A quality control method and application of Baibei Yifei capsules

Abstract

The present invention relates to a quality control method and application of Baibei Yifei capsules, belonging to the field of biomedicine technology. The quality control method includes raw material control, preparation method control, identification method and content determination method. The present invention performs quality management on the whole process of product production, and realizes the safety, effectiveness and controllability of the medicine. The identification items are increased to 6 items, and the identification method of Panax notoginseng is improved, which is 3 more than the original standard, and the TLC method identification of licorice, Bletilla striata and Fritillaria thunbergii is newly added; at the same time, when determining the content, the content of ginsenoside Rg1 is increased from not less than 0.4 mg / grain of the current standard to not less than 0.6 mg / grain, and the increase is 50%, with higher and better accuracy. Clinical studies have shown that Baibei Yifei capsules have good effects on the treatment of chronic obstructive pulmonary disease (COPD) and tuberculosis, and can achieve the effect of reducing toxicity and increasing efficacy when combined with anti-tuberculosis drugs, and are easy to promote and apply.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a quality control method and application of Baibei Yifei Capsules. Background Art

[0002] Baibei Yifei Capsule was developed by Kunming Qunfang Pharmaceutical Co., Ltd., which was acquired by Yunnan Yongzitang Pharmaceutical Co., Ltd. It is an optimized product of the patent "a drug for treating lung cough and bronchitis" (patentee: Zhang Qiongfang, patent number: ZL200310121018.9). It is a Yi medicine that has been on the market for many years and is now owned and produced by Yunnan Yongzitang Pharmaceutical Co., Ltd. (National Medicine Standard Z20025124, Implementation Standard WS-10115 (ZD-0115)-2002-2012Z). It is composed of 10 herbs: Stemona, Lilium, Fritillaria, Platycodon, Aster, Mahonia, Bletilla, Pumice, Panax notoginseng, and Licorice. It has the effects of nourishing yin and activating blood circulation, relieving cough and reducing phlegm. It is used to treat chronic cough caused by insufficient lung yin, as well as chronic cough caused by bronchitis and tuberculosis. It has been used clinically for a long time with good results.

[0003] However, Chinese medicine is extensive and profound, and it is obviously a historical responsibility to conduct in-depth research, tap its potential, and carry forward Chinese medicine. According to the theory of Chinese medicine, this medicine, in its essence, still has many values ​​​​worth exploring and developing. If the application (effect) of Baibei Yifei Capsule can be expanded to the treatment of tuberculosis and COPD, it will be a great achievement.

[0004] The lesions of pulmonary tuberculosis are mainly in the lungs, with lung yin deficiency as the main lesion. According to its pathological characteristics of "mainly balancing yin deficiency", the most common TCM syndrome type in clinical practice is lung yin deficiency syndrome, which is clinically manifested as dry cough with little sputum, blood in sputum or hemoptysis, short cough, hot hands and feet, and dry mouth and tongue. Chronic obstructive pulmonary disease (COPD), referred to as COPD, is a common disease that can be prevented and treated. COPD is clinically characterized by cough, sputum, wheezing, and chest tightness. In recent years, the global morbidity and mortality of the disease have shown a significant upward trend, which not only seriously affects the quality of life of patients, but also affects their ability to work to a certain extent. In recent years, TCM has shown significant effects on the prevention and treatment of COPD. Clinical observations show that lung yin deficiency is a common syndrome of COPD, whether in the remission period or the acute exacerbation period. According to TCM's understanding of this disease and the corresponding treatment principles and methods, Baibei Yifei Capsules should play a role.

[0005] In the existing drug standards, there are only three identification items, and there are problems with the identification method and content determination of Panax notoginseng. When identifying Panax notoginseng, the thin layer chromatography has serious tailing, and it is difficult to determine whether there are spots of the same color; in the content determination, due to the limitations of its detection method, it is impossible to accurately detect the content of ginsenoside Rg1, so the content for quality control has always been at a low level. In addition, the current national requirements for drug standards require that the identification items cover 1 / 3 to 1 / 2 of the raw materials, so the current drug standards do not meet the relevant national requirements. Therefore, how to overcome the shortcomings of the existing technology is an urgent problem to be solved in the field of biopharmaceutical technology. Summary of the invention

[0006] The purpose of the present invention is to solve the deficiencies of the prior art and to provide a quality control method for Baibei Yifei Capsules and their application in the preparation of drugs for treating COPD and pulmonary tuberculosis.

[0007] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0008] The first aspect of the present invention provides a quality control method for Baibei Yifei Capsules, including raw material control, preparation method control, identification method and content determination method, which are as follows:

[0009] First, raw material control:

[0010] The 1000 single-dose Baibei Yifei capsules are prepared from the following active ingredients in weight ratio:

[0011] Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g;

[0012] The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines;

[0013] Second, the preparation method control:

[0014] Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside;

[0015] Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting;

[0016] Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules;

[0017] Third, identification method:

[0018] A. Microscopic examination:

[0019] Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long;

[0020] B. Thin layer chromatography detection of Panax notoginseng:

[0021] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0022] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0023] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, place overnight, filter, take the filtrate and evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 5-10ul of the test solution and 5ul of the reference substance solution, respectively, and spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0024] D. Thin layer chromatography detection of licorice:

[0025] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to the residue to dissolve, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way; According to the thin layer chromatography test, 2-5ul of the test sample solution and 2ul of the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 is used as the developing solvent. The plate is developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test sample, a main spot of the same color appears at the corresponding position of the chromatogram of the control medicinal material;

[0026] E. Thin layer chromatography detection of Bletilla striata:

[0027] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 5-10u1 of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C for 5-10 minutes, and then leave it for 30-60 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0028] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0029] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 5-10u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0030] Fourth, content determination method:

[0031] Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia;

[0032] a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0033] b. Preparation of reference solution: Take ginsenoside Rg1 reference substance and add methanol to prepare a solution containing ginsenoside Rg10.3mg per 1ml;

[0034] c. Preparation of test solution: Take 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the mark, shake well, filter, take the filtrate, and obtain;

[0035] d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine;

[0036] Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

[0037] Furthermore, preferably, in step (1), the particle size of the fine powder is 80-100 mesh; in step (2), filtering is performed using a 80-100 mesh sieve, and the material under the sieve is taken.

[0038] Furthermore, preferably, in step (3), the sieving is through a 60-mesh sieve, and the sieve residue is taken; and the drying temperature is 70-80°C.

[0039] Furthermore, preferably, in step c of the content determination method, the inner diameter of the D101 macroporous adsorption resin column is 2 cm and the column length is 15 cm.

[0040] A second aspect of the present invention provides the use of Baibei Yifei Capsules obtained by the above-mentioned Baibei Yifei Capsules quality control method in the preparation of drugs for treating chronic obstructive pulmonary disease.

[0041] A third aspect of the present invention provides the use of Baibei Yifei Capsules obtained by the above-mentioned Baibei Yifei Capsules quality control method in the preparation of drugs for treating pulmonary fibrosis.

[0042] A fourth aspect of the present invention provides the use of Baibei Yifei Capsules obtained by the above-mentioned Baibei Yifei Capsules quality control method in the preparation of drugs for treating pulmonary tuberculosis.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] 1. Carry out quality management on the whole process of product production to ensure the safety, effectiveness and controllability of drugs.

[0045] 2. The number of identification items has been increased to 6, and the identification method of Panax notoginseng has been improved, with 3 more items added than the original standard. The TLC method for identification of licorice, white schizonepeta and Fritillaria thunbergii has been added, accounting for 1 / 3-1 / 2 of the total prescription that meets the national requirements.

[0046] 3. The content determination is a newly created method, which increases the content of ginsenoside Rg1 from the current standard of no less than 0.4 mg / grain to no less than 0.6 mg / grain, an increase of 50%, with higher and better accuracy.

[0047] 4. Pharmacological studies have shown that:

[0048] (1) Baibei Yifei Capsules improve lung function and lung tissue pathological changes in rats with chronic obstructive pulmonary disease (COPD) induced by endotoxin and smoke.

[0049] (2) Baibei Yifei Capsule has an improving effect on bleomycin-induced pulmonary fibrosis.

[0050] (3) Reducing inflammatory cells, lowering IL-8, IL-17, ET-1 and MDA levels, and increasing SOD and NO levels may be one of the mechanisms of action.

[0051] (4) Baibei Yifei Capsules have antibacterial, anti-inflammatory, antitussive, antiasthmatic and expectorant effects.

[0052] 5. Clinical studies show that:

[0053] (1) Baibei Yifei Capsule combined with conventional anti-tuberculosis treatment is better than conventional basic treatment alone, safe to use, and effective in reducing the toxic and side effects of conventional Western medicine treatment. The product of the present invention can achieve the effect of reducing toxicity and increasing efficacy when combined with anti-tuberculosis drug treatment:

[0054] a. Evaluation of the therapeutic effect of chest imaging lesion absorption

[0055] The total absorption rate of the treatment group was 87.10%; the total effective rate of the control group was 63.64%. The comparison of chest imaging efficacy between the two groups was statistically significant (P < 0.05). This shows that the lesion absorption rate of the treatment group was greater than that of the control group, and both treatments had therapeutic effects, and the treatment group was better than the control group.

[0056] b. Evaluation of sputum smear or sputum culture results

[0057] The sputum smear or sputum culture results of the two groups of patients before and after treatment were compared within the group, and the results were statistically significant (P value < 0.01), indicating that both treatment regimens had therapeutic effects.

[0058] The sputum smear or sputum culture results of the treatment group and the control group before treatment were compared, and the results were not statistically significant (P value>0.05), indicating that there was no significant difference in the sputum smear or sputum culture results of the two groups of patients before treatment.

[0059] The sputum smear or sputum culture results of the two groups of patients after treatment were compared, and there was a statistical difference (P<0.05). The sputum smear or sputum culture results of the treatment group were lower than those of the control group after treatment. The negative conversion rate of sputum bacteria: the negative conversion rate of the treatment group was 80.00%; the negative conversion rate of the control group was 42.86%. This shows that the treatment effect of the treatment group is better than that of the control group.

[0060] c. Evaluation of TCM syndrome accumulation

[0061] The total effective rate of the treatment group was 96.77%, and the cure rate was 9.68%; the total effective rate of the control group was 69.70%, and the cure rate was 3.03%. The efficacy of the treatment group was better than that of the control group, with statistical significance (P<0.05).

[0062] The TCM syndrome scores of the treatment group and the control group after treatment were compared within the group, and the results showed significant statistical differences (P < 0.01), indicating that both treatment plans can improve TCM syndrome symptoms.

[0063] The TCM syndrome scores of the treatment group and the control group before treatment were compared between the two groups, and the results were not statistically significant (P>0.05), indicating that there was no difference before treatment. The TCM syndrome scores of the treatment group and the control group after treatment were compared between the two groups, and the results were statistically significant (P<0.05). And by comparing the TCM syndrome scores of the two groups of patients, it was found that the scores of the patients in the treatment group after treatment were lower than those in the control group, which shows that the treatment plan of the treatment group is more advantageous than that of the control group.

[0064] d. Safety evaluation

[0065] The UA of the patients in the treatment group and the control group increased (P < 0.05), and the degree of increase of UA in the treatment group after the treatment was lower than that in the control group (P < 0.05), indicating that anti-tuberculosis Western medicine has the side effect of increasing the patient's blood uric acid, and Baibei Yifei Capsule can reduce the patient's blood uric acid during the treatment process; the abnormalities of ALT and AST in the two groups of patients were statistically different after treatment (P < 0.05), indicating that Baibei Yifei Capsule has a more obvious effect of protecting the liver and reducing enzymes during the treatment course; the number of gastrointestinal symptoms in the two groups of patients was statistically different after treatment (P < 0.05), indicating that Baibei Yifei Capsule is effective in alleviating the gastrointestinal symptoms of patients during the treatment course; and the number of mild allergic reaction symptoms was not statistically significant after treatment (P > 0.05), indicating that Baibei Yifei Capsule has no obvious effect on alleviating mild allergic reactions in patients during the treatment course. The other safety indicators were all within the normal range.

[0066] (2) Baibei Yifei Capsule has the effect of improving TCM symptoms in the treatment of chronic obstructive pulmonary disease (COPD) with lung yin deficiency, and its efficacy in treating symptoms such as cough, wheezing, chest tightness and shortness of breath is better than that of the control group:

[0067] a. The total score of TCM syndrome and the scores of each main and secondary symptom of TCM in the experimental group and the control group before and after treatment were compared within the group. The results showed that the differences were significant (P < 0.01), indicating that both groups of drugs had obvious therapeutic effects.

