CN110426478A

CN110426478A - Tonifying lung capsule quality standard promotes detection method

Info

Publication number: CN110426478A
Application number: CN201910782628.4A
Authority: CN
Inventors: 梁会; 熊静; 王霞; 韩云霞; 杨亮; 蒋玲; 杨仁惠; 屈相玲; 熊成欢; 陈礼大
Original assignee: Second Affiliated Hospital Of Guizhou University Of Traditional Chinese Medicine
Current assignee: Second Affiliated Hospital Of Guizhou University Of Traditional Chinese Medicine
Priority date: 2019-08-23
Filing date: 2019-08-23
Publication date: 2019-11-08

Abstract

The invention discloses a kind of tonifying lung capsule quality standards to promote detection method.The present invention constructs a set of completely new quality standard detecting method, this method is more scientific, the effective component polysaccharide component and Astragaloside IV of main ingredient Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae etc. need to be retained, increase Astragaloside IV and total starches are quantitative Con trolling index, be conducive to that its quality is analysed scientifically and assessed, to improve product quality and ensureing that curative effect provides data support and method validation, and more scientific comprehensive quality standard is established for said preparation.And can get the important content of the effective quality control standard of new Chinese medicine and safety evaluatio, reliable basis is provided for tonifying lung capsule preclinical study, completes the preclinical study of new drug " tonifying lung capsule ".

Description

Tonifying lung capsule quality standard promotes detection method

Technical field

The present invention relates to pharmaceutical field, especially tonifying lung capsule quality standards to promote detection method.

Background technique

Tonifying lung capsule is the second affiliated hospital, Guizhou university of TCM Respiratory Medicine director's Ge Zhenghang chief physician's Empirical formula, It is Preparation in medical units by exploitation, it is clinical use nearly 10 years, curative for effect.We are mainly by Radix Astragali, stir-baked RHIZOMA ATRACTYLODIS MACROCEPHALAE with soil, salt Psoralen The 12 taste Chinese medicinal composition such as rouge, Herba Cistanches, dried orange peel, radix glycyrrhizae preparata.With invigorating the lung and the kidney, relieving cough and asthma effect is mainly used for the lung-distension Disease, the card such as qi deficiency of lung and kidney.We suffer from mainly for the stationary phase (the lung-distension disease qi deficiency of lung and kidney) of Chronic Obstructive Pulmonary Disease (COPD) Person.Original side's proper mass standard falls behind, and primary standard only requires to be studied according to hospital preparation, in initial quality standard, Quan Fang 12 herbal medicines, only 3 qualitative indexes, 1 quantitative target (psoralen, Isopsoralen are assay in psoralea corylifolia), And be non-main ingredient, the requirement of " drug registration management method " (tentative) to 6 kind new medicines is far not achieved, is not enough to reach quality control Purpose.

Chronic Obstructive Pulmonary Disease (COPD) is the chronic respiratory system of a kind of common, multiple, high disability rate and high lethality rate System disease, according to statistics, the people for infecting such disease every year has 600,000,000, and has 2,900,000 people to die of such disease, and lethality occupies the 6th Position, incidence trend is severe, seriously threatens human life and health.Therefore, the study on prevention for carrying out COPD is particularly important.

COPD is the title of a summary, it contains two kinds of main imbalances --- chronic bronchitis and pulmonary emphysema, The common trait of both diseases is all that air can not smoothly circulate lung, and the two is typically co-existing, therefore is united Referred to as chronic obstructive pulmonary disease.Its clinical manifestation is prolonged and repeated cough, expectoration, wheezes and acute respiratory infection occurs, over time Chronic cor pulmonale is evolved into, or even cardiopulmonary failure occurs.Doctor trained in Western medicine generally believes smoking, physical and chemical substance to the thorn of respiratory tract Swash and the factors such as infection are the Etiologicals of COPD^【1】, body influenced by above-mentioned various factors, causes respiratory tract locally anti- Imperial and immune dysfunction, autonomic nervous function imbalance, the tissues such as lung, heart generate pathological change, to show cough, phlegm, asthma Etc. the clinical symptoms of slow Zhi Yan a series of, it is further development of the performance of the cardio-pulmonary functions obstacle such as pulmonary emphysema, pulmonary heart disease.Chinese medicine is recognized For occurring with development often with the invasion repeatedly of exopathogen for COPD, lung, spleen, the dirty functional disturbance of kidney three are closely related.Sick position exists first Lung, exopathogen from mouth and nose, fur invade, mostly first violate lung, lead to impairment of dispersing and descending function of the lung, superinverse and be cough, disturbance in ascending and descending is then asthma.Internal injury in Food, spleen deficiency are unable to transmitting and distributing the fluids, and gathering wet is phlegm, and stain is in lung in damp phlegm.Kidney yang virtual loss, body fluid defeatedization mistake department, lung gasification function lose Often, aqueous vapor is unable to Xuanhua, be phlegm is drink, obstructing airway；Kidney deficiency, vehement, lung saliva of burning in the fire of deficiency type, impairment of dispersing and descending function of the lung, on lung qi It is inverse and cough and asthma is coughed up phlegm.

Anti-inflammatory, antibechic, resolving sputum, the effect relievingd asthma are mainly focused on to the treatment of COPD at present, clinically often combined a variety of The drug of curative effect is treated, but since it is expensive, and toxicity is big, and many patients are difficult to receive.And traditional Chinese medicine has The effective in cure good, advantages such as cheap and easy to get and toxicity is small, for improving COPD patient immune function, slowing down or preventing its lung former Property the development such as heart disease, respiratory failure, pulmonary encephalopathy it is beneficial, it has also become the new issue of research COPD treatment at present.Research report Guilong Kechuanning Capsule has the effect of relieving cough and reducing sputum, depressing Qi and relieving asthma, is clinically used for treating slow Zhi Yan, this belongs to the side of temporary solution；Ginseng Gekko tonifying lung spirit energy help lung spleen kidney, hence it is evident that improve pulmonary ventilation ventilatory, has clear curative effect to chronic obstructive emphysema, this Belong to side of effecting a permanent cure, however still lacks effective kind for the treatment of both manifestation and root cause of disease currently on the market.

Chinese medicine compound prescription tonifying lung capsule is our hospital's cipher prescription, is our province Respiratory Medicine expert --- the positive row religion of chief physician Pueraria lobota The tcm prescription experience awarded forms, and mainly acts on the treatment of Chronic Obstructive Pulmonary Disease (COPD) stationary phase.It is existing in our hospital The applicating history of more than ten years has relatively good clinical efficacy.Tonifying lung capsule mainly for COPD stationary phase patient, with righting Qi-restoratives, kidney and spleen invigorating QI invigorating, lung stomach must be consolidated, to enhance body body fluid and Pulmonary Defense Mechanism, reduce the acute attack period, It prevents the further development of COPD, deteriorate as severe crisis such as cor pulmonale, respiratory failure, pulmonary encephalopathies.Reach and mentions The clinical therapeutic efficacy of high COPD, treating both manifestation and root cause of disease, to improve patients ' life quality, mitigate patient and social economical burden as mesh 's.

