to the standard curve equation in step (3).

7. The quality inspection method according to claim 6, characterized in that Step (1) Preparation of reference substance solution: Weigh accurately reference substances of ephedrine hydrochloride, baicalin, amygdalin, paeoniflorin, and pseudoephedrine hydrochloride, dissolve them with methanol, and prepare single reference substance solutions with mass concentrations of 1.01, 0.89, 1.45, 0.94, and 0.97 mg·ml -1 respectively. Accurately pipette appropriate amounts of the above-mentioned reference substance solutions into a 20-ml volumetric flask, make up the volume with methanol, and prepare a mixed reference substance solution with mass concentrations of 0.051, 0.134, 0.058, 0.047, and 0.048 mg·ml -1 respectively.

8. The quality inspection method according to claim 6, wherein (2) Preparation of the test solution: Precisely measure 2 ml of the mixture, add 70% methanol to make up the volume to a 10-ml volumetric flask, perform ultrasonic extraction at a power of 100 W and a frequency of 40 KHz for 30 min, weigh, make up the weight with 70% methanol, shake well, centrifuge at 3000 r / min for 10 min, take the supernatant, and filter to obtain the solution.

9. The quality inspection method according to claim 6, wherein Step (3) Drawing of the standard curve: Take the reference substance solution in the above step (1), and serially dilute it with methanol to obtain 5 concentrations. Precisely pipette and inject 10 μL respectively, and inject into a high performance liquid chromatograph for determination; with the mass concentration X of the reference substance as the abscissa and the peak area Y as the ordinate, the regression equation of ephedrine hydrochloride is Y = 16.17X + 8.5526, R 2 = 0.9999, the regression equation of pseudoephedrine hydrochloride is Y = 16.691X + 8.941, R 2 = 0.9999, the regression equation of baicalin is Y = 33.302X + 38.182, R 2 = 0.9994, the regression equation of amygdalin is Y = 11.945X + 3.6842, R 2 = 0.9994, the regression equation of paeoniflorin is Y = 3.6332X + 0.9838, R 2 = 0.9996.

10. The quality inspection method according to claim 6, characterized in that, The chromatographic conditions for step (4) are as follows: chromatographic column Agilent SB-Phenyl, with a specification of 4.6 mm × 250 mm, 5 μm; mobile phase: acetonitrile as phase A, 0.2% phosphoric acid aqueous solution as phase B; gradient elution: 0 - 11 min, 5% → 5% A; 11 - 12 min, 5% → 8% A; 12 - 15 min, 8% → 8% A; 15 - 26 min, 8% → 20% A; 26 - 30 min, 20% → 23% A; 30 - 38 min, 23% → 40% A; column temperature 25°C; detection wavelength 210 nm; flow rate 1 ml·min -1 ; injection volume 10 μL.

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