[0068] b. In terms of improving the efficacy of TCM syndrome, the effective rate of the experimental group after treatment was 83.75%, and the effective rate of the control group was 75.0%. After the rank sum test, the difference between the experimental group and the control group was statistically significant (P < 0.05), indicating that the experimental group was superior to the control group in improving the efficacy of TCM syndrome.

[0069] c. Comparison of changes in the scores of individual TCM symptoms between the two groups of patients before and after treatment: the experimental group was significantly better than the control group in improving cough and asthma symptoms (P < 0.01); the experimental group was better than the control group in improving chest tightness and shortness of breath symptoms (P < 0.05), while there was no significant difference in improving sputum color and dry mouth and throat symptoms between the two groups (P > 0.05).

[0070] d. It is safe for clinical use, and no obvious adverse reactions or toxic side effects have been found. BRIEF DESCRIPTION OF THE DRAWINGS

[0071] Figure 1 This is a flow chart of the preparation method of Baibei Yifei Capsule of the present invention;

[0072] Figure 2It is a TLC chromatogram of Panax notoginseng; wherein, 1. notoginseng saponin R1, ginsenoside Rg, mixed reference solution; 2. notoginseng saponin R, reference solution; 3. ginsenoside Rg1 reference solution; 4-13. Baibei Yifei capsule test solution (batch numbers are: 20151007, 2015051, 20160504, 20150201, 20150504, 20150708, 20151206, 20160109, 20160306, 20160521);

[0073] Figure 3 It is a TLC chromatogram of Fritillaria thunbergii; wherein, 1-10. Baibei Yifei Capsules (batch numbers are: 20151007, 20150512, 20160504, 20150201, 20150504, 20150708, 20151206, 20160109, 20160306, 20160521); 11. Fritillaria sucrase A reference solution; 12. Fritillaria sucrase B reference solution;

[0074] Figure 4 It is a TLC chromatogram of licorice; wherein, 1-10, 12-14. Baibei Yifei capsule test solution (batch numbers are: 20151007, 20150512, 20160504, 20150201, 20150504, 20150708, 20151206, 20160109, 20160306, 20160521, 20160306, 20150520, 20160206; 11, 15. licorice control medicinal material solution;

[0075] Figure 5 The TLC chromatogram of Bletilla striata; wherein, 1, 15 are Bletilla striata medicinal materials; 2, 8, 14 are Bletilla striata control medicinal materials; 3-7, 9-13 are Baibei Yifei capsules (batch numbers: 20160521, 20150708, 20170105, 20160109, 20151007, 20160504, 20150504, 20150201, 20160520, 20161206);

[0076] Figure 6 The effect of Baibei Yifei capsule on pulmonary histopathology of COPD rats induced by endotoxin (200×);

[0077] Figure 7 The effect of Baibei Yifei capsule on pulmonary artery pathology in rats with COPD induced by endotoxin (200×);

[0078] Figure 8The effect of Baibei Yifei capsule on lung histopathology of smoke-induced COPD rats (200×);

[0079] Fig. 9 The effect of Baibei Yifei capsule on pulmonary artery pathology in smoke-induced COPD rats (200×);

[0080] Fig.10 The effect of Baibei Yifei capsule on the pathological changes of lung tissue in rats with pulmonary fibrosis induced by bleomycin (200×);

[0081] Fig.11 The effect of Baibei Yifei capsule on the Masson staining of lung tissue in rats with pulmonary fibrosis induced by bleomycin (400×);

[0082] Fig.12 Gender of the two groups of patients;

[0083] Fig.13 The comparison between groups was for the sputum smear or sputum culture results before treatment;

[0084] Fig.14 Comparison of sputum smear or sputum culture results between groups after treatment;

[0085] Fig.15 is the percentage of TCM syndrome efficacy in the treatment group;

[0086] Fig.16 It is the percentage of TCM syndrome efficacy in the control group. DETAILED DESCRIPTION

[0087] The present invention is further described in detail below in conjunction with embodiments.

[0088] Those skilled in the art will appreciate that the following examples are only used to illustrate the present invention and should not be considered to limit the scope of the present invention. If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the art or the product specifications are used. If the manufacturer of the materials or equipment used is not specified, they are all conventional products that can be purchased.

[0089] The reagents and materials used in the present invention are in accordance with the requirements of the pharmacopoeia.

[0090] Example 1

[0091] A quality control method for Baibei Yifei Capsules, including raw material control, preparation method control, identification method and content determination method, is as follows:

[0092] First, raw material control:

[0093] The 1000 single-dose Baibei Yifei capsules are prepared from the following active ingredients in weight ratio:

[0094] Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g;

[0095] The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines;

[0096] Second, the preparation method control:

[0097] Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside;

[0098] Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting;

[0099] Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules;

[0100] Third, identification method:

[0101] A. Microscopic examination:

[0102] Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long;

[0103] B. Thin layer chromatography detection of Panax notoginseng:

[0104] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0105] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0106] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, let it stand overnight, filter, take the filtrate and evaporate it to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 5ul of the test solution and 5ul of the reference substance solution, respectively, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0107] D. Thin layer chromatography detection of licorice:

[0108] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to dissolve the residue, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way According to the thin layer chromatography test, 2ul of the test sample solution and 2ul of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position of the chromatogram of the control medicinal material.

[0109] E. Thin layer chromatography detection of Bletilla striata:

[0110] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 5ul of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for 5 minutes, and then let it stand for 30 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0111] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0112] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 5u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0113] Fourth, content determination method:

[0114] Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia;

[0115] a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0116] b. Preparation of reference solution: Take ginsenoside Rg1 reference substance and add methanol to prepare a solution containing ginsenoside Rg10.3mg per 1ml;

[0117] c. Preparation of test solution: Take 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the mark, shake well, filter, take the filtrate, and obtain;

[0118] d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine;

[0119] Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

[0120] Example 2

[0121] A quality control method for Baibei Yifei Capsules, including raw material control, preparation method control, identification method and content determination method, is as follows:

[0122] First, raw material control:

[0123] The 1000 single-dose Baibei Yifei capsules are prepared from the following active ingredients in weight ratio:

[0124] Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g;

[0125] The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines;

[0126] Second, the preparation method control:

[0127] Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside;

[0128] Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting;

[0129] Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules;

[0130] Third, identification method:

[0131] A. Microscopic examination:

[0132] Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long;

[0133] B. Thin layer chromatography detection of Panax notoginseng:

[0134] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0135] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0136] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, let it stand overnight, filter, take the filtrate and evaporate it to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 5ul of the test solution and 5ul of the reference substance solution, respectively, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0137] D. Thin layer chromatography detection of licorice:

[0138] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to dissolve the residue, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way According to the thin layer chromatography test, 2ul of the test sample solution and 2ul of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position of the chromatogram of the control medicinal material.

[0139] E. Thin layer chromatography detection of Bletilla striata:

[0140] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 5ul of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for 5 minutes, and then let it stand for 30 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0141] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0142] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 5u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0143] Fourth, content determination method:

[0144] Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia;

[0145] a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0146] b. Preparation of reference solution: Take ginsenoside Rg1 reference substance and add methanol to prepare a solution containing ginsenoside Rg10.3mg per 1ml;

[0147] c. Preparation of test solution: Take 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the mark, shake well, filter, take the filtrate, and obtain;

[0148] d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine;

[0149] Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

[0150] In step (1), the particle size of the fine powder is 80 mesh; in step (2), an 80-mesh sieve is used for filtration, and the material under the sieve is taken.

[0151] In step (3), the sieving is through a 60-mesh sieve, and the sieve material is taken; and the drying temperature is 70°C.

[0152] In step c of the content determination method, the inner diameter of the D101 macroporous adsorption resin column is 2 cm and the column length is 15 cm.

[0153] Example 3

[0154] A quality control method for Baibei Yifei Capsules, including raw material control, preparation method control, identification method and content determination method, is as follows:

[0155] First, raw material control:

[0156] The 1000 single-dose Baibei Yifei capsules are prepared from the following active ingredients in weight ratio:

[0157] Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g;

[0158] The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines;

[0159] Second, the preparation method control:

[0160] Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside;

[0161] Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting;

[0162] Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules;

[0163] Third, identification method:

[0164] A. Microscopic examination:

[0165] Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long;

[0166] B. Thin layer chromatography detection of Panax notoginseng:

[0167] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0168] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0169] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, place overnight, filter, take the filtrate and evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 10ul of the test solution and 5ul of the reference substance solution, respectively, and spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0170] D. Thin layer chromatography detection of licorice:

[0171] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to dissolve the residue, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way According to the thin layer chromatography test, 5ul of the test sample solution and 2ul of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position of the chromatogram of the control medicinal material.

[0172] E. Thin layer chromatography detection of Bletilla striata:

[0173] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 10ul of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for 10 minutes, and then leave it for 60 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0174] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0175] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 10u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0176] Fourth, content determination method:

[0177] Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia;

[0178] a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0179] b. Preparation of reference solution: Take ginsenoside Rg1 reference substance and add methanol to prepare a solution containing ginsenoside Rg10.3mg per 1ml;

[0180] c. Preparation of test solution: Take 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the mark, shake well, filter, take the filtrate, and obtain;

[0181] d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine;

[0182] Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

[0183] In step (1), the particle size of the fine powder is 100 mesh; in step (2), a 100-mesh sieve is used for filtration, and the material under the sieve is taken.

[0184] In step (3), the sieving is through a 60-mesh sieve, and the sieve material is taken; and the drying temperature is 80°C.

[0185] In step c of the content determination method, the inner diameter of the D101 macroporous adsorption resin column is 2 cm and the column length is 15 cm.

[0186] Example 4

[0187] A quality control method for Baibei Yifei Capsules, including raw material control, preparation method control, identification method and content determination method, is as follows:

[0188] First, raw material control:

[0189] The 1000 single-dose Baibei Yifei capsules are prepared from the following active ingredients in weight ratio:

[0190] Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g;

[0191] The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines;

[0192] Second, the preparation method control:

[0193] Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside;

[0194] Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting;

[0195] Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules;

[0196] Third, identification method:

[0197] A. Microscopic examination:

[0198] Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long;

[0199] B. Thin layer chromatography detection of Panax notoginseng:

[0200] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0201] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0202] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, place overnight, filter, take the filtrate and evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 8ul of the test solution and 5ul of the reference substance solution, respectively, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0203] D. Thin layer chromatography detection of licorice:

[0204] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to dissolve the residue, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way According to the thin layer chromatography test, 4ul of the test sample solution and 2ul of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position of the chromatogram of the control medicinal material.

[0205] E. Thin layer chromatography detection of Bletilla striata:

[0206] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 8ul of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for 8 minutes, and then leave it for 45 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0207] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0208] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 7u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0209] Fourth, content determination method:

[0210] Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia;

[0211] a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0212] b. Preparation of reference solution: Take ginsenoside Rg1 reference substance and add methanol to prepare a solution containing ginsenoside Rg10.3mg per 1ml;

[0213] c. Preparation of test solution: Take 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the mark, shake well, filter, take the filtrate, and obtain;

[0214] d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine;

[0215] Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

[0216] In step (1), the particle size of the fine powder is 90 mesh; in step (2), a 90-mesh sieve is used for filtration, and the material under the sieve is taken.

[0217] In step (3), the sieving is through a 60-mesh sieve, and the sieve material is taken; and the drying temperature is 75°C.

[0218] In step c of the content determination method, the inner diameter of the D101 macroporous adsorption resin column is 2 cm and the column length is 15 cm.

[0219] Example 5

[0220] 1. Quality Control of Baibei Yifei Capsules

[0221] A quality control method for Baibei Yifei Capsules, including raw material control, preparation method quality control and product quality control. A good medicine should be based on appropriate higher standards to fully guarantee the quality of the medicine; implement GMP, a complete and controllable production process. We firmly believe that the quality of medicines is produced.

[0222] 1. Raw material control

[0223] Prescription of Baibei Yifei Capsule

[0224] Bletilla striata 118g Fritillaria thunbergii 40g Platycodon grandiflorum 67g Stemona 67g

[0225] Lily 67g Pumice 67g Aster 67g Panax notoginseng 40g

[0226] 67g of Chinese magnolia bark and 20g of licorice

[0227] Among them, the nine ingredients of Bletilla striata, Stemona tuber, Aster tataricus, Licorice, Fritillaria thunbergii, Lilium, Panax notoginseng, Platycodon grandiflorum, and Mahonia lactiflora comply with the standards of the Chinese Pharmacopoeia; and the pumice stone complies with the standards of the 2001 edition of "Heilongjiang Province Traditional Chinese Medicine Standards".