China was once used as national emphasis cooperating research class to the prevention and treatment drug research of senile chronic bronchitis Topic, finds the Chinese herbal medicine and compound of not rare significant curative effect, and has carried out the research of basis and application aspect, but be showed no breakthrough Property progress, Major Difficulties prevention and treatment chronic bronchitis acute attack repeatedly and further to pulmonary emphysema, pulmonary artery height Pressure etc., the especially slow scorching every Histopathology index of branch restore normal, i.e., need to be broken through in terms of so-called " effecting a permanent cure ".

Preparation is the crystallization of old docter of TCM's clinical experience in TCM medical organization institute, is the guarantee of tcm clinical practice curative effect, and The embodiment of TCM Features and advantage is the source of new Chinese medicine research and development.Tonifying lung capsule is mainly by Radix Astragali, Radix Codonopsis, Radix Glehniae, benefit The 12 taste Chinese medicinal composition such as bone fat.According to theory of traditional Chinese medical science, symptom shortness of breath lazyness speech, anemophobia be because of lung yin virtual loss caused by, symptom soreness of waist limb Soft, night pollakiuria, dizzy syrigmus is the deficiency of the kidney yin, caused by the failure of the kidney in receiving air because kidney yang is lost.Monarch drug in a prescription Radix Astragali, which has, in we mends The effect of lung QI invigorating, immune enhancing；Ministerial drug Radix Codonopsis, psoralea corylifolia, rhizoma atractylodis macrocephalae, Morinda officinalis, Herba Cistanches, Radix Glehniae etc. are strong with warm kidney The function of essence is mended in spleen tonifying Qi, blood-nourishing；Adjutant dried orange peel, method summer, kadsura longepedunculata, perillaseed have the benefits of preventing phlegm from forming and stopping coughing, nourishiing yin to clear away the lung-heat；Make medicine Radix Glycyrrhizae etc. has the benefits of warming spleen and stomach for dispelling cold, coordinating the drug actions of a prescription.All formulas are matched, treating both manifestation and root cause of disease, plays nourishing lung and kidney, nourishment for vitality, righting altogether The effect of qi-restoratives, lung stomach must be consolidated.The treatment for mainly acting on Chronic Obstructive Pulmonary Disease, lazy speech, fear caused by weak to lung kidney deficiency Wind, soreness of the loins and weakness of the limbs, night pollakiuria, dizziness and tinnitus, body are trembled with fear limb cold, and dry pharynx heat, dysphoria in chestpalms-soles, the diseases such as hectic fever night sweat have well Curative effect.We through thousands of clinical applications in 2002 so far, ten Nian Youyu, and being verified through pharmacodynamics test, tonifying lung capsule energy Improve the FEV0 of COPD rat model_.3、FEV_0.3/ FVC, arterial blood ph, PaO₂, reduce PaCO₂, there is the master for improving lung function Effect is wanted, while having effects that more significant anti-inflammatory, cough-relieving, relievining asthma and resolving sputum, and the immunity of body can be improved, it is seen that it is marked Originally simultaneous to control.COPD stationary phase combines tonifying lung capsule for treating on the basis of western medical treatment, can preferably patients in remission, it is significant to drop Low acute episode frequency improves life quality, has preferable popularization and application value.

Our hospital according to " Guizhou Province's Preparation in medical units drug evaluation main points (Chinese medicine, ethnic drug) " requirement, in 2013 July in year obtains Preparation in medical units authentication code (Guizhou Province medicine word Z20130005), and only the court is using estimated annual output up to 20- 300000 yuan.We by tonifying lung capsule further using the development of six kind new medicines-new Chinese medicine as target, it is close to solve.According in the modern times The specific requirement that the development of medicine compound preparation and new drug are declared, we have been completed tonifying lung capsule part pharmacy and pharmacology side The research in face, technique, quality standard, Pharmacodynamic test of active extract and acute toxicity test including tonifying lung capsule.Due to this quality Standard only requires to be studied according to hospital preparation, in initial quality standard, 12 herbal medicine of Quan Fang, only 3 qualitative indexes, 1 quantitative target (psoralen, Isopsoralen are assay in psoralea corylifolia), and be non-main ingredient, " drug note is far not achieved Volume management method " requirement of (tentative) to 6 kind new medicines, it is not enough to reach quality control purpose.Further to improve declaration material It is required that and effectively control its quality, according to we based on air making-up and spleen enlivening, the effective of main ingredient Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae etc. need to be retained Ingredient polysaccharide component and Astragaloside IV, increase Astragaloside IV and total starches are quantitative Con trolling index.Wish by promoting quality mark Quasi- and long term toxication obtains the important content of the effective quality control standard of new Chinese medicine and safety evaluatio, is tonifying lung capsule Preclinical study provides reliable basis.Complete the preclinical study of new drug " tonifying lung capsule ".

Summary of the invention

The object of the present invention is to provide a kind of tonifying lung capsule quality standards to promote detection method, it can provide a set of completely new Tonifying lung capsule quality standard detection technique, hospital preparation tonifying lung capsule quality standard can be effectively improved by this method.

The present invention is implemented as follows: tonifying lung capsule quality standard promotes detection method, include the following steps:

1) preparation of tonifying lung capsule standard product:

A) by Radix Astragali 229g, Radix Codonopsis 172g, stir-baked RHIZOMA ATRACTYLODIS MACROCEPHALAE with soil 138g, stir-baking RADIX MORINDAE after sprinking salt solution day 138g, stir-baked SEMEN PSORALEAE with salt solution 138g, Herba Cistanches 138g, dried orange peel 103g, rhizoma pinellinae praeparata 103g, roasted perilla fruit 103g, Radix Glehniae 138g, vinegar kadsura longepedunculata 103g and radix glycyrrhizae preparata 57g are mixed It closes uniform；

B) mixing medicinal material is totally submerged with water, and carried out the water for mixing medicinal material and adding 8 times of quality after impregnating 1 hour pan-fried It boils, decocts 2 hours, decoct 3 times altogether, the medical filtration after decoction every time, the filtrate for finally decocting 3 times merges；

C) by the relative density being concentrated under reduced pressure under the conditions of 60 DEG C of filtrate at 60 DEG C be 1.2~1.35 thick paste, then to Silica is added in thick paste to mix；

D) said mixture be dry, pulverize into 60 meshes at 80 DEG C, silica (about 68g) is added and mixes, is packed into glue Capsule is made 1500, obtains tonifying lung capsule standard product；

2) qualitative detection of tonifying lung capsule standard product:

A) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, surpassed Sonication 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, and is shaken with ethyl acetate 30ml It extracts, ethyl acetate layer is evaporated, again with methanol 2ml dissolution, as test solution A；Radix Astragali control medicinal material 1g separately is taken, adds first Alcohol 30ml is ultrasonically treated 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, with acetic acid second Ester 30ml shaking is extracted, and ethyl acetate layer is evaporated, and is dissolved with methanol 1ml, as control medicinal material solution A；According to Chinese Pharmacopoeia The thin-layered chromatography of four general rules 0502 of version in 2015 is tested；Each 5 μ of test solution A and control medicinal material solution A is taken respectively Then l is put respectively on same silica gel g thin-layer plate, using chloroform-acetate-methanol as solvent, chloroform-second Acetoacetic ester-methanol volume ratio is 12:1:1；It spreads out, take out, dry, be subsequently placed in ammonia and smoke 10min, be in wavelength It is inspected under 254nm ultraviolet light on the chromatography position corresponding with the chromatography of control medicinal material solution A of test solution A, if aobvious to have The fluorescence spot of same color；

B) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, surpassed Sonication 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, and is shaken with ethyl acetate 30ml It extracts, ethyl acetate layer is evaporated, and is dissolved with methanol 2ml, as test solution B；Perilla seed control medicinal material powder 1g separately is taken, is added Methanol 30ml is ultrasonically treated 30min, then is filtered, and filtrate is evaporated, and is dissolved with methanol 2ml, as control medicinal material solution B； It is tested according to the thin-layered chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015；3 μ l of test solution B is taken, control is taken 1 μ l of medicinal material solution B puts test solution B and control medicinal material solution B on same silica gel g thin-layer plate, respectively with petroleum ether- Ethyl acetate-glacial acetic acid is solvent, and the boiling point of petroleum ether is 60~90 DEG C, petroleum ether-ethyl acetate-glacial acetic acid volume ratio For 22:2:0.2；It is unfolded, takes out, dries, it is aobvious is heated to spot at 105 DEG C with the ethanol solution of sulfuric acid of 10% mass concentration for spray Color is clear, inspects in the case where wavelength is 365nm ultraviolet light, molten in chromatography and the control medicinal material of test solution B in sample chromatogram On the corresponding position of the chromatography of liquid B, if the aobvious fluorescence spot for having same color；

C) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, surpassed Sonication 30 minutes；It is filtered again, filtrate is evaporated, residue adds water 20ml to make it completely dissolved, and extracts 2 with ether shaking Secondary, each 20ml discards ether liquid, and the n-butanol shaking being saturated with water is extracted 2 times, each 20ml, merges n-butanol liquid, is washed with water It washs 3 times, each 30ml takes n-butanol liquid to be evaporated, and residue adds methanol 2ml to make it completely dissolved, as test solution C；Separately take Control medicinal material solution C is made according to the identical method of test solution C in Radix Glycyrrhizae control medicinal material 1g；According to Chinese Pharmacopoeia 2015 The thin-layered chromatography of four general rules 0502 of version is tested；It takes 5 μ l of test solution C, take 2 μ l of control medicinal material solution C, it will be for Test sample solution C and control medicinal material solution C is taken to be put respectively on same silica gel g thin-layer plate, using chloroform-methanol as solvent, The volume ratio of chloroform and methanol is 9:3；It is unfolded, takes out, dries, sprays with the ethanol solution of sulfuric acid of 10% mass concentration, In 105 DEG C to be heated to spot development clear, inspects in the case where wavelength is 365nm ultraviolet light, in sample chromatogram, in test solution C Chromatography position corresponding with the chromatography of control medicinal material solution C on, if the aobvious fluorescence spot for having same color；

D) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 8g of capsule, add ethyl acetate 50ml, 50 DEG C ultrasonic extraction 1 hour；It is filtered again, concentrates the filtrate to 1ml, as test solution D；Take psoralea corylifolia pair According to medicinal material 0.4g, add ethyl acetate 20ml, is ultrasonically treated 1 hour；It is filtered again, filtrate is evaporated, residue adds ethyl acetate 1ml makes it completely dissolved, as control medicinal material solution D；According to the thin-layer chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015 Method is tested；10~15 μ l of test solution D, control medicinal material solution D 2 μ l are taken, test solution is molten with control medicinal material Liquid D is put respectively on same silica gel g thin-layer plate, and using normal hexane-ethyl acetate as solvent, the volume ratio of normal hexane and acetic acid is 4:1；It is unfolded, takes out, dries, spray is examined with the potassium hydroxide methanol solution of 10% mass concentration in the case where wavelength is 365nm ultraviolet light Depending in sample chromatogram, on the chromatography position corresponding with the chromatography of control medicinal material solution D of test solution D, if aobvious to have The fluorescence spot of same color；

E) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 2g of capsule, add diatomite 2g, grind Carefully, add methanol 25ml, be ultrasonically treated 30 minutes；It is filtered again, after filtrate water bath method, residue adds methanol 1ml to make to dissolve, As test solution E；Dried orange peel control medicinal material 0.5g and diatomite 0.5g separately is taken, according to the preparation method system of test solution E At control medicinal material solution E；It is tested according to the thin-layered chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015；Confession is taken respectively Each 3~5 μ l of test sample solution E and control medicinal material solution E, test solution E and control medicinal material solution E is put respectively in same silica gel On G lamellae, using acetate-methanol-water upper solution as solvent, acetate-methanol-water volume ratio is 6:1: 3；Expansion is taken out, is dried, and spray is inspected in the case where wavelength is 365nm ultraviolet light with alchlor test solution, in sample chromatogram, supplied On the chromatography of test sample solution E position corresponding with the chromatography of control medicinal material solution E, if the aobvious fluorescence spot for having same color；

F) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 2g of capsule, add 70% ethyl alcohol 20ml is impregnated 1 hour, is shaken frequently, is centrifuged, and supernatant, water bath method are taken, and residue adds water 10ml to dissolve, is placed in separatory funnel It is interior, it is extracted with ethyl acetate 2 times, each 10ml is discarded, then is extracted 2 times, each 10ml with n-butanol, and n-butanol liquid, water are merged Bath is evaporated, and residue adds methanol 0.5ml to make to dissolve, as test solution F；Herba Cistanches control medicinal material 1g separately is taken, 30ml water is added to add After heat reflux 30 minutes, centrifugal filtering takes supernatant, sets in separatory funnel, and n-butanol extracts 2 times, and each 20ml merges positive fourth Alcohol liquid, water bath method, residue add methanol 0.5ml to make to dissolve, as control medicinal material solution F；According to Chinese Pharmacopoeia version four in 2015 The thin-layered chromatography of portion's general rule 0502 is tested；Each 2 μ l of test solution F and control medicinal material solution F is drawn, test sample is molten Liquid F and control medicinal material solution F is put respectively on same polyamide film, using methanol-acetic acid-water as solvent, methanol-acetic acid- The volume ratio of water is 2:1:7；It is unfolded, takes out, dries, inspects in the case where wavelength is 365nm ultraviolet light, in sample chromatogram, supplying On the chromatography of test sample solution F position corresponding with the chromatography of control medicinal material solution F, if the aobvious fluorescence spot for having same color.

Related every regulation (Chinese Pharmacopoeia four annex general rules of version in 2015 under [inspection] Ying Fuhe capsule item 0103)。

3) assay

A) Astragaloside IV detects

The preparation of reference substance solution takes Astragaloside IV reference substance appropriate, accurately weighed, adds methanol that every 1ml is made containing 0.3mg Solution to get reference substance solution；

The preparation of test solution: taking at random by intracapsular tolerant 8g, accurately weighed, sets in 100ml centrifuge tube, adds water 30ml, shaking up makes to dissolve, and the n-butanol shaking being saturated with water is extracted 4 times, each 20ml, merges n-butanol liquid, is filled with ammonia solution Divide washing 2 times, each 20mL, then distilled water washing 1 time with 20ml, discards water lotion, merge ammonia washing lotion twice, set on water-bath It waves to no ammonia taste, is saturated with water extracting n-butyl alcohol 2 times, each 20ml, discard washing lotion, merge n-butanol liquid, the recycling of n-butanol liquid To dry, residue adds methanol dissolve and is transferred in 5mL measuring bottle solvent, and methanol is added to shake up, filtration to scale, take subsequent filtrate to get Test solution.

Measuring method is accurate respectively to draw 10 μ l of reference substance solution, 20 μ l, and 20 μ l of test solution injects liquid chromatograph, Measurement is calculated with external standard two-point method logarithmic equation；It is measured according to high performance liquid chromatography general rule 0512；With octadecylsilane key Conjunction silica gel is filler；Using acetonitrile-water as mobile phase, the volume ratio of acetonitrile-water is 32: 68；Evaporative light scattering detector detection, Number of theoretical plate is calculated by Astragaloside IV peak should be not less than 4000；Final every capsule is containing Radix Astragali with Astragaloside IV (C₄₁H₆₈O₁₄) meter, 56 μ g must not be less than；

B) polysaccharide detects

The preparation of reference substance solution takes DEXTROSE ANHYDROUS reference substance appropriate, accurately weighed, adds water that every 1ml is made containing 0.14mg Solution to get reference substance solution；

The preparation of standard curve: precision measures reference substance solution 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml, sets respectively In 15ml tool plug test tube, respectively plus water is mended to 1.0ml, and 4% phenol solution 1ml (facing with configuration) is added in precision, is shaken up, precision plus sulphur Sour 5ml, shakes up, and sets in boiling water bath and heats 20 minutes, takes out, and cooling 5 minutes in ice bath is set, using corresponding reagent as blank, according to purple Outside-visible spectrophotometry general rule 0401 measures absorbance at the wavelength of 485nm, and using absorbance as ordinate, concentration is cross Coordinate draws standard curve；

The preparation of test solution: taking at random by intracapsular tolerant 1g, accurately weighed, is dissolved in water, is transferred to 250ml measuring bottle In, with a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, adds water to scale, shake up, and precision measures 5mL, sets 50ml In centrifuge tube, dehydrated alcohol 20ml is added in precision, shakes up, and refrigeration is stood overnight, and is taken out, centrifugation (revolving speed is 4000 turns per minute) 10 minutes, liquid is discarded supernatant, precipitating plus 80% ethyl alcohol 10ml washed once, and be centrifuged with method, discard supernatant liquid, and precipitating adds water-soluble Solution, is transferred in 50ml measuring bottle, lets cool, add water to scale, shake up to get test solution；

Measuring method: precision measures test solution 1ml, sets in 15ml tool plug test tube, the side under sighting target directrix curve preparation Method is operated from " 4% phenol solution 1ml is added in precision " with method, measures absorbance, and it is molten that test sample is read from standard curve The content of DEXTROSE ANHYDROUS in liquid, calculate to get；Every capsule is containing polysaccharide with DEXTROSE ANHYDROUS (C₆H₁₂O₆) meter, it must not be less than 55mg。

After all of above detection, all detection all passes through, then detects qualification.

In the step c) of step 1), it is 65-70g that silica is added into thick paste.

By adopting the above-described technical solution, the present invention constructs a set of completely new quality standard detecting method, this method It is more scientific, the effective component polysaccharide component and Astragaloside IV of main ingredient Radix Astragali, Radix Codonopsis, Rhizoma Atractylodis Macrocephalae etc. need to be retained, increase Astragaloside IV And total starches are quantitative Con trolling index, are conducive to that its quality is analysed scientifically and assessed, to improve product quality and guarantee Curative effect provides data support and method validation, and establishes more scientific comprehensive quality standard for said preparation.And it can get Chinese medicine The important content of the effective quality control standard of new drug and safety evaluatio, for tonifying lung capsule preclinical study provide reliably according to According to the preclinical study of completion new drug " tonifying lung capsule ".

Detailed description of the invention

Fig. 1 is that Radix Astragali of the invention detects comparative diagram；

Wherein, 1- negative sample, 2- Radix Astragali control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 2 is that perilla seed of the invention detects comparative diagram；

Wherein, 1- negative sample, 2- perilla seed control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 3 is that Radix Glycyrrhizae of the invention detects comparative diagram；

1- negative sample, 2- Radix Glycyrrhizae control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 4 is that psoralea corylifolia of the invention detects comparative diagram；

1- negative sample, 2- psoralea corylifolia control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 5 is that dried orange peel of the invention detects comparative diagram；

1- negative sample, 2- dried orange peel control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 6 is that Herba Cistanches of the invention detect comparative diagram；

1- negative sample, 2- Herba Cistanches control medicinal material, 3~5- tonifying lung capsule test liquid；

Fig. 7 is methanol chromatogram of the invention；

Fig. 8 is Astragaloside IV reference substance chromatogram of the invention；

Fig. 9 is negative control chromatogram of the invention；

Figure 10 is tonifying lung capsule sample chromatogram of the invention；

Figure 11 is Astragaloside IV standard curve of the invention；

Figure 12 is glucose standard curve of the invention.

The invention is simple and feasible, and using effect is good.

Specific embodiment

The embodiment of the present invention: tonifying lung capsule quality standard promotes detection method, includes the following steps:

1) preparation of tonifying lung capsule standard product:

B) mixing medicinal material is totally submerged with water, and after impregnating 1 hour, the water for mixing medicinal material and adding 8 times of quality is carried out pan-fried It boils, decocts 2 hours, decoct 3 times altogether, the medical filtration after decoction every time, the filtrate for finally decocting 3 times merges；

C) by the relative density being concentrated under reduced pressure under the conditions of 60 DEG C of filtrate at 60 DEG C be 1.2~1.35 thick paste, then to Silica is added in thick paste and mixes (about 68g)；

[character] is drafted according to the test result of sample, and test agent is consistent with text description in three batches.This product is ebonite Wafer, content are brown color to chocolate brown powder, and gas is micro-, taste micro-sweet, slightly bitter.

[identification] this product is made of 12 taste Chinese medicines, extracted, refine, and ingredient is more complex.We are in each taste The ingredient of medicinal material has carried out the experimental study of discrimination method, establishes Radix Astragali, perilla seed, Radix Glycyrrhizae, psoralea corylifolia, dried orange peel, Herba Cistanches Thin-layer identification method.

2) detection of tonifying lung capsule standard product:

(1) in tonifying lung capsule Radix Astragali identification

The preparation of test solution: taking this product content 4g, adds methanol 30ml, ultrasonic extraction 30min, and filtering, filtrate is steamed Dry, residue adds water 30ml to make to dissolve, and is shaken and is extracted with ethyl acetate 30ml, and ethyl acetate layer is evaporated, and is dissolved with methanol 2ml, i.e., .

The preparation of negative control solution: Radix Astragali negative control sample 4g is taken, negative control solution is made in the same way of.

The preparation of control medicinal material solution: taking Radix Astragali control medicinal material 1g, adds methanol 30ml, ultrasonic extraction 30min, filters, filter Liquid is evaporated, and residue adds water 30ml to make to dissolve, and is shaken and is extracted with ethyl acetate 30ml, ethyl acetate layer is evaporated, molten with methanol 1ml Solution to get.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws above-mentioned each 5 μ l of three kinds of solution, It is put respectively on same silica gel g thin-layer plate, with chloroform-acetate-methanol (12:1:1) for solvent, is unfolded, takes out, It dries, sets and smoke 10min in ammonia steam, set and inspected under ultraviolet light (254nm).In sample chromatogram, with reference medicine chromatography phase On the position answered, the fluorescence spot of same color is shown.As shown in Figure 1.