[0228] 2. Preparation method control

[0229] (1) Production process flow Figure 1 As shown;

[0230] Weigh the medicinal materials according to the quality ratio of the medicinal materials in the prescription

[0231] Grind Bletilla striata, Pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder and sterilize for later use; add 6 times the amount of water to decoct the six ingredients of Platycodon grandiflorum, Stemona bulb, Lilium bulb, Aster tataricus, Mahonia laurel and Licorice for the first time, add 5 times the amount of water for the second time, and add 4 times the amount of water for the third time, each time for 1 hour, combine the decoctions, filter, and concentrate the filtrate under normal pressure into a thick paste (d≥1.25, room temperature), mix it with the fine powder of Bletilla striata and the other four ingredients, dry, grind, sieve (60 mesh), put it into capsules, and make 1000 capsules.

[0232] (2) The sorting, cleaning, drying, crushing, extraction, concentration, preparation and packaging of medicinal materials are all carried out in a Class D clean area.

[0233] (3) GMP standards are implemented during the production process.

[0234] (4) The prepared capsules should comply with the relevant provisions of the General Chapter 01037 of Part IV of the 2020 edition of the Chinese Pharmacopoeia, and pass the bacterial test and disintegration test.

[0235] 3. Product quality control

[0236] Identification method:

[0237] A. Microscopic examination:

[0238] Take this product and observe it under a microscope: the starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped (Fritillaria thunbergii). Calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long (Bletilla striata).

[0239] B. Thin layer chromatography detection of Panax notoginseng:

[0240] Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the above three solutions are taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product;

[0241] Thin layer chromatography of Panax notoginseng was performed on 10 batches of Baibei Yifei Capsules. The TLC chromatograms are shown in the figure below. Figure 2 As shown;

[0242] C. Thin layer chromatography detection of Fritillaria thunbergii:

[0243] Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, place overnight, filter, take the filtrate and evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of fritillarin A and fritillarin B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 5-10ul of the test solution and 5ul of the reference substance solution, respectively, spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears;

[0244] TLC analysis of Fritillaria thunbergii was performed on 10 batches of Baibei Yifei Capsules. Figure 3 As shown;

[0245] D. Thin layer chromatography detection of licorice:

[0246] Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to the residue to dissolve, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to the residue to dissolve, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way; According to the thin layer chromatography test, 2-5ul of the test sample solution and 2ul of the control medicinal material solution are respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 is used as the developing solvent. The plate is developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test sample, a main spot of the same color appears at the corresponding position of the chromatogram of the control medicinal material;

[0247] TLC analysis of licorice was performed on 10 batches of Baibei Yifei Capsules. The TLC chromatograms are shown in Figure 2. Figure 4 As shown;

[0248] E. Thin layer chromatography detection of Bletilla striata:

[0249] Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, extract twice with ether by shaking, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 5-10u1 of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105°C for 5-10 minutes, and then leave it for 30-60 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material;

[0250] Ten batches of Baibei Yifei capsules were tested for Bletilla striata by thin layer chromatography. The TLC chromatograms are shown in the figure below. Figure 5 As shown;

[0251] F. Thin layer chromatography detection of Platycodon grandiflorum:

[0252] Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 5~1u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears;

[0253] Among them, the thin layer chromatography test refers to the General Rules 0502 of Part IV of the 2020 edition of the Chinese Pharmacopoeia.

[0254] [Inspection] It should comply with the relevant provisions under capsules (General Rules 0103 of Part 4 of the 2020 edition of the Chinese Pharmacopoeia).

[0255] [Content determination] Determined by high performance liquid chromatography (General Rules 0512 of Part IV of the 2020 edition of the Chinese Pharmacopoeia)

[0256] The chromatographic conditions and system suitability test used octadecylsilane bonded silica gel as the filler, acetonitrile-water (18:82) as the mobile phase, and the detection wavelength was 203 nm. The theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000.

[0257] Preparation of reference solution: Take an appropriate amount of ginsenoside Rg1 reference solution, weigh it accurately, and add methanol to prepare a solution containing 0.3 mg of ginsenoside Rg1 per 1 ml.

[0258] Preparation of test solution: Take the contents of 20 tablets of this product, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the scale, shake well, filter, take the filtrate, and obtain;

[0259] The determination method is to accurately pipette 10ul of the reference solution and the test solution respectively, inject them into the liquid chromatograph, and determine the result.

[0260] Each capsule of this product contains ginsenoside Rg1(C 42 H 72 O 14 ) shall not be less than 0.6mg.

[0261] Table 1 Determination results of ginsenoside Rg1 content in Baibei Yifei capsules

[0262] Serial number batch number Sampling volume (g) Content (mg / g) Content (mg / tablet) 1 20150504 2.0017 3.2671 1.0079 2 20150512 2.0155 3.2314 0.9875 3 20150708 2.0032 3.3034 1.0161 4 20151206 2.003 2.6291 0.8126 5 20160109 2.0009 2.6403 0.7768 6 20160306 2.016 2.9544 0.9194 7 20160520 2.0119 2.7136 0.8309 8 20160301 2.0096 2.7277 0.8199 9 20170101 2.0008 2.8924 0.8613 10 20160201 2.0015 3.2559 0.9865

[0263] The average content of ginsenoside Rg1 in 10 batches of Baibei Yifei Capsules was 0.9019 / capsule. Considering that the prescription has many medicinal ingredients and many extraction steps, the content limit is set at 0.60mg / capsule. That is, the content determination limit is: each capsule of this product contains ginsenoside Rg1 (C 42 H 72 O 14 ) shall not be less than 0.6mg.

[0264] 2. Pharmacodynamic trials for experimental chronic obstructive pulmonary disease

[0265] Methods: The COPD model of rats was established by endotoxin (LPS) and fumigation, and the pulmonary fibrosis model of rats was established by intratracheal injection of bleomycin. The lung function was detected by biological signal acquisition system at 2 weeks, 4 weeks and 8 weeks of administration. After 8 weeks of medication, the lung strip tension, total leukocyte count and differential count of bronchoalveolar lavage fluid (BALF) of the animals in the above groups were detected. The contents of SOD, MDA, NO, IL-8, IL-17 and ET-1 in BALF and lung tissue were detected by ELISA. The levels of IgG, IgA and IgM were determined by agar monophasic immunodiffusion. HE staining was used for pathological observation of lung tissue and pulmonary vascular, and Masson staining was used for pathological detection of pulmonary fibrosis. The anti-inflammatory effect was observed by xylene-induced ear swelling in mice and cotton ball granuloma in rats; the minimum inhibitory concentration of the drug was detected by modified Roche medium for tuberculosis, LB nutrient agar medium for Klebsiella pneumoniae, brain heart infusion broth for Streptococcus pneumoniae, and nutrient broth agar medium for Pseudomonas aeruginosa and Staphylococcus aureus, as shown in Table 22. The effects on cough were evaluated by concentrated ammonia, citric acid and electric stimulation of cat superior laryngeal nerve to induce cough; the effects on wheezing were detected by acetylcholine and histamine mixed solution spray method, and the tension of guinea pig isolated tracheal smooth muscle was detected by biological signal acquisition system. The effects of drugs on sputum volume were detected by tracheal segment phenol red method and capillary method.

[0266] result:

[0267] 1. Effect of Baibei Yifei Capsule on COPD in Rats Induced by LPS

[0268] (1) Effects on pulmonary function of rats with COPD induced by endotoxin: 2-8 weeks after modeling, the respiratory rate, tidal volume, peak respiratory flow, and respiratory resistance of the endotoxin model group were significantly increased ( b P<0.01). Compared with the model group at the same time point, 125mg / kg Baibei Yifei capsule for 8 weeks, 250mg / kg for 4-8 weeks, and 500mg / kg Baibei Yifei capsule, 1mg / kg dexamethasone and 250mg / kg Lifei tablets for 2-8 weeks significantly slowed down the respiratory rate, increased the tidal volume and peak respiratory flow, and reduced the respiratory resistance ( c P<0.05, d P<0.01). Compared with the two positive control groups, the effects of the three doses of Baibei Yifei Capsules were weaker after 2-8 weeks of use ( ef P<0.05), as shown in Table 2.

[0269] (2) Effect on lung strip tension in COPD rats: Compared with the model group, the lung strip tension in the 500 mg / kg Baibei Yifei capsule and two positive control drug groups was significantly reduced ( d P<0.01), and the effect strength was comparable, as shown in Table 3.

[0270] (3) Effect on the total and differential white blood cell counts in bronchoalveolar lavage fluid (BALF) of COPD: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect, and 250 mg / kg Baibei Yifei capsule only reduced the total white blood cell count and eosinophil count, but was weaker than the two positive control drugs. 500 mg / kg Baibei Yifei capsule and the two positive control drugs could significantly reduce the total and differential white blood cell counts ( d P<0.01), as shown in Table 4.

[0271] (4) Effects on antioxidant and inflammatory indices in bronchoalveolar lavage fluid and lung tissue of COPD rats: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect on SOD, MDA and NO levels. 250 mg / kg Baibei Yifei capsule only reduced MDA levels and had no significant effect on SOD and NO. 500 mg / kg Baibei Yifei capsule and the two positive control drugs significantly increased SOD and NO levels and reduced MDA levels ( c P<0.05, d P<0.01).

[0272] Compared with the model group, the three doses of Baibei Yifei capsule significantly reduced the levels of IL-8, IL-17 and ET-1 ( c P<0.05, dP<0.01), but the effects of 125 and 250 mg / kg Baibei Yifei capsules were weaker than those of the two positive control drugs ( ef P<0.05), as shown in Tables 5 and 6.

[0273] (5) Effects on immune proteins in lung tissue of COPD rats: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect, 250 mg / kg group only increased IgG and IgM levels, and 500 mg / kg Baibei Yifei capsule increased IgG, IgA and IgM levels ( c P<0.05, d P<0.01), as shown in Table 7.

[0274] (6) Lung tissue and pulmonary vascular pathology

[0275] Model group: The bronchial lumen was obviously narrowed, and the bronchial ciliated epithelial cells were degenerated, necrotic, and shed. A large number of inflammatory cells were infiltrated around the bronchi. The alveolar structure was disordered, the alveolar wall was thinned or broken, the alveolar cavity was enlarged, and fused into bullae. The pulmonary artery wall was significantly thickened, endothelial cells shed, a large number of inflammatory cells infiltrated, and vascular smooth muscle cells proliferated significantly.

[0276] The high-dose Baibei Yifei capsule group can reduce the degeneration, necrosis and shedding of bronchial ciliated epithelial cells, reduce inflammatory cell infiltration, and improve alveolar structure. It can also improve the pathological changes of pulmonary artery, such as Figure 6 and Figure 7 shown.

[0277] 2. Effect of Baibei Yifei Capsule on COPD in Rats Induced by Smoke

[0278] (1) Effects on lung function of COPD rats: 2-8 weeks after modeling, the respiratory rate of the smoke model group was significantly accelerated, the tidal volume was significantly reduced, the peak respiratory flow value was significantly reduced, and the respiratory resistance was significantly increased ( b P<0.01). 125mg / kg Baibei Yifei Capsule for 2-8 weeks and 250mg / kg for 2-4 weeks had no significant effect on respiratory rate, but 8 weeks of medication slowed down the respiratory rate ( c P<0.01), but weaker than the two positive control drugs. 125 and 250 mg / kg Baibei Yifei capsules had no significant effect on peak respiratory flow, tidal volume and respiratory resistance after 2-8 weeks of medication. 500 mg / kg could improve the above indicators, but the effects were weaker than the two positive control drugs ( ef P<0.05), as shown in Table 8.

[0279] (2) Effect on lung strip tension in COPD rats: Compared with the model group, the lung strip tension in the 250 and 500 mg / kg Baibei Yifei capsule groups and the two positive control groups was significantly reduced (d P<0.01), and the effect strength was comparable, as shown in Table 9.

[0280] (3) Effect on the total and differential white blood cell counts in bronchoalveolar lavage fluid (BALF) of COPD: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect, and 250 mg / kg Baibei Yifei capsule only reduced the total white blood cell count and eosinophil count, but was weaker than the two positive control drugs. 500 mg / kg Baibei Yifei capsule could significantly reduce the total and differential white blood cell counts ( d P<0.01), as shown in Table 10.

[0281] (4) Effects on antioxidant and inflammatory indices in bronchoalveolar lavage fluid and lung tissue of COPD rats: Compared with the model group, 125 and 250 mg / kg Baibei Yifei capsules increased SOD levels, but had no significant effect on MDA and NO. 500 mg / kg Baibei Yifei capsules significantly increased SOD and NO levels and reduced MDA levels ( c P<0.05, d P<0.01).