(2) in tonifying lung capsule perilla seed identification

The preparation of negative control solution: perilla seed negative control sample 4g is taken, negative control solution is made in the same way of.

The preparation of control medicinal material solution: perilla seed control medicinal material powder 1g is taken, methanol 30ml, ultrasonic extraction 30min, mistake are added Filter, filtrate is evaporated, with methanol 2ml dissolve to get.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws test solution and feminine gender is right According to each 3 μ l of solution, 1 μ l of perilla seed control medicinal material solution, put respectively on same silica gel g thin-layer plate, with petroleum ether (60~90 DEG C)-ethyl acetate-glacial acetic acid (22:2:0.2) be solvent, be unfolded, take out, dry, spray with 10% ethanol solution of sulfuric acid, In 105 DEG C to be heated to spot development clear, sets and inspects under ultraviolet lamp (365nm).In sample chromatogram, with reference medicine chromatography On corresponding position, the fluorescence spot of same color is shown.As shown in Figure 2.

(3) in tonifying lung capsule Radix Glycyrrhizae identification

This product 4g is taken, methanol 30ml is added, ultrasound 30 minutes filters, and filtrate is evaporated, and residue adds water 20ml to make to dissolve, and uses second Ether shaking is extracted 2 times, and each 20ml discards ether liquid.The n-butanol shaking being saturated with water is extracted 2 times, and each 20ml merges positive fourth Alcohol liquid is washed with water 3 times, and each 30ml takes n-butanol liquid to be evaporated, and residue adds methanol 2ml to make to dissolve, as test solution.

The preparation of negative control solution: extracting liquorice negative control sample 4g is made in the same way of negative control solution.

The preparation of control medicinal material solution: extracting liquorice control medicinal material 1g is made in the same way of control medicinal material solution.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws this product test liquid and feminine gender is right According to each 5 μ l of solution, 2 μ l of control medicinal material solution, is put respectively on same silica gel g thin-layer plate, be with chloroform-methanol (9:3) Solvent is unfolded, and takes out, dries, and with 10% ethanol solution of sulfuric acid, it is clear to be heated to spot development at 105 DEG C for spray, sets ultraviolet light Lamp (365nm) is inspected.In sample chromatogram, on position corresponding with reference medicine chromatography, the fluorescence spot of same color is shown. As shown in Figure 3.

(4) in tonifying lung capsule psoralea corylifolia identification

The preparation of test solution: taking this product content 8g, adds ethyl acetate 50ml, 50 DEG C ultrasonic extraction 1 hour, filter It crosses, filtrate is concentrated into 1ml, as test solution.

The preparation of negative control solution: psoralea corylifolia negative control sample 8g is taken, negative control solution is made in the same way of.

The preparation of control medicinal material solution: taking psoralea corylifolia control medicinal material 0.4g, adds ethyl acetate 20ml, is ultrasonically treated 1 hour, Filtration, filtrate are evaporated, and residue adds ethyl acetate 1ml to make to dissolve, as control medicinal material solution.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws test solution and feminine gender is right According to each 10~15 μ l of solution, 2 μ l of control medicinal material solution, put respectively on same silica gel g thin-layer plate, with normal hexane-ethyl acetate (4:1) is solvent, is unfolded, and takes out, dries, and sprays with 10% potassium hydroxide methanol solution, sets and see under ultraviolet lamp (365nm) It examines.In sample chromatogram, on position corresponding with reference medicine chromatography, the spot of same color is shown.As shown in Figure 4.

(5) in tonifying lung capsule dried orange peel identification

The preparation of test solution: taking this product content 2g, adds diatomite 2g, finely ground, adds methanol 25ml, ultrasonic treatment 30 Minute, it filters, after filtrate water bath method, residue adds methanol 1ml to make to dissolve, as test solution.

The preparation of negative control solution: dried orange peel negative control sample 2g is taken, negative control solution is made in the same way of.

The preparation of control medicinal material solution: dried orange peel control medicinal material 0.5g separately is taken, adds diatomite 0.5g, is made in the same way of control medicinal material Solution.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws above-mentioned three kinds of solution each 3~5 μ l puts respectively on same silica gel g thin-layer plate, using the upper solution of acetate-methanol-water (6:1:3) as solvent, opens up It opens, takes out, dry, spray with alchlor test solution, set and observed under ultraviolet lamp (365nm).In sample chromatogram, with comparison medicine Wood color is composed on corresponding position, and identical fluorescence spot is shown.As shown in Figure 5.

(6) in tonifying lung capsule Herba Cistanches identification

The preparation of test solution: taking this product content 2g, adds 70% ethyl alcohol 20ml, impregnates 1 hour, shakes frequently, from The heart, takes supernatant, water bath method, and residue adds water 10ml to dissolve, and is placed in separatory funnel, is extracted with ethyl acetate 2 times, every time 10ml is discarded, then is extracted 2 times, each 10ml with n-butanol, and n-butanol liquid, water bath method are merged, and residue adds methanol 0.5ml to make Dissolution, as test solution.

The preparation of negative control solution: Herba Cistanches negative control sample 2g is taken, negative control solution is made in the same way of.

The preparation of control medicinal material solution: taking Herba Cistanches control medicinal material 1g, after adding 30ml water to be heated to reflux 30 minutes, centrifugation filter It crosses, takes supernatant, set in separatory funnel, n-butanol extracts 2 times, each 20ml, merges n-butanol liquid, water bath method, and residue adds Methanol 0.5ml makes to dissolve, as control medicinal material solution.

It is tested according to thin-layered chromatography (four general rules 0502 of Chinese Pharmacopoeia version in 2015), draws above-mentioned each 2 μ l of three kinds of solution, It is put respectively on same polyamide film, with methanol-acetic acid-water (2:1:7) for solvent, is unfolded, takes out, dry, set ultraviolet It is inspected under light lamp (365nm).In sample chromatogram, on position corresponding with reference medicine chromatography, the spot of same color is shown. As shown in Figure 6.

[inspection] is by the regulation inspection under four general rules of " Chinese Pharmacopoeia " version in 2015,0103 capsule item.Check respectively for water Divide, content uniformity, disintegration time limited, microbial limit.

[moisture] is by four 0832 aquametry of general rule (the second method) measurements of " Chinese Pharmacopoeia " version in 2015.

[content uniformity] is by the regulation inspection under four general rules of " Chinese Pharmacopoeia " version in 2015,0103 capsule item.

[disintegration time limited] is checked by four 0921 disintegration time limit test of general rule regulations of " Chinese Pharmacopoeia " version in 2015.

1 tonifying lung capsule inspection result of table

3) assay

Astragaloside IV is measured according to high performance liquid chromatography (four general rules 0512 of " Chinese Pharmacopoeia " version in 2015).

1 experimental material

(detector is 80 evaporative light of SEDERE LT-ELSD to 3000 type high performance liquid chromatograph of Dionex Μ Ltimate Detector)；Ten a ten thousandth balances (Mei Tele-support benefit AG285 type)；Electric drying oven with forced convection (101-2AB type, Tianjin Stettlen Instrument Ltd.)；It is bright and limpid^TM- D 24UV type pure water system (German Merck millipore company).