[0282] Compared with the model group, 250 and 500 mg / kg Baibei Yifei capsules significantly reduced IL-8, IL-17 and ET-1 ( c P<0.05, d P<0.01), but weaker than the two positive control drugs ( ef P<0.05), as shown in Tables 11 and 12.

[0283] (5) Effects on immune proteins in lung tissue of COPD rats: Compared with the model group, the 250 mg / kg group only increased the levels of IgG and IgM, while the 500 mg / kg Baibei Yifei capsule increased the levels of IgG, IgA and IgM ( c P<0.05, d P<0.01), as shown in Table 13.

[0284] (6) Lung tissue and pulmonary vascular pathology

[0285] Model group: The bronchial lumen was obviously narrowed, and the bronchial ciliated epithelial cells were degenerated, necrotic, and shed. A large number of inflammatory cells were infiltrated around the bronchi. The alveolar structure was disordered, the alveolar wall was thinned or broken, the alveolar cavity was enlarged, and fused into bullae. The pulmonary artery wall was significantly thickened, endothelial cells shed, a large number of inflammatory cells infiltrated, and vascular smooth muscle cells proliferated significantly.

[0286] The high-dose Baibei Yifei capsule group can reduce the degeneration, necrosis and shedding of bronchial ciliated epithelial cells, reduce inflammatory cell infiltration, and improve alveolar structure. It can also improve the pathological changes of pulmonary artery, such as Figure 8and Fig. 9 shown.

[0287] 3. Effect of Baibei Yifei Capsule on Bleomycin-induced Pulmonary Fibrosis in Rats

[0288] (1) Effects on lung function in rats with pulmonary fibrosis: 125 mg / kg Baibei Yifei Capsules had no significant effect on the four indicators of lung function, and 250 mg / kg had no significant effect on peak respiratory flow. It could slow down the respiratory rate and respiratory resistance, increase tidal volume, and its effect was weaker than that of the equivalent dose of Lifei Tablets, as shown in Table 14.

[0289] (2) Effect on lung strip tension in rats with pulmonary fibrosis: Compared with the model group, 125 and 250 mg / kg Baibei Yifei capsules had no significant effect on lung strip tension, while the tension in the 500 mg / kg Baibei Yifei capsule group was significantly reduced ( d P<0.01), as shown in Table 15.

[0290] (3) Effect on the total and differential white blood cell counts in bronchoalveolar lavage fluid (BALF) of pulmonary fibrosis: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect, and 250 mg / kg Baibei Yifei capsule only reduced the total white blood cell count and eosinophil count, but was weaker than the two positive control drugs. 500 mg / kg Baibei Yifei capsule significantly reduced the total white blood cell count and differential count ( d P<0.01), as shown in Table 16.

[0291] (4) Effects on antioxidant and inflammatory indices in bronchoalveolar lavage fluid and lung tissue of rats with pulmonary fibrosis: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect on SOD, MDA and NO levels. 250 mg / kg Baibei Yifei capsule only reduced MDA level and increased SOD level, but had no significant effect on NO in lung tissue, which was weaker than the two positive control drugs. 500 mg / kg Baibei Yifei capsule significantly increased SOD and NO levels and reduced MDA level ( c P<0.05, d P<0.01).

[0292] Compared with the model group, the medium and high doses of Baibei Yifei capsules significantly reduced the levels of IL-8, IL-17 and ET-1 ( c P<0.05, d P<0.01), but the effect of 250 mg / kg Baibei Yifei capsule was weaker than that of the two positive control drugs ( ef P<0.05), as shown in Tables 17 and 18.

[0293] (5) Effects on immune proteins in lung tissue of rats with pulmonary fibrosis: Compared with the model group, 125 mg / kg Baibei Yifei capsule had no significant effect, 250 mg / kg group only increased IgG and IgA levels, and had no significant effect on IgM; 500 mg / kg Baibei Yifei capsule increased the levels of IgG, IgA and IgM ( c P<0.05, d P<0.01), as shown in Table 19.

[0294] (6) Lung tissue and pulmonary vascular pathology (HE staining)

[0295] Model group: The bronchial lumen was obviously narrowed, and the bronchial ciliated epithelial cells were degenerated, necrotic, and shed. A large number of inflammatory cells were infiltrated around the bronchi. The alveolar structure was disordered, the alveolar wall was thinned or broken, the alveolar cavity was enlarged, and fused into bullae. The pulmonary artery wall was significantly thickened, endothelial cells shed, a large number of inflammatory cells infiltrated, and vascular smooth muscle cells proliferated significantly.

[0296] The high-dose Baibei Yifei capsule group can reduce the degeneration, necrosis and shedding of bronchial ciliated epithelial cells, reduce inflammatory cell infiltration, and improve alveolar structure. It can also improve the pathological changes of pulmonary artery, such as Fig.10 shown.

[0297] (7) Pulmonary fibrosis (Masson staining)

[0298] In the model group, myofibroblasts proliferated and aggregated in the alveolar cavity and interstitium, and a large number of wide-band and sheet-shaped collagen fibers appeared, showing diffuse fibrosis changes and a large amount of collagen fibers accumulated.

[0299] The muscle fiber hyperplasia and collagen fiber in the 250 and 500 mg / kg Baibei Yifei capsule groups were significantly reduced compared with the model group. Fig.11 shown.

[0300] 4. Effects of Baibei Yifei Capsule on Inflammation

[0301] (1) Effect of xylene on ear swelling in mice: 200 and 400 mg / kg of Baibei Yifei Capsules had no significant effect in inhibiting swelling, while 800 mg / kg had significant anti-inflammatory and detumescent effects, but weaker than aspirin, as shown in Table 20.

[0302] (2) Effect on cotton ball granuloma in rats: Compared with the model group, 250 and 500 mg / kg of Baibei Yifei capsules and the two positive control drugs significantly reduced the weight of granuloma ( a P<0.05, b P<0.01), the effect of 250 mg / kg Baibei Yifei Capsule was weaker than that of the two positive control drugs, as shown in Table 21.

[0303] 5. Effect of Baibei Yifei Capsules on cough: In the models of cough induced by concentrated ammonia in mice, cough induced by citric acid in guinea pigs, and cough induced by electrical stimulation of the superior laryngeal nerve in cats, low doses of Baibei Yifei Capsules had no obvious effect. Medium and high doses could reduce the number of coughs, prolong the latency period, and increase the threshold of electrical stimulation. The effect of the medium dose was weaker than that of Lifei Tablets and codeine, as shown in Tables 23-25.

[0304] 6. Effect on wheezing: In the asthma model induced by spraying a mixture of acetylcholine and histamine, the three doses of Baibei Yifei capsules prolonged the wheezing latency of guinea pigs. The effects of low and medium doses of Baibei Yifei capsules were weaker than those of the positive control drugs aminophylline and Lifei tablets, as shown in Table 26.

[0305] 7. Effect on the tension of isolated guinea pig tracheal strips: Baibei Yifei Capsules inhibited the contraction of guinea pig tracheal strips induced by histamine in a concentration-dependent manner, but was weaker than aminophylline ( c P<0.01), as shown in Table 27.

[0306] 8. Effect on sputum discharge: The results of the phenol red method for mouse tracheal segments showed that the medium and high doses significantly increased the OD value, but were weaker than ammonium chloride and Lifei tablets. The results of the rat capillary method showed that 250 and 500 mg / kg Baibei Yifei capsules increased the length of sputum in the capillary, which was weaker than ammonium chloride, as shown in Table 28 and Table 29.

[0307] in conclusion:

[0308] (1) Baibei Yifei Capsules improve lung function and lung tissue pathological changes in rats with chronic obstructive pulmonary disease (COPD) induced by endotoxin and smoke;

[0309] (2) Baibei Yifei Capsule has an improving effect on bleomycin-induced pulmonary fibrosis;

[0310] (3) Reducing inflammatory cells, lowering IL-8, IL-17, ET-1 and MDA levels, and increasing SOD and NO levels may be one of the mechanisms of action;

[0311] (4) Baibei Yifei Capsules have antibacterial, anti-inflammatory, antitussive, antiasthmatic and expectorant effects;

[0312] (5) Minimum inhibitory concentration in vitro: Baibei Yifei Capsules have a strong inhibitory effect on Streptococcus pneumoniae and Klebsiella pneumoniae in vitro, and have a certain inhibitory effect on Mycobacterium tuberculosis and Staphylococcus aureus.

[0313] Table 2. Effects of Baibei Yifei Capsule on Lung Function in COPD Rats Induced by LPS

[0314]

[0315] n=10,x±s, bP<0.01, comparison between model group and normal control group and sham treatment group at the same time point; c P<0.05, d P < 0.01, compared with the model group at the same time point; e P<0.05, compared with the dexamethasone group at the same time point, f P<0.05, compared with the Lifei tablets group at the same time point.

[0316] Table 3. Effects of Baibei Yifei Capsule on the tension of isolated lung strips induced by endotoxin in rats with COPD

[0317]

[0318] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group; d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0319] Table 4. Effects of Baibei Yifei Capsule on the total and differential counts of leukocytes in bronchoalveolar lavage fluid of rats with COPD induced by endotoxin

[0320]

[0321]

[0322] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group,

[0323] f P<0.05, compared with the Lifei Tablets group.

[0324] Table 5. Effects of Baibei Yifei Capsule on antioxidant and inflammatory parameters of bronchoalveolar lavage fluid in rats with COPD induced by endotoxin

[0325]

[0326] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group,f P<0.05, compared with the Lifei Tablets group.

[0327] Table 6. Effects of Baibei Yifei Capsule on antioxidant and inflammatory indicators in lung tissue of rats with COPD induced by endotoxin

[0328]

[0329]

[0330] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0331] Table 7. Effects of Baibei Yifei Capsule on immune proteins in lung tissue of COPD rats induced by endotoxin

[0332]

[0333] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0334] Table 8. Effects of Baibei Yifei Capsule on lung function of smoke-induced COPD rats

[0335]

[0336] n=10,x±s, b P<0.01, comparison between model group and normal control group and sham treatment group at the same time point; c P<0.05, d P < 0.01, compared with the model group at the same time point; e P<0.05, compared with the dexamethasone group at the same time point, f P<0.05, compared with the Lifei tablets group at the same time point.

[0337] Table 9. Effects of Baibei Yifei Capsule on the tension of isolated lung strips in rats with COPD induced by smoke

[0338]

[0339] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0340] Table 10. Effects of Baibei Yifei Capsule on the total and differential counts of leukocytes in bronchoalveolar lavage fluid (BALF) of rats with COPD induced by smoke

[0341]

[0342] n=10,x±s, a P<0.05, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0343] Table 11. Effects of Baibei Yifei Capsule on antioxidant and inflammatory indicators in bronchoalveolar lavage fluid (BALF) of smoke-induced COPD rats

[0344]

[0345] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0346] Table 12. Effects of Baibei Yifei Capsule on antioxidant and inflammatory indexes in lung tissue of smoke-induced COPD rats

[0347]

[0348] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, fP<0.05, compared with the Lifei Tablets group.

[0349] Table 13. Effects of Baibei Yifei Capsule on immune indexes of lung tissue in smoke-induced COPD rats

[0350]

[0351] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0352] Table 14. Effects of Baibei Yifei Capsule on lung function in rats with bleomycin-induced pulmonary fibrosis

[0353]

[0354]

[0355] n=10,x±s, b P<0.01, model group compared with normal control group or sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0356] Table 15. Effect of Baibei Yifei Capsule on the tension of isolated lung strips in rats with bleomycin-induced pulmonary fibrosis

[0357]

[0358] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0359] Table 16. Effects of Baibei Yifei Capsule on the total and differential white blood cell counts in bronchoalveolar lavage fluid (BALF) of rats with bleomycin-induced pulmonary fibrosis

[0360]

[0361]

[0362] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0363] Table 17. Effects of Baibei Yifei Capsule on antioxidant and inflammatory indicators in bronchoalveolar lavage fluid (BALF) of rats with bleomycin-induced pulmonary fibrosis

[0364]

[0365] n=10,x±s, b P<0.01, model group compared with normal control group or sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0366] Table 18 Effects of Baibei Yifei Capsule on antioxidant and inflammatory indicators in lung tissue of rats with bleomycin-induced pulmonary fibrosis

[0367]

[0368]

[0369] n=10,x±s, b P<0.01, model group compared with normal control group or sham treatment group; c P<0.05, d P < 0.01, compared with the model group; e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0370] Table 19. Effects of Baibei Yifei Capsule on immune proteins in lung tissue of rats with bleomycin-induced pulmonary fibrosis

[0371]

[0372] n=10,x±s, b P<0.01, comparison between model group and normal control group; c P<0.05, d P < 0.01, compared with the model group;e P<0.05, compared with the dexamethasone group, f P<0.05, compared with the Lifei Tablets group.

[0373] Table 20. Effect of Baibei Yifei Capsule on xylene-induced ear swelling in mice

[0374]

[0375] n=10,x±s, a P<0.05, b P<0.01, compared with the blank control group; c P<0.05, d P<0.01, compared with aspirin and Lifei Tablets groups.

[0376] Table 21. Effects of Baibei Yifei Capsule on cotton ball granuloma in rats

[0377]

[0378] n=10,x±s, a P<0.05, b P<0.01, compared with the model control group; c P<0.05, compared with the dexamethasone group, d P<0.05, compared with the Lifei Tablets group.

[0379] Table 22. Minimum inhibitory concentration (MIC)

[0380]

[0381] Table 23. Effect of Baibei Yifei Capsule on cough induced by concentrated ammonia in mice

[0382]

[0383]

[0384] n=10,x±s, a P<0.05, b P<0.01, compared with the blank control group; c P<0.05, compared with the Lifei Tablets group; d P<0.01, compared with the codeine phosphate positive group.

[0385] Table 24. Effect of Baibei Yifei Capsule on citric acid-induced cough in guinea pigs

[0386]

[0387] n=10,x±s, cP<0.05, d P<0.01, compared with the blank control group; e P<0.05, compared with the codeine phosphate positive group, f P<0.05, compared with the Lifei Tablets group.

[0388] Table 25. Effect of Baibei Yifei Capsule on cough induced by electrical stimulation of superior laryngeal nerve in cats

[0389]

[0390] n=10,x±s, a P<0.05, b P<0.01, compared with the same group before administration; c P<0.05, d P<0.01, compared with the blank control group; e P<0.05, compared with the codeine group, f P<0.05, compared with the Chinese medicine positive control group.

[0391] Table 26. Effect of Baibei Yifei Capsule on nebulization-induced wheezing in guinea pigs

[0392]

[0393] n=10,x±s, c P<0.05, d P<0.01, compared with the blank control group; e P<0.05, compared with the aminophylline group, f P<0.05, compared with the Lifei Tablets group.

[0394] Table 27. Effects of Baibei Yifei Capsule on histamine-induced changes in tension of isolated guinea pig tracheal smooth muscle

[0395]

[0396] n=6,x±s, b P < 0.01, compared with the saline group; c P<0.01, compared with the aminophylline group.

[0397] Table 28. Effect of Baibei Yifei Capsule on expectoration in mice (tracheal segment phenol red method)

[0398]

[0399] n=10,x±s, d P<0.01, compared with the blank control group; e P<0.05, compared with the ammonium chloride group, fP<0.05, compared with the Lifei Tablets group.

[0400] Table 29. Effect of Baibei Yifei Capsule on expectoration in rats (capillary tube method)

[0401]

[0402] n=10,x±s, c P<0.05, d P<0.01, compared with the blank control group; e P<0.05, compared with the ammonium chloride group, f P<0.05, compared with the Lifei Tablets group.

[0403] 3. Clinical trials of Baibei Yifei capsules in the treatment of COPD

[0404] Objective: To further evaluate the efficacy and safety of Baibei Yifei Capsule in the treatment of chronic obstructive pulmonary disease (COPD) with lung-yin deficiency, based on its confirmed clinical indications.

[0405] Methods: According to the inclusion criteria, 120 patients with COPD (lung yin deficiency syndrome) were selected. The patients were randomly divided into the experimental group and the control group, with 90 cases in the experimental group and 30 cases in the control group. The experimental group: took Baibei Yifei capsules orally, 3 capsules each time, 3 times a day. The control group: took Yangyin Qingfei pills orally, 1 pill each time, 3 times a day. The course of medication was 2 weeks. The changes of TCM syndromes and single main symptoms, secondary symptoms and laboratory test indicators (blood routine, liver and kidney function, urine routine, stool routine and electrocardiogram) of the patients in the experimental group and the control group before and after treatment were observed, and the effectiveness and safety of the experimental and control drugs were evaluated.

[0406] result:

[0407] 1. The total score of TCM syndrome and the scores of main and secondary TCM symptoms before and after treatment were compared between the experimental group and the control group. The results showed that the differences were significant (P < 0.01), indicating that both groups of drugs had obvious therapeutic effects.

[0408] 2. In terms of improving the efficacy of TCM syndrome, the effective rate of the experimental group after treatment was 83.75%, and the effective rate of the control group was 75.0%. After the rank sum test, the difference between the experimental group and the control group was statistically significant (P < 0.05), indicating that the experimental group was superior to the control group in improving the efficacy of TCM syndrome.

[0409] 3. Comparison of changes in the scores of individual TCM symptoms between the two groups of patients before and after treatment: the experimental group was significantly better than the control group in improving cough and asthma symptoms (P < 0.01); it was better than the control group in improving chest tightness and shortness of breath symptoms (P < 0.05), while there was no significant difference in improving sputum color and dry mouth and throat symptoms between the two groups (P > 0.05).

[0410] 4. Comparison of lung function before and after treatment: There was no statistically significant difference in the lung function of the two groups before and after treatment (P>0.05), indicating that the experimental group and the control group had no obvious effect in improving lung function. However, according to the table, the lung function of the patients in the experimental group improved after treatment, while the lung function of the patients in the control group improved little.

[0411] 5. There was no statistically significant difference in blood routine, liver function and kidney function between the two groups before and after treatment (P>0.05). No obvious abnormal changes were found in other safety indicators, and no obvious adverse reactions and side effects occurred during the experiment.

[0412] Conclusion: Baibei Yifei Capsule has the effect of improving the traditional Chinese medicine symptoms in the treatment of chronic obstructive pulmonary disease (COPD) with lung yin deficiency, and is better than the control group in improving symptoms such as cough, wheezing, chest tightness and shortness of breath. It is safe to use, and no obvious adverse reactions and toxic side effects were found.

[0413] 4. Clinical trials of Baibei Yifei capsules in the treatment of pulmonary tuberculosis

[0414] 1 Clinical Data

[0415] 1.1 Case source

[0416] The 72 patients observed and studied in this trial were all inpatients and outpatients from Yunnan Provincial Infectious Diseases Hospital. They met the inclusion criteria and were divided into a treatment group and a control group according to a random number table, with 36 cases in each group, taking into account the dropout factors.

[0417] 1.2 Diagnostic criteria

[0418] 1.2.1 Western medicine diagnostic criteria

[0419] It complies with the diagnostic criteria for pulmonary tuberculosis in the "Health Industry Standard of the People's Republic of China - Diagnosis of Tuberculosis (WS288-2017)" issued on November 9, 2017, and the "WHO Guidelines for the Treatment and Care of Drug-Sensitive Tuberculosis (2017 Update)", which is the latest revision of the "Guidelines for the Diagnosis and Treatment of Tuberculosis (Fourth Edition)" formulated by the WHO in 2009. On January 13, 2016, the National Institute for Health and Care Excellence of the United Kingdom issued the 2016 NICE Guidelines: Tuberculosis (NG.33).

[0420] 1.2.2 Standards for TCM Syndrome Differentiation

[0421] The diagnostic criteria of TCM syndromes are based on the Guidelines for the Diagnosis and Treatment of Common Diseases in TCM Internal Medicine (Part on TCM Syndromes) (issued by the China Association of Traditional Chinese Medicine on August 5, 2008) and the diagnostic basis, syndrome classification, and efficacy evaluation of pulmonary tuberculosis - the TCM industry standard of the People's Republic of China "Standards for the Diagnosis and Efficacy of TCM Internal Medicine Diseases" (ZY / T001.1-94) and "Chinese Internal Medicine" (Edited by Wu Mianhua and Wang Xinyue in 2012).

[0422] Lung Yin Deficiency Syndrome: (1) Dry cough without sputum, short cough, or coughing up a small amount of sticky sputum, or sputum with blood; (2) Hot flashes and night sweats, accompanied by dull pain in the chest, dry mouth and throat, hot hands and soles, and dry skin; (3) Red tongue, thin tongue coating, and thin and rapid pulse.

[0423] If two of the above items (1) are present, combined with items (2) and (3), the syndrome can be diagnosed as lung yin deficiency.

[0424] 1.3 Efficacy determination

[0425] 1.3.1 Syndrome grading quantitative table, see Table 30.

[0426] Table 30 Quantitative table for the classification of lung yin deficiency syndrome in pulmonary tuberculosis

[0427]

[0428] Note: Mild, 0-7 points, moderate, 8-14 points, severe, 15-21 points.

[0429] 1.3.2 Sputum smear results, see Table 31.

[0430] Table 31 Sputum smear results evaluation criteria

[0431] Classification Acid-fast bacilli evaluate 1 No acid-fast bacilli were found in 300 consecutive observations of different fields of view - 2 Reports the number of acid-fast bacilli, 1-8 / 300 fields of view 3 Report acid-fast bacilli positive, 3-9 / 100 fields of view + 4 Report acid-fast bacilli positive, 1-9 / 10 fields of view ++ 5 Report acid-fast bacilli positive, 1-9 / per field of view +++ 6 Report acid-fast bacilli positive, ≥10 / per field of view ++++

[0432] 1.3.3 Sputum culture results, see Table 32.

[0433] Table 32 Evaluation criteria for sputum culture results

[0434] Classification Mycobacterium tuberculosis evaluate 1 No colony growth reported, culture negative - 2 When the colony growth is less than 1 / 4 of the slope area Actual colony count 3 The colony occupies 1 / 4 of the slope area + 4 The colony occupies 1 / 2 of the slope area ++ 5 The colony occupies 3 / 4 of the slope area +++ 6 The colonies cover the entire surface of the culture medium ++++

[0435] Note: For statistical convenience, only the sputum smear or sputum culture results of patients in the treatment group and the control group were divided into negative and positive.

[0436] 1.3.4 Disease severity classification standards

[0437] (1) Obvious absorption: the lesion absorbs ≥1 / 2 of the original lesion;

[0438] (2) Absorption: lesion absorption <1 / 2 of the original lesion;

[0439] (3) No change: no obvious changes in the lesion;

[0440] (4) Worsening: the lesion expands or spreads.

[0441] 1.4 Test case criteria

[0442] 1.4.1 Criteria for inclusion of cases

[0443] (1) Patients who meet the Western medical diagnostic criteria for pulmonary tuberculosis;

[0444] (2) Meet the diagnostic criteria of TCM for pulmonary tuberculosis and lung yin deficiency syndrome;

[0445] (3) Aged between 18 and 70 years old;

[0446] (4) Sign the informed consent and voluntarily participate in the experiment.

[0447] Anyone who meets the above criteria can be included in the trial.

[0448] 1.4.2 Exclusion criteria

[0449] (1) Those who do not meet any of the inclusion criteria;

[0450] (2) Patients who refuse Chinese medicine treatment;

[0451] (3) Patients with allergic constitution or a history of allergy to multiple drugs, or those who are severely allergic to any drug in this trial;

[0452] (4) Patients with liver, kidney, autoimmune diseases, and severe lung or skin infections;

[0453] (5) Patients whose condition is critical and it is difficult to make an accurate evaluation of the effectiveness and safety of the drug;

[0454] (6) Incomplete data may affect the judgment of efficacy;

[0455] (7) Those who meet the diagnostic criteria of traditional Chinese and Western medicine but are currently participating in other clinical trials.

[0456] Any test that meets the above criteria can be excluded.

[0457] 1.4.3 Dropout criteria

[0458] (1) Allergic reactions or severe adverse reactions occur, and the trial should be stopped according to the doctor's judgment;

[0459] (2) poor compliance of subjects;

[0460] (3) During the trial, patients develop secondary infection, severe complications, and special physiological changes;

[0461] (4) Patients who voluntarily request to withdraw from the trial to their attending physician;

[0462] (5) Patients who have not withdrawn from the trial but no longer receive treatment, medication or examinations and have lost contact.

[0463] For all dropout cases, the reasons for dropout should be filled in the case report form. If the patient has adverse reactions, the adverse reaction statistics form should be filled in.

[0464] 1.4.4 Criteria for exclusion of cases

[0465] (1) Those who were mistakenly included and did not meet the inclusion criteria;

[0466] (2) Those who met the inclusion criteria but never took medication after inclusion, or did not take medication according to the trial regulations, or had no follow-up records;

[0467] (3) Those found to be infected with non-tuberculosis bacteria after bacterial typing.