Methanol, acetonitrile (chromatographically pure), Shanghai Merck Chemical Engineering Technology Co., Ltd；N-butanol, ammonium hydroxide (analysis is pure), Tianjin Zhi Yuan chemical reagent Co., Ltd；Water is ultrapure water.

Astragaloside IV (110781-200613) is purchased from National Institute for Food and Drugs Control, and content is in terms of 100%.

2 content assaying methods

The preparation of 2.1 reference substance solutions

Precision weighs Astragaloside IV reference substance 4.22mg, sets in 10ml measuring bottle, adds methanol to make to dissolve, and be diluted to scale, It shakes up to get reference substance stock solution.Precision draws reference substance stock solution 2.5ml, adds methanol constant volume to 5ml to get Astragaloside IV Comparison liquid.

The preparation of 2.2 test solutions

This product 8g is taken, it is accurately weighed, it sets in 100ml centrifuge tube, adds water 30ml, shaking up makes to dissolve, the positive fourth being saturated with water Alcohol shaking is extracted 4 times, each 20ml, is merged n-butanol liquid, is sufficiently washed 2 times with ammonia solution, each 20mL, then distilled with 20ml Water washing 1 time, water lotion is discarded, merges ammonia washing lotion twice, sets water-bath Back stroke to no ammonia taste, be saturated with water extracting n-butyl alcohol 2 It is secondary, washing lotion is discarded, merges n-butanol liquid, to doing, residue adds methanol to dissolve and is transferred to 5mL measuring bottle n-butanol liquid recycling design In, add methanol to shake up to scale, filter, take subsequent filtrate to get.

The preparation of 2.3 negative sample solution

This product Radix Astragali negative control sample about 8g is taken, it is accurately weighed, according to the method under the preparation of 2.2 test solutions, From " setting in 100ml centrifuge tube ", with method operate to get.

2.4 chromatographic condition

Agilent Hypersil ODS 5um C18(250×4.6mm)；Mobile phase: acetonitrile-water (32: 68)；Column temperature: 30℃；Flow velocity: 1.0mL/min；Nitrogen pressure: 3.4Pa；Drift tube temperature: 40 DEG C；In theory, the number of plates is no less than 4000。

3 methodological studies

The test of 3.1 specificities

It is accurate respectively to draw Astragaloside IV contrast solution, negative control solution, blank solution and each 20 μ l of test solution Injecting chromatograph is measured by the chromatographic condition under 2.4.See Fig. 7-10.

From map it is found that under this chromatographic condition, Astragaloside IV chromatographic peak is kept completely separate with other components peak, with Radix Astragali At the identical retention time of first glycosides chromatographic peak, negative control is noiseless.

The drafting of 3.2 standard curves

2.1 lower Astragaloside IV reference substance solutions are taken, 5 μ l of sample introduction, 10 μ l, 15 μ l, 20 μ l, 25 μ l, 30 μ l, are pressed respectively 2.4 lower chromatographic conditions are measured.Corresponding peak area is recorded, using peak area logarithm Y as ordinate, with sample volume logarithm Value X is abscissa, draws standard curve and carries out recurrence calculating.The regression equation of Astragaloside IV are as follows: Y=1.5838X+ 0.6927, R²=0.9996.Linear relationship is shown in Table 1-1, and standard curve is shown in Figure 11.

Table 1-1 Specification Curve of Increasing result (n=6)

The result shows that when the mass range of Astragaloside IV is 1.055~6.330 μ g, quality logarithm and peak area logarithm Value has preferable linear relationship.

3.3 precision test

2.1 lower Astragaloside IV reference substance solutions are taken, continuous sample introduction 6 times, the results are shown in Table 1-2.The result shows that RSD is 1.82% (n=6), instrument precision is good.

Table 1-2 Precision test result (n=6)

3.4 repetitive test

This product (lot number 20170302) about 8g is taken, it is accurately weighed, totally 6 parts, test solution is prepared, sample introduction measures, as a result It is shown in Table 1-3.The result shows that the average content of Astragaloside IV is 0.1434mg/g in tonifying lung capsule, RSD is 2.93% (n=6), The result shows that this method repeatability is good.

Table 1-3 repetitive test result (n=6)

3.5 stability test

Take the test solution of serial number 3 under " repetitive test " item, it is closed, be placed at room temperature for, respectively 0h, 6h, 8h, 10h, 12h, it detects under time interval for 24 hours, the results are shown in Table 1-4.Stablize the result shows that test solution is placed at room temperature in for 24 hours.

Table 1-4 stability test result (n=6)

The test of 3.6 sample recovery rates

Taking lot number is 20170302 tonifying lung capsule 's content about 4g, accurately weighed, totally 6 parts, by the 100% of known content Astragaloside IV reference substance solution is added, by 2.2 lower section legal system available test sample solutions, measures in accordance with the law, calculates the rate of recovery.As a result see Table 1-5.The result shows that it is 2.57% (n=6) that the sample recovery rate of Astragaloside IV, which is 103.73%, RSD value, illustrate this method Accuracy is good.

Table 1-5 sample recovery rate test result

The measurement of 4 samples

Tonifying lung capsule 's content about 8g is taken, it is accurately weighed, by 2.2 lower section legal system available test sample solutions, measures, count in accordance with the law Astragaloside content in sample is calculated, the results are shown in Table 1-6.

Determination of Astragaloside result in table 1-6 tonifying lung capsule

It is 70 μ 00 that above-mentioned 3 batches of samples, which measure Astragaloside IV average content,.Consider from mass production, every gram of Radix Astragali first of this product The content of glycosides, limit are calculated with 80%, and the content of Astragaloside IV is 56 μ g/, therefore the limit of this product is set to: every contains Radix Astragali With Astragaloside IV (C₄₁H₆₈O₁₄) meter, 56 μ g/ must not be less than.

Polysaccharide is measured according to ultraviolet spectrophotometry (" Chinese Pharmacopoeia " version general rule 0401 in 2015).

1 experimental material

HP8453 type ultraviolet-uisible spectrophotometer；Ten a ten thousandth balances (Mei Tele-support benefit AG285 type)；SB- 2000 type water-baths (Shanghai Ai Lang Instrument Ltd)；It is bright and limpid^TM- D 24UV type pure water system (German Merck millipore Company), HC-2066 supercentrifuge (An Weizhong Ke Zhongjia scientific instrument Co., Ltd).

Phenol, sulfuric acid, ethyl alcohol (analysis is pure), Chongqing river and mountain Chemical Co., Ltd.；Water is ultrapure water.

D- DEXTROSE ANHYDROUS (0833-9501) is purchased from Chinese pharmaceutical biological product identification institute, and content is in terms of 100%.

2 content assaying methods

The preparation of 2.1 reference substance solutions

Glucose 14.24mg is weighed, distilled water is added to be settled to 100ml to get reference substance solution.

The preparation of 2.2 test solutions

Tonifying lung capsule 's content about 1g is taken, it is accurately weighed, it is dissolved in water, is transferred in 250ml measuring bottle, it is multiple with a small amount of water Container is washed, washing lotion is incorporated in same measuring bottle, adds water to scale, shake up, and precision measures 5mL, sets in 50ml centrifuge tube, and precision adds Enter dehydrated alcohol 20ml, shake up, refrigeration is stood overnight, and is taken out, and centrifugation (revolving speed is 4000 turns per minute) 10 minutes discards supernatant Liquid, precipitating plus 80% ethyl alcohol 10ml washed once, and are centrifuged with method, discard supernatant liquid, precipitating is dissolved in water, and is transferred to 50ml measuring bottle In, let cool, add water to scale, shake up to get.