[0468] 1.4.5 Test termination criteria

[0469] (1) Problems that seriously endanger patient safety occur during the trial;

[0470] (2) The trial found that the drug treatment effect was too poor and had low clinical value;

[0471] (3) Major errors in the experimental design or important deviations in the experiment were found during the trial, making it difficult to evaluate the drug effect;

[0472] (4) The researcher requests termination;

[0473] (5) The competent authority requires the cancellation of the test, etc.

[0474] 2 Test methods

[0475] 2.1 Experimental design

[0476] (1) Design method: A stratified randomization, block randomization, and parallel controlled trial design method was adopted. All clinical trials were completed by the trialists in the inpatient department and outpatient clinic of Yunnan Provincial Infectious Disease Hospital.

[0477] (2) Sample size: The estimated number of test cases is 60, with a 1:1 ratio between the treatment group and the control group. Taking into account the dropout factor, the number of cases is increased by 20%, that is, 6 cases are added to each group. Therefore, the total number of test cases is 72, with 36 cases in the treatment group and 36 cases in the control group.

[0478] (3) Random grouping method: A random number table was used to randomly assign 72 subjects (36 in the treatment group and 36 in the control group).

[0479] (4) Control method: The control group was given routine anti-tuberculosis treatment in Western medicine. The treatment group was given oral Baibei Yifei capsules on this basis. The efficacy and toxicity reduction were compared to provide a reference for the combination of Yi medicine and conventional Western medicine for clinical tuberculosis treatment.

[0480] (5) Blind design: Since the treatment methods of the treatment group and the control group were different, this trial adopted an “open” method, that is, no blind design was performed.

[0481] 2.2 Treatment options

[0482] 2.2.1 Name and specification of the investigational drug

[0483] Baibei Yifei Capsule, a Yi medicine produced by Yunnan Yongzitang Pharmaceutical Co., Ltd. Specification: 0.3g*12s*2 plates, approval number: National Medicine Standard Z20025124.

[0484] 2.2.2 Medication regimen

[0485] Both the treatment group and the control group received routine basic medical treatment.

[0486] (1) Treatment group: On the basis of routine basic treatment in internal medicine, Baibei Yifei Capsule was used, 3 capsules each time, 3 times a day.

[0487] ⑵Control group: Routine basic medical treatment. According to the Ministry of Health's "Outpatient Diagnosis and Treatment Guidelines for Pulmonary Tuberculosis" (2012 edition) [8] , the recommended drug treatment regimen is implemented:

[0488] ① Initial treatment of pulmonary tuberculosis: 2HRZE / 4HR, 2H3R3Z3E3 / 4H3R3.

[0489] ② Re-treatment of pulmonary tuberculosis: 2HRZES / 6HRE, 3HRZE / 6HRE. Patients with drug sensitivity test results can develop a treatment plan based on the results of the drug sensitivity test and previous medication history.

[0490] H: isoniazid, R: rifampicin, Z: pyrazinamide, E: ethambutol, S: streptomycin.

[0491] While undergoing chemotherapy, the patient's complications or comorbidities can be treated.

[0492] ⑶Treatment course: 4 weeks.

[0493] 2.2.3 Combined medication

[0494] (1) Except for the drugs specified by the trial institute, it is prohibited to take other Chinese or Western medicines that may affect the efficacy of the trial drugs;

[0495] (2) Any medication or treatment required for the concurrent disease must be recorded in detail in the case report form (including the name of the medication, dosage and time of use, the name of the therapy, number of times used and time of use, etc.).

[0496] 2.3 Preparation and Administration of Trial Drugs

[0497] (1) Dispense the medicines according to the corresponding numbers of the drugs to the corresponding patients one by one;

[0498] (2) During each follow-up visit, the researcher must truthfully record the amount of medication used by the subject and the time of use, and record it carefully and promptly.

[0499] 2.4 Observation indicators and observation time points

[0500] 2.4.1 Safety indicators

[0501] (1) Routine blood, urine and stool tests;

[0502] (2) liver function (ALT, AST), renal function (UA, BUN, Cr);

[0503] (3) Chest CT.

[0504] The above examination indicators should be performed once before and after treatment. If the examination results are normal before treatment but abnormal after treatment, re-examination should be performed until they return to normal.

[0505] 2.4.2 Efficacy indicators

[0506] (1) Evaluation of TCM syndrome efficacy;

[0507] (2) Disease efficacy evaluation indicators.

[0508] 2.4.3 Observation time

[0509] (1) After the start of the trial, the subjects will return for a follow-up visit once after the second week and once after the fourth week, and record their symptoms and signs, as well as adverse events, concomitant medications, etc.

[0510] (2) After the trial medication is completed, the TCM syndrome scores, sputum bacteria examination results, and chest imaging lesion absorption status will be reviewed, and a comparative evaluation of the efficacy and cure rate will be performed.

[0511] (3) Follow-up period: After the end of the trial drug treatment, the subjects must conduct a telephone follow-up (2 months).

[0512] 2.5 Criteria for determining efficacy

[0513] 2.5.1 Evaluation of TCM Syndrome Efficacy

[0514] (1) Clinical recovery: TCM clinical symptoms and signs disappear or almost disappear, and syndrome score decreases by ≥95%;

[0515] (2) Significantly effective: clinical symptoms and signs of TCM were significantly improved, and the syndrome score was reduced by ≥70%;

[0516] (3) Effective: clinical symptoms and signs of TCM have improved, and the syndrome score has decreased by ≥30%;

[0517] (4) Ineffective: There was no significant improvement in TCM clinical symptoms and signs or they even got worse, and the syndrome score decreased by less than 30%.

[0518] Note: Calculation formula: Efficacy index = (sum of scores before treatment - sum of scores after treatment) / sum of scores before treatment × 100%

[0519] 2.5.2 Disease efficacy evaluation

[0520] (1) Before treatment, the experimenter collected one sample of the subject's instant sputum, morning sputum, and night sputum for smear examination or sputum culture examination. At the end of the fourth week of treatment, another sample of random sputum, morning sputum, and night sputum was collected for smear examination or sputum culture examination.

[0521] Sputum negative conversion rate = (number of patients with sputum negative conversion / number of patients registered with smear-positive or sputum culture-positive results) × 100%.

[0522] (2) Chest CT scans were performed once before treatment and once every 4 weeks after treatment to compare the absorption of lesions in chest images before and after treatment. The evaluation method was divided into the following according to the standards established by the National Tuberculosis Society in 1982:

[0523] Chest CT lesion efficacy evaluation:

[0524] Obvious absorption: the lesion absorbs ≥1 / 2 of the original lesion;

[0525] Absorption: lesion absorption <1 / 2 of the original lesion;

[0526] No change: no obvious changes in the lesion;

[0527] Worsening: The lesion enlarges or spreads.

[0528] 2.6 Statistical analysis methods

[0529] SPSS 22.0 software was used for statistical analysis. The measurement data of each group that met the normal distribution were statistically described using mean ± standard deviation, the paired sample t test was used for comparison within the same group, the two independent sample t test was used for comparison between groups, and the count data were analyzed using X-ray diffraction analysis. 2 For the ranked data and the measurement data that did not conform to the normal distribution, the rank sum test was used. Both the t test and the rank sum test were considered statistically significant when P < 0.05.

[0530] 3 Data and statistical results

[0531] 3.1 General data analysis

[0532] The 72 patients with pulmonary tuberculosis studied were all inpatients and outpatients of Yunnan Provincial Infectious Diseases Hospital from December 2017 to March 2019. Among them, 4 subjects withdrew from the trial and 4 subjects were lost because they took other therapeutic drugs by themselves, which affected the efficacy and safety of this trial. After the dropout, 64 cases remained, including 31 cases in the treatment group and 33 cases in the control group.

[0533] 3.1.1 Gender

[0534] Among the 64 cases that met the requirements of the study, there were 17 males and 14 females in the treatment group and 17 males and 16 females in the control group. The gender composition of the treatment group and the control group is shown in Table 33. Figure 1 .

[0535] Table 33 Comparison of gender composition between the two groups

[0536]

[0537] The gender of the subjects in the treatment group and the control group were compared. 2 The test results were not statistically significant (P>0.05) and were comparable.

[0538] 3.1.2 Age

[0539] Among the 64 patients who met the requirements of the study, 31 patients in the treatment group were aged between 18 and 70 years old, with an average age of 44.19±14.54 years old; 33 patients in the control group were aged between 18 and 70 years old, with an average age of 44.18±15.42 years old. The age group was tested to be in line with normal distribution, and the independent sample t test was used for testing, t=-0.003, p=0.998>0.05, indicating that there was no statistically significant difference in the age distribution of the two groups of patients, suggesting that the two groups of patients were comparable in age. See Table 34.

[0540] Table 34 Comparison of age groups between the treatment group and the control group

[0541]

[0542] Table 34: The independent sample t-test was used to compare the ages of the subjects in the treatment group and the control group of this experiment, and it was found that there was no statistical significance (P>0.05).

[0543] 3.1.3 Course of disease

[0544] Among the 64 patients who met the study, since the course of disease in the two groups did not conform to normal distribution, the course of disease was expressed as median ± interquartile range, including 31 cases in the treatment group, expressed as: 23 ± 70.5; 33 cases in the control group, expressed as: 6 ± 101. See Table 35.

[0545] Table 35 Comparison of disease course between the two groups of patients

[0546]

[0547] Table 35: Since the course of disease in the treatment group and the control group of this trial did not conform to the normal distribution, we expressed the course of disease as median ± interquartile range. The course of disease in the treatment group and the control group was compared by rank sum test and found to be insignificant (P>0.05).

[0548] 3.1.4 Comparison of TCM syndrome scores between groups before treatment

[0549] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, and the score before treatment was 7.65±1.76; 33 were in the control group, and the score before treatment was 7.64±2.23. t: 0.017, P: 0.986. See Table 36.

[0550] Table 36 Comparison of TCM syndrome scores between the two groups before treatment

[0551]

[0552] Table 36: The TCM syndrome scores of the treatment group and the control group before treatment were compared using an independent sample t-test, and the results were not statistically significant (P>0.05).

[0553] 3.1.5 Comparison of sputum smear or sputum culture between groups before treatment

[0554] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, and the mean rank of sputum smear or sputum culture before treatment was 32.65, and the rank sum was 1012.00; 33 were in the control group, and the mean rank of sputum smear or sputum culture before treatment was 32.36, and the rank sum was 1068.00. Z: -0.073, P: 0.942. See Table 37, Fig.13 .

[0555] Table 37 Comparison of sputum smear or sputum culture groups before treatment

[0556]

[0557] Table 37: The sputum smear or sputum culture results (divided into negative and positive) of patients in the treatment group and the control group of this trial before treatment were ranked data. After the rank sum test, the sputum smear or sputum culture results of patients in the treatment group and the control group before treatment were compared, and it was found that there was no statistical significance (P>0.05).

[0558] 3.2 Comparison of efficacy

[0559] 3.2.1 Intra-group comparison of sputum smear or sputum culture results before and after treatment

[0560] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, and the mean rank of sputum smear or sputum culture after treatment was 8.50, the rank sum was 136.00, Z: -4.000, P: 0.000; and 33 were in the control group, and the mean rank of sputum smear or sputum culture after treatment was 5.00, the rank sum was 45.00, Z: -3.000, P: 0.003. See Table 38.

[0561] Table 38 Comparison of sputum smear or sputum culture within the group before and after treatment

[0562] Group Number of cases Rank Mean Rank Sum Z P Treatment Group 31 8.50 136.00 -4.000 0.000 Control group 33 5.00 45.00 -3.000 0.003

[0563] Table 38: After the treatment, the paired rank sum test was used to compare the sputum smear or sputum culture results before and after treatment in the treatment group and the control group, and the results were statistically significant (P<0.01). This shows that both treatment methods in this trial have good therapeutic effects.

[0564] 3.2.2 Comparison of TCM syndrome scores within each group before and after treatment

[0565] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, and the TCM syndrome score before treatment was 7.65±1.76, and the score after treatment was 2.81±1.89. t: 14.742, P: 0.000; 33 were in the control group, and the TCM syndrome score before treatment was 7.64±2.23, and the score after treatment was 4.00±2.36. t: 11.608, P: 0.000. See Table 39.

[0566] Table 39 Comparison of TCM syndrome scores within the same group before and after treatment

[0567] Group n Pre-treatment score After treatment score t P Treatment Group 31 7.65±1.76 2.81±1.89 14.742 0.000 Control group 33 7.64±2.23 4.00±2.36 11.608 0.000

[0568] Table 39: Paired sample t-test showed that after the treatment, the TCM syndrome scores of the treatment group and the control group were compared before and after the treatment, and the results were statistically significant (P<0.01). This shows that from the perspective of TCM syndrome, both treatment methods in this trial have good therapeutic effects.

[0569] 3.2.3 Comparison of sputum smear or sputum culture results between groups after treatment

[0570] Among the 64 patients who met the requirements of the study, 31 were in the treatment group. The mean rank of sputum smear or sputum culture after treatment was 28.63, the rank sum was 887.50, 20 patients were positive before treatment, 16 patients turned negative, and the negative conversion rate was 80%; 33 were in the control group. The mean rank of sputum smear or sputum culture after treatment was 36.14, the rank sum was 1192.50, 21 patients were positive before treatment, 9 patients turned negative, and the negative conversion rate was 42.86%. Z: -2.149, P: 0.032. See Table 40, Table 41, Fig.14 .

[0571] Table 40 Comparison of sputum smear or sputum culture results between the two groups after treatment

[0572]

[0573] Table 41 Negative conversion rate of smear-positive patients

[0574] Grouping Positive patients Patients with negative Negative conversion rate (%) Treatment Group 20 16 80.00 Control group 21 9 42.86

[0575] Table 40, Table 41: After the treatment, the rank sum test was used to compare the sputum smear or sputum culture results between the treatment group and the control group, and the results were statistically significant (P < 0.05). The number of patients with positive sputum smear or sputum culture after treatment in the treatment group was lower than that in the control group, indicating that the treatment effect of the treatment group was better than that of the control group.

[0576] 3.2.4 Comparison of TCM syndrome scores between groups after treatment

[0577] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, and the score after treatment was 2.81±1.89; 33 were in the control group, and the score after treatment was 4.00±2.36. t: -2.226, P: 0.030. See Table 42.

[0578] Table 42 Comparison of TCM syndrome groups after treatment

[0579]

[0580] Table 42: The TCM syndrome scores of the treatment group and the control group after treatment were compared by independent sample t-test, and the results showed statistically significant differences (P value < 0.05). The TCM syndrome scores of the treatment group were lower than those of the control group after treatment. From the perspective of TCM syndrome, the therapeutic effect of the treatment group in this trial was better than that of the control group.

[0581] 3.2.5 Comparison of efficacy after treatment

[0582] Among the 64 patients who met the requirements of the study, 31 were in the treatment group. After treatment, there were no obvious absorption cases of chest CT lesions, 27 were absorbed, 4 were unchanged, and no deterioration cases were found. The total absorption rate was 87.10%. Among the 33 patients in the control group, there were no obvious absorption cases of chest CT lesions, 21 were absorbed, 12 were unchanged, and no deterioration cases were found. Z: -2.149; P: 0.032. Among the 31 patients in the treatment group, 3 were cured, 8 were markedly effective, 19 were effective, and 1 was ineffective according to the TCM symptoms after treatment. The total effective rate was 96.77%. Among the 33 patients in the control group, 1 was cured, 7 were markedly effective, 15 were effective, and 10 were ineffective according to the TCM symptoms after treatment. Z: -2.231; P: 0.026. See Table 43, Table 44, Fig.15 , Fig.16 .

[0583] Table 43 Chest CT efficacy evaluation

[0584]

[0585]

[0586] Table 43: The rank sum test of the efficacy of the treatment group and the control group of this experiment was statistically significant (P < 0.05). The absorption rate of the lesion in chest imaging of the treatment group was higher than that of the control group.

[0587] Table 44 Comparison of efficacy of TCM syndromes

[0588]

[0589] Table 44: The rank sum test of the efficacy of TCM syndromes in the treatment group and the control group of this experiment was performed, and the result was statistically significant (P<0.05), indicating that the effective rate of treatment based on TCM syndromes in the treatment group was higher than that in the control group.

[0590] 3.3 Safety indicators

[0591] 3.3.1 Comparison of blood routine and renal function in each group before and after treatment

[0592] Among the 64 patients who met the requirements of the study, 31 were in the treatment group and 33 were in the control group (see Table 45).

[0593] Table 45 Comparison of blood routine and renal function in each group before and after treatment

[0594]

[0595] Table 45: The paired sample t test showed that the RBC value before and after the treatment group was statistically significant (P<0.05), and the RBC value after treatment was lower than before and within the normal range. Except for the above items, there was no statistical significance in the comparison of other indicators before and after treatment (P>0.05).

[0596] 3.3.2 Comparison of serum uric acid and liver function between the two groups before treatment

[0597] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, with UA: 294.10±42.13, ALT: 19.33±7.02, AST: 21.77±6.87 before treatment. There were 33 patients in the control group, with UA: 295.94±41.33, ALT: 19.70±8.40, AST: 21.21±6.26 before treatment. See Table 46.

[0598] Table 46 Comparison of UA, ALT and AST between groups before treatment

[0599] Group Number of cases UA ALT AST Treatment Group 31 294.10±42.13 19.33±7.02 21.77±6.87 Control group 33 295.94±41.33 19.70±8.40 21.21±6.26

[0600] There was no statistically significant difference in UA between the treatment group and the control group before treatment, t=-0.177, P=0.860, P>0.05.

[0601] There was no statistically significant difference in ALT between the treatment group and the control group before treatment, t=-0.193, P=0.848, P>0.05.

[0602] There was no statistically significant difference in AST between the treatment group and the control group before treatment (t=0.342, P=0.733, P>0.05).

[0603] 3.3.3 Comparison of blood uric acid indexes before and after treatment

[0604] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, with a score of 294.10±42.13 before treatment and 504.58±83.69 after treatment, t: -17.272, P: 0.000. There were 33 patients in the control group, with a score of 295.94±41.33 before treatment and 555.67±89.77 after treatment, t: -14.601, P: 0.000. See Table 47.

[0605] Table 47 Comparison of serum uric acid in each group before and after treatment

[0606] Group Number of cases Before treatment After treatment t P Treatment Group 31 294.10±42.13 504.58±83.69 -17.272 0.000 Control group 33 295.94±41.33 555.67±89.77 -14.601 0.000

[0607] After 4 weeks of treatment: comparison of UA between the two groups, t=2.351, P=0.022, P<0.05.

[0608] After treatment, the blood uric acid levels of both the treatment group and the control group increased (P < 0.05), indicating that anti-tuberculosis drugs have a side effect of increasing the blood uric acid level of the subjects, but the degree of increase in the blood uric acid level of the subjects in the treatment group was lower than that in the control group (P < 0.05), indicating that Baibei Yifei Capsule has the effect of lowering blood uric acid during the treatment course.

[0609] 3.3.4 Comparison of gastrointestinal symptoms and allergic reactions in patients before treatment

[0610] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, 17 of whom had gastrointestinal symptoms and 13 of whom had mild allergic reactions before treatment, and 33 were in the control group, 18 of whom had gastrointestinal symptoms and 16 of whom had mild allergic reactions before treatment. See Table 48.

[0611] Table 48 Comparison of gastrointestinal symptoms and mild allergic reactions before treatment Comparison between groups

[0612]

[0613] Note: Gastrointestinal symptoms include: subjects experiencing nausea and vomiting, abdominal distension and pain, loss of appetite and other side effects; mild allergic reactions include: subjects experiencing mild rash, itching, etc.

[0614] There was no statistical difference in the number of gastrointestinal symptoms and mild allergic reactions between the two groups before treatment (P>0.05), indicating that there was no significant relationship between the number of gastrointestinal symptoms and mild allergic reactions between the two groups before treatment.

[0615] 3.3.5 Comparison of the number of cases of abnormal liver function, gastrointestinal symptoms and allergic reactions after treatment

[0616] Among the 64 patients who met the requirements of the study, 31 were in the treatment group, 5 of whom had abnormal ALT, 8 of whom had abnormal AST, 15 of whom had gastrointestinal symptoms, and 10 of whom had mild allergic reactions after treatment. There were 33 patients in the control group, 14 of whom had abnormal ALT, 17 of whom had abnormal AST, 26 of whom had gastrointestinal symptoms, and 12 of whom had mild allergic reactions after treatment. See Table 49.

[0617] Table 49 Comparison of abnormal ALT, AST, gastrointestinal symptoms and mild allergic reactions between groups after treatment

[0618]

[0619] Note: Gastrointestinal symptoms include: subjects experiencing nausea and vomiting, abdominal distension and pain, loss of appetite and other side effects; mild allergic reactions include: subjects experiencing mild rash, itching, etc.

[0620] There were statistically significant differences in ALT and AST abnormalities between the two groups of patients after treatment (P < 0.05), indicating that Baibei Yifei Capsules had a more obvious liver-protecting and enzyme-lowering effect during the treatment; there were statistically significant differences in the number of gastrointestinal symptoms between the two groups of patients after treatment (P < 0.05), indicating that Baibei Yifei Capsules were effective in alleviating patients' gastrointestinal symptoms during the treatment; while there was no statistically significant difference in the number of mild allergic reaction symptoms after treatment (P > 0.05), indicating that Baibei Yifei Capsules had no obvious effect on alleviating patients' mild allergic reactions during the treatment.

[0621] 3.3.6 Routine status of bowel movements

[0622] During the trial, no obvious abnormalities were found in the stool and urine routine of patients in the treatment group and the control group before and after treatment, indicating that the two treatment regimens in this trial had no obvious effect on the stool and urine routine.

[0623] 4 Clinical efficacy analysis

[0624] This design adopts a randomized, controlled clinical trial design method, and randomly divides 72 patients with pulmonary tuberculosis who meet the TCM syndrome differentiation into 36 control groups and 36 treatment groups. The 72 patients with pulmonary tuberculosis studied were all inpatients and outpatients of Yunnan Provincial Infectious Disease Specialist Hospital from December 2017 to March 2019. Four subjects withdrew from the trial due to self-administration of other therapeutic drugs that affected efficacy and safety, and 4 subjects lost contact. Among the remaining 64, 31 were in the treatment group and 33 were in the control group. Both groups received routine basic treatment. Because Western medicine anti-tuberculosis drugs are still the conventional treatment for pulmonary tuberculosis, this trial selected routine basic treatment as the basic control, and the treatment group was treated with Baibei Yifei Capsules on this basis to compare its superiority.

[0625] 4.1 Evaluation of the efficacy of chest imaging lesion absorption

[0626] The total absorption rate of the treatment group was 87.10%; the total effective rate of the control group was 63.64%. The comparison of chest imaging efficacy between the two groups was statistically significant (P < 0.05). This shows that the lesion absorption rate of the treatment group was greater than that of the control group, and both treatments had therapeutic effects, and the treatment group was better than the control group.

[0627] 4.2 Evaluation of sputum smear or sputum culture results

[0628] The sputum smear or sputum culture results of the two groups of patients before and after treatment were compared within the group, and the results were statistically significant (P value < 0.01), indicating that both treatment regimens had therapeutic effects.

[0629] The sputum smear or sputum culture results of the treatment group and the control group before treatment were compared, and the results were not statistically significant (P value>0.05), indicating that there was no significant difference in the sputum smear or sputum culture results of the two groups of patients before treatment.

[0630] The sputum smear or sputum culture results of the two groups of patients after treatment were compared, and there was a statistical difference (P<0.05). The sputum smear or sputum culture results of the treatment group were lower than those of the control group after treatment. The negative conversion rate of sputum bacteria: the negative conversion rate of the treatment group was 80.00%; the negative conversion rate of the control group was 42.86%. This shows that the treatment effect of the treatment group is better than that of the control group.

[0631] 4.3 Evaluation of TCM Syndrome Accumulation

[0632] The total effective rate of the treatment group was 96.77%, and the cure rate was 9.68%; the total effective rate of the control group was 69.70%, and the cure rate was 3.03%. The efficacy of the treatment group was better than that of the control group, with statistical significance (P<0.05).

[0633] The TCM syndrome scores of the treatment group and the control group after treatment were compared within the group, and the results showed significant statistical differences (P < 0.01), indicating that both treatment plans can improve TCM syndrome symptoms.

[0634] The TCM syndrome scores of the treatment group and the control group before treatment were compared between the two groups, and the results were not statistically significant (P>0.05), indicating that there was no difference before treatment. The TCM syndrome scores of the treatment group and the control group after treatment were compared between the two groups, and the results were statistically significant (P<0.05). And by comparing the TCM syndrome scores of the two groups of patients, it was found that the scores of the patients in the treatment group after treatment were lower than those in the control group, which shows that the treatment plan of the treatment group is more advantageous than that of the control group.

[0635] 4.4 Safety evaluation

[0636] The UA of the patients in the treatment group and the control group increased (P < 0.05), and the degree of increase of UA in the treatment group after the treatment was lower than that in the control group (P < 0.05), indicating that anti-tuberculosis Western medicine has the side effect of increasing the patient's blood uric acid, and Baibei Yifei Capsule can reduce the patient's blood uric acid during the treatment process; the abnormalities of ALT and AST in the two groups of patients were statistically different after treatment (P < 0.05), indicating that Baibei Yifei Capsule has a more obvious effect of protecting the liver and reducing enzymes during the treatment course; the number of gastrointestinal symptoms in the two groups of patients was statistically different after treatment (P < 0.05), indicating that Baibei Yifei Capsule is effective in alleviating the gastrointestinal symptoms of patients during the treatment course; and the number of mild allergic reaction symptoms was not statistically significant after treatment (P > 0.05), indicating that Baibei Yifei Capsule has no obvious effect on alleviating mild allergic reactions in patients during the treatment course. The other safety indicators were all within the normal range.

[0637] The effect of Baibei Yifei Capsule combined with conventional basic treatment is better than that of conventional basic treatment alone. It is safe to use and is effective in reducing the toxic and side effects of conventional Western medicine treatment.

[0638] 5 Conclusion

[0639] 5.1 Clinical observation of Baibei Yifei Capsule combined with routine anti-tuberculosis basic treatment of pulmonary tuberculosis showed that most patients with pulmonary tuberculosis treated with Baibei Yifei Capsule combined with routine anti-tuberculosis basic treatment had significant improvement in symptoms and signs within one course of treatment (4 weeks). The chest CT lesion absorption rate in the treatment group was greater than that in the control group, and the sputum bacteria negative conversion rate was higher than that in the control group. This shows that Baibei Yifei Capsule combined with routine anti-tuberculosis basic treatment of internal medicine can promote lesion absorption and accelerate the speed of sputum bacteria negative conversion, and the treatment effect is significantly better than that of simple routine anti-tuberculosis basic treatment of internal medicine.

[0640] 5.2 During the entire treatment process, no subjects experienced worsening of their condition due to taking Baibei Yifei Capsules. Baibei Yifei Capsules have the effect of reducing patients' blood uric acid; have a more obvious liver protection and enzyme reduction effect; are effective in alleviating patients' gastrointestinal symptoms; but have no obvious effect on alleviating patients' mild allergic reactions. The remaining safety indicators are all within the normal range. Baibei Yifei Capsules are safe to use and have a significant effect in reducing the toxic and side effects of conventional Western medicine treatments.

[0641] In summary, Baibei Yifei Capsule combined with conventional Western medicine anti-tuberculosis basic treatment is better than conventional Western medicine basic treatment alone. It is safe to use and has the effect of reducing the toxic and side effects of Western medicine.

[0642] The above shows and describes the basic principles, main features and advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments, and the above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A quality control method for Baibei Yifei Capsules, characterized in that: Including raw material control, preparation method control, identification method and content determination method, as follows: First, raw material control: 1000 single doses of Baibei Yifei Capsules are prepared from the following active ingredients in the following weight ratios: Bletilla striata 118g, Fritillaria thunbergii 40g, Platycodon grandiflorum 67g, Stemona tuberosa 67g, Lilium 67g, Pumice 67g, Aster 67g, Notoginseng 40g, Mahonia laurel 67g, Licorice root 20g; The above nine herbs, including Bletilla striata, Stemona tuberosa, Aster tataricus, Licorice root, Fritillaria thunbergii, Lilium bulb, Panax notoginseng, Platycodon grandiflorum, and Mahonia laurel, should comply with the corresponding medicinal material standards in the Chinese Pharmacopoeia, and Haipumice stone should comply with the corresponding medicinal material standards in the Heilongjiang Provincial Standards for Traditional Chinese Medicines; Second, the preparation method control: Step (1), grinding Bletilla striata, pumice stone, Panax notoginseng and Fritillaria thunbergii into fine powder respectively, sterilizing and setting aside; Step (2), decocting Platycodon grandiflorum, Stemona tuberosa, Lilium bulbiferum, Aster tataricus, Mahonia laurel and Glycyrrhiza uralensis together with water for three times, each time for 1 hour, combining the decoctions, filtering, and concentrating the filtrate under normal pressure into a thick paste having a relative density of greater than or equal to 1.25 at room temperature; the mass of water added in the three decoctions is 6 times, 5 times and 4 times the total mass of the six medicinal materials, respectively, and after the first addition of water, soaking them for 1 hour before decocting; Step (3), mixing the thick paste obtained in step (2) and the fine powder obtained in step (1), drying, crushing, sieving, and filling into capsules to obtain 1000 capsules; Third, identification method: A. Microscopic examination: Take this product and observe it under a microscope: starch granules are oval, broadly oval or elliptical, with a diameter of 6-56μm, and the umbilicus is dot-shaped, crack-shaped, herringbone-shaped or horseshoe-shaped; calcium oxalate needle crystals are bundled or scattered, and the needle crystals are 18-88μm long; B. Thin layer chromatography detection of Panax notoginseng: Take 5g of the content, add 25ml of ethanol, heat and reflux for 30 minutes, filter, evaporate the filtrate, add 20ml of water to dissolve the residue, shake and extract with ether twice, 10ml each time, discard the ether solution, extract twice with water-saturated n-butanol, 10ml each time, combine the n-butanol extracts, wash with 15ml of water-saturated n-butanol, take the n-butanol solution, evaporate to dryness, and dissolve the residue in 2ml of methanol as the test solution; take ginsenoside Rg1 and notoginsenoside R1 reference substances, add methanol to make 1ml of each A solution containing 2 mg of the above solution is used as the reference solution; according to the thin layer chromatography test, 5ul of each of the three solutions is taken and spotted on the same silica gel G thin layer plate, and chloroform, ethyl acetate, methanol and water are mixed in a volume ratio of 15:40:22:10, and then placed below 10°C for stratification, and the lower layer solution is taken as the developing agent, developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots are clearly colored; in the chromatogram of the test product, spots of the same color appear at the corresponding positions of the chromatogram of the reference product; C. Thin layer chromatography detection of Fritillaria thunbergii: Take 5g of the content, add 3ml of concentrated ammonia test solution and 15ml of chloroform, place overnight, filter, take the filtrate and evaporate to dryness, add 1ml of chloroform to the residue to dissolve it, as the test solution; take another reference substance of betel nut A and betel nut B, add chloroform to make a mixed solution containing 2mg of each per 1ml, as the reference substance solution; according to the thin layer chromatography test, take 5-10ul of the test solution and 5ul of the reference substance solution, respectively, and spot them on the same silica gel G thin layer plate, use ethyl acetate-methanol-concentrated ammonia test solution with a volume ratio of 17:2:1 as the developing agent, develop, take out, dry, and spray with dilute potassium bismuth iodide test solution; in the chromatogram of the test sample, at the corresponding position of the chromatogram of the reference substance, a spot of the same color appears; D. Thin layer chromatography detection of licorice: Take 2g of the content, add 20ml of ether, heat and reflux for 1 hour, filter, discard the ether solution, evaporate the residue, add 15ml of methanol, heat and reflux for 1 hour, filter, evaporate the filtrate to dryness, add 20ml of water to dissolve the residue, extract with n-butanol three times, 10ml each time, combine the n-butanol solution, wash with water 3 times, 10ml each time, combine the n-butanol solution, evaporate to dryness, add 1ml of methanol to dissolve the residue, as the test solution; take 1g of licorice control medicinal material, and prepare the control medicinal material solution in the same way; According to the thin layer chromatography test, 2-5ul of the test sample solution and 2ul of the control medicinal material solution were respectively spotted on the same silica gel G thin layer plate prepared with 1% sodium hydroxide solution, and ethyl acetate-formic acid-glacial acetic acid-water with a volume ratio of 15:1:1:2 was used as the developing solvent. The plate was developed, taken out, dried, sprayed with 10% sulfuric acid ethanol solution, and heated at 105°C until the spots were clearly colored. In the chromatogram of the test sample, a main spot of the same color appeared at the corresponding position of the chromatogram of the control medicinal material; E. Thin layer chromatography detection of Bletilla striata: Take 3g of the content, add 20ml of 70% methanol, ultrasonically treat for 30 minutes, filter, evaporate the filtrate, add 10ml of water to the residue to dissolve, shake and extract with ether twice, 20ml each time, combine the ether solution, and evaporate to 1ml as the test solution; take another 1g of Bletilla striata control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, absorb 5-10u1 of the test solution and 5ul of the control medicinal material solution, respectively, and spot them on the same high-efficiency silica gel G thin layer plate, use cyclohexane-ethyl acetate-methanol with a volume ratio of 6:2.5:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, heat at 105℃ for 5-10 minutes, and then leave it for 30-60 minutes; in the chromatogram of the test sample, spots of the same color appear at the corresponding positions of the chromatogram of the control medicinal material; F. Thin layer chromatography detection of Platycodon grandiflorum: Take 7g of the content of this product, add 40ml of methanol, heat and reflux for 30 minutes, cool, filter, evaporate the filtrate to dryness, add 10ml of water to dissolve the residue, add 1ml of hydrochloric acid, heat and reflux for 1 hour, cool, add 30ml of ether to shake and extract, separate the ether solution, wash with 25ml of water, evaporate the ether solution to dryness, and add 1ml of methanol to dissolve the residue as the test solution; take 1g of Platycodon grandiflorum control medicinal material, and prepare the control medicinal material solution in the same way; according to the thin layer chromatography test, take 5-10u1 of the test solution and 5μl of the control medicinal material solution, respectively, and spot them on the same silica gel G thin layer plate, use chloroform-ether with a volume ratio of 1:1 as the developing solvent, develop, take out, dry, spray with 10% sulfuric acid ethanol solution, and heat at 105℃ until the spots are clearly colored; in the chromatogram of the test product, at the corresponding position of the chromatogram of the control medicinal material, a main spot of the same color appears; Fourth, content determination method: Determined by high performance liquid chromatography according to the Chinese Pharmacopoeia; a. Chromatographic conditions and system suitability test: Octadecylsilane bonded silica gel was used as filler; acetonitrile-water with a volume ratio of 18:82 was used as mobile phase; the detection wavelength was 203 nm; the theoretical plate number calculated based on ginsenoside Rg1 should be no less than 3000; b. Preparation of reference solution: Take ginsenoside Rg1 reference substance, add methanol to prepare a solution containing ginsenoside Rg1 0.3 mg per 1 ml, that is; c. Preparation of test solution: Take the contents of 20 pellets, accurately weigh, mix, take 2g, accurately weigh, accurately add 50ml of methanol, weigh, heat and reflux for 1 hour, cool, weigh again, make up the lost weight with methanol, shake well, filter, accurately measure 25ml of the filtrate, evaporate to dryness, add 10ml of water to dissolve, pass through D101 macroporous adsorption resin column, elute with 100ml of water, discard the water; then elute with 100ml of 20% ethanol, discard the 20% ethanol eluate; then elute with 100ml of 95% ethanol, collect the eluate, evaporate to dryness, dissolve the residue with methanol, transfer to a 10ml volumetric flask, add methanol to the scale, shake well, filter, take the filtrate, and obtain; d. Determination method: Accurately pipette 10ul of reference solution and test solution respectively, inject into liquid chromatograph, and determine; Each capsule contains not less than 0.6 mg of Panax notoginseng calculated as ginsenoside Rg1.

2. The quality control method of Baibei Yifei Capsules according to claim 1, characterized in that: In step (1), the particle size of the fine powder is 80-100 mesh; in step (2), filtering is performed using a 80-100 mesh sieve, and the sieve is taken.

3. The quality control method of Baibei Yifei Capsules according to claim 1, characterized in that: In step (3), the sieving is to sieve through a 60-mesh sieve, and the sieve material is taken; and the drying temperature is 70-80°C.

4. The quality control method of Baibei Yifei Capsules according to claim 1, characterized in that: In step c of the content determination method, the inner diameter of the D101 macroporous adsorption resin column is 2 cm and the column length is 15 cm.

5. Use of Baibei Yifei Capsules obtained by the quality control method of Baibei Yifei Capsules according to claim 1 in the preparation of drugs for treating chronic obstructive pulmonary disease.

6. Use of Baibei Yifei Capsules obtained by the quality control method of Baibei Yifei Capsules according to claim 1 in preparing drugs for treating pulmonary fibrosis.

7. Use of Baibei Yifei Capsules obtained by the quality control method of Baibei Yifei Capsules according to claim 1 in preparing drugs for treating pulmonary tuberculosis.

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