2.3 measuring method

Precision draws test solution 1ml, sets in 15ml tool plug test tube, and precision is added 4% phenol solution 1ml and (faces with matching Set), it shakes up, then accurate plus sulfuric acid 5ml, shakes up, set in boiling water bath and heat 20 minutes, take out, set cooling 5 minutes in ice bath, with Corresponding reagent is blank, and according to UV-VIS spectrophotometry (general rule 0401), absorbance is measured at the wavelength of 485nm.

3 methodological studies

The preparation of 3.1 standard curves

Precision draws 2.1 lower reference substance solution 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml, sets 15ml tool plug respectively In test tube, respectively plus water is mended to 1.0ml, from " 4% phenol solution 1ml is added in precision ", is developed the color by 2.3 lower section methods, measurement, with Absorbance (A) be ordinate, with concentration of glucose (X) for abscissa, draw standard curve simultaneously carry out linear regression calculating.Grape The regression equation of sugar are as follows: A=0.0632X-0.0310, R²=0.9997, linear relationship table is shown in Table 2-1, and standard curve is shown in Figure 12.

Table 2-1 Specification Curve of Increasing result (n=5)

The result shows that the concentration of glucose is 4.069~12.206 μ g/ml, concentration of glucose and absorbance have preferable Linear relationship.

3.2 precision test

Precision draws reference substance solution 0.4ml, and water is added to mend to 1.0ml, from " 4% phenol solution 1ml is added in precision ", presses 2.3 lower section method colour developings, measurement calculate RSD value, the results are shown in Table 2-2.The result shows that the absorbance values of polysaccharide are 0.454, RSD is 0.11% (n=6), the results showed that, laboratory apparatus precision is good.

Table 2-2 Precision test result (n=6)

3.3 repetitive test

Taking lot number is 20170302 tonifying lung capsule 's content about 1g, accurately weighed, totally 6 parts, prepares test solution, according to Method measurement, calculates RSD value, the results are shown in Table 2-3.The result shows that the average content of polysaccharide is 142.8mg/g, RSD in tonifying lung compound For 1.67% (n=6), the results showed that this method repeatability is good.

Table 2-3 repetitive test result (n=6)

The test of 3.4 sample recovery rates

Taking lot number is 20170302 tonifying lung capsule 's content about 1g, accurately weighed, takes 6 parts altogether, is added by 100% amount D- DEXTROSE ANHYDROUS reference substance solution, measures in accordance with the law, calculates RSD value, the results are shown in Table 2-4.The results show that the sample-adding of polysaccharide is average The rate of recovery is that 99.65%, RSD value is 2.14% (n=6), illustrates that this method accuracy is good.

Table 2-4 sample recovery rate test result (n=6)

3.5 stability test

Taking lot number is 20170302 tonifying lung capsule 's content about 1g, accurately weighed, test sample is prepared in accordance with the law, by 2.3 The colour developing of lower section method, is placed at room temperature for, measures under 0h, 0.5h, 1h, 1.5h, 2h, 2.5h, 3h, 3.5h, 4h time interval respectively, counts RSD value is calculated, the results are shown in Table 2-5.Stablize the result shows that test solution is placed at room temperature in 4h.

Table 2-5 stability test result (n=9)

The measurement of 4 samples

This product content about 1g is taken, it is accurately weighed, assay is carried out to the polysaccharide in 3 batches of tonifying lung capsules in accordance with the law, as a result It is shown in Table 2-6.The result shows that it is 139.61mg/g that above-mentioned 3 batches of samples, which measure polysaccharide average content,.Consider from mass production, this product is every The content of gram polysaccharide, limit are calculated with 80%, and the content of polysaccharide is 69.8mg/, therefore the limit of this product is set to: every contain it is more Sugar (C₆H₁₀O₅) n meter, 55mg must not be less than.

Determination of polysaccharide result in table 2-6 tonifying lung capsule

[specification] every 0.5g.

Claims

1. a kind of tonifying lung capsule quality standard promotes detection method, characterized by the following steps:

1) preparation of tonifying lung capsule standard product:

A) by Radix Astragali 229g, Radix Codonopsis 172g, stir-baked RHIZOMA ATRACTYLODIS MACROCEPHALAE with soil 138g, stir-baking RADIX MORINDAE after sprinking salt solution day 138g, stir-baked SEMEN PSORALEAE with salt solution 138g, Herba Cistanches 138g, Dried orange peel 103g, rhizoma pinellinae praeparata 103g, roasted perilla fruit 103g, Radix Glehniae 138g, vinegar kadsura longepedunculata 103g and radix glycyrrhizae preparata 57g mixing are equal It is even；

B) mixing medicinal material is totally submerged, and after immersion 1 hour with water, adds the water of 8 times of quality to decoct mixing medicinal material, often It is secondary to decoct 2 hours, it decocts 3 times altogether, the medical filtration after decoction, the filtrate for finally decocting 3 times merges；

C) thick paste for being 1.2~1.35 by the relative density being concentrated under reduced pressure under the conditions of 60 DEG C of filtrate at 60 DEG C, then to thick paste Middle addition silica mixes；

D) said mixture be dry, pulverize into 60 meshes at 80 DEG C, silica is added and mixes, and be packed into capsule and be made 1500 Grain obtains tonifying lung capsule standard product；

2) Qualitive test of tonifying lung capsule standard product:

A) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, at ultrasound Manage 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, and is mentioned with ethyl acetate 30ml shaking It takes, ethyl acetate layer is evaporated, again with methanol 2ml dissolution, as test solution A；Radix Astragali control medicinal material 1g separately is taken, adds methanol 30ml is ultrasonically treated 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, and uses ethyl acetate 30ml shaking is extracted, and ethyl acetate layer is evaporated, and is dissolved with methanol 1ml, as control medicinal material solution A；According to Chinese Pharmacopoeia 2015 The thin-layered chromatography of year four general rules 0502 of version is tested；Each 5 μ l of test solution A and control medicinal material solution A is taken respectively, so It is put respectively afterwards on same silica gel g thin-layer plate, using chloroform-acetate-methanol as solvent, chloroform-acetic acid second Ester-methanol volume ratio is 12:1:1；It spreads out, take out, dry, be subsequently placed in ammonia and smoke 10min, be in wavelength It is inspected under 254nm ultraviolet light on the chromatography position corresponding with the chromatography of control medicinal material solution A of test solution A, if aobvious to have The fluorescence spot of same color；

B) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, at ultrasound Manage 30min；It is filtered again, filtrate is evaporated, residue adds water 30ml to make it completely dissolved, and is mentioned with ethyl acetate 30ml shaking It takes, ethyl acetate layer is evaporated, and is dissolved with methanol 2ml, as test solution B；Perilla seed control medicinal material powder 1g separately is taken, adds first Alcohol 30ml is ultrasonically treated 30min, then is filtered, and filtrate is evaporated, and is dissolved with methanol 2ml, as control medicinal material solution B；It presses It is tested according to the thin-layered chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015；3 μ l of test solution B is taken, comparison medicine is taken 1 μ l of material solution B puts test solution B and control medicinal material solution B on same silica gel g thin-layer plate, respectively with petroleum ether-second Acetoacetic ester-glacial acetic acid is solvent, and the boiling point of petroleum ether is 60~90 DEG C, and petroleum ether-ethyl acetate-glacial acetic acid volume ratio is 22:2:0.2；It is unfolded, takes out, dries, sprays with the ethanol solution of sulfuric acid of 10% mass concentration, be heated to spot development at 105 DEG C Clearly, it is inspected in the case where wavelength is 365nm ultraviolet light, in sample chromatogram, in the chromatography and control medicinal material solution of test solution B On the corresponding position of the chromatography of B, if the aobvious fluorescence spot for having same color；

C) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 4g of capsule, add methanol 30ml, at ultrasound Reason 30 minutes；It is filtered again, filtrate is evaporated, residue adds water 20ml to make it completely dissolved, with ether shaking extraction 2 times, often Secondary 20ml discards ether liquid, and the n-butanol shaking being saturated with water is extracted 2 times, each 20ml, merges n-butanol liquid, is washed with water 3 Secondary, each 30ml takes n-butanol liquid to be evaporated, and residue adds methanol 2ml to make it completely dissolved, as test solution C；Another extracting liquorice Control medicinal material solution C is made according to the identical method of test solution C in control medicinal material 1g；According to Chinese Pharmacopoeia version four in 2015 The thin-layered chromatography of portion's general rule 0502 is tested；It takes 5 μ l of test solution C, take 2 μ l of control medicinal material solution C, by test sample Solution C and control medicinal material solution C is taken to be put respectively on same silica gel g thin-layer plate, using chloroform-methanol as solvent, trichlorine The volume ratio of methane and methanol is 9:3；It is unfolded, takes out, dries, sprays with the ethanol solution of sulfuric acid of 10% mass concentration, at 105 DEG C It is clear to be heated to spot development, is inspected in the case where wavelength is 365nm ultraviolet light, in sample chromatogram, in the chromatography of test solution C On position corresponding with the chromatography of control medicinal material solution C, if the aobvious fluorescence spot for having same color；

D) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 8g of capsule, add ethyl acetate 50ml, 50 DEG C ultrasonic extraction 1 hour；It is filtered again, concentrates the filtrate to 1ml, as test solution D；Take psoralea corylifolia control medicinal material 0.4g adds ethyl acetate 20ml, is ultrasonically treated 1 hour；It is filtered again, filtrate is evaporated, residue adds ethyl acetate 1ml to make it It is completely dissolved, as control medicinal material solution D；It is tried according to the thin-layered chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015 It tests；μ l of test solution D10~15,2 μ l of control medicinal material solution D are taken, by test solution and control medicinal material solution D point respectively In on same silica gel g thin-layer plate, using normal hexane-ethyl acetate as solvent, the volume ratio of normal hexane and acetic acid is 4:1；Expansion, It takes out, dry, spray is inspected, test sample with the potassium hydroxide methanol solution of 10% mass concentration in the case where wavelength is 365nm ultraviolet light In chromatography, on the chromatography position corresponding with the chromatography of control medicinal material solution D of test solution D, if aobvious to have same color Fluorescence spot；

E) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 2g of capsule, add diatomite 2g, it is finely ground, Add methanol 25ml, is ultrasonically treated 30 minutes；It is filtered again, after filtrate water bath method, residue adds methanol 1ml to make to dissolve, and makees For test solution E；Dried orange peel control medicinal material 0.5g and diatomite 0.5g separately is taken, is made according to the preparation method of test solution E Control medicinal material solution E；It is tested according to the thin-layered chromatography of Chinese Pharmacopoeia four general rules 0502 of version in 2015；It is taken respectively for examination Product solution E and each 3~5 μ l of control medicinal material solution E, test solution E and control medicinal material solution E is put respectively in same silica G On lamellae, using acetate-methanol-water upper solution as solvent, acetate-methanol-water volume ratio is 6:1: 3；Expansion is taken out, is dried, and spray is inspected in the case where wavelength is 365nm ultraviolet light with alchlor test solution, in sample chromatogram, supplied On the chromatography of test sample solution E position corresponding with the chromatography of control medicinal material solution E, if the aobvious fluorescence spot for having same color；

F) it is sampled from the tonifying lung capsule standard product prepared at random, takes the content 2g of capsule, add 70% ethyl alcohol 20ml, soaked It stain 1 hour, shakes, is centrifuged frequently, take supernatant, water bath method, residue adds water 10ml to dissolve, is placed in separatory funnel, uses second Acetoacetic ester extracts 2 times, and each 10ml is discarded, then is extracted 2 times, each 10ml with n-butanol, merging n-butanol liquid, water bath method, Residue adds methanol 0.5ml to make to dissolve, as test solution F；Herba Cistanches control medicinal material 1g separately is taken, 30ml water is added to be heated to reflux 30 After minute, centrifugal filtering takes supernatant, sets in separatory funnel, and n-butanol extracts 2 times, each 20ml, merges n-butanol liquid, water Bath is evaporated, and residue adds methanol 0.5ml to make to dissolve, as control medicinal material solution F；According to Chinese Pharmacopoeia four general rules of version in 2015 0502 thin-layered chromatography is tested；Draw each 2 μ l of test solution F and control medicinal material solution F, by test solution F and Control medicinal material solution F is put respectively on same polyamide film, using methanol-acetic acid-water as solvent, the body of methanol-acetic acid-water Product is than being 2:1:7；It is unfolded, takes out, dries, is inspected in the case where wavelength is 365nm ultraviolet light, it is molten in test sample in sample chromatogram On the chromatography of liquid F position corresponding with the chromatography of control medicinal material solution F, if the aobvious fluorescence spot for having same color；

3) assay

A) Astragaloside IV detects

The preparation of reference substance solution takes Astragaloside IV reference substance appropriate, accurately weighed, adds methanol that every 1ml is made containing the molten of 0.3mg Liquid is to get reference substance solution；

The preparation of test solution: taking at random by intracapsular tolerant 8g, accurately weighed, sets in 100ml centrifuge tube, adds water 30ml, shake Even to make to dissolve, the n-butanol shaking being saturated with water is extracted 4 times, each 20ml, is merged n-butanol liquid, is sufficiently washed 2 with ammonia solution It is secondary, each 20mL, then distilled water washing 1 time with 20ml, water lotion is discarded, merges ammonia washing lotion twice, sets water-bath Back stroke to nothing Ammonia taste is saturated with water extracting n-butyl alcohol 2 times, each 20ml, discards washing lotion, merges n-butanol liquid, n-butanol liquid recycling design is extremely Dry, residue adds methanol to dissolve and is transferred in 5mL measuring bottle, adds methanol to scale, shakes up, filter, take subsequent filtrate to get test sample Solution；

Measuring method is accurate respectively to draw 10 μ l of reference substance solution, 20 μ l, and 20 μ l of test solution injects liquid chromatograph, measures, It is calculated with external standard two-point method logarithmic equation；It is measured according to high performance liquid chromatography general rule 0512；With octadecylsilane bonded silica Glue is filler；Using acetonitrile-water as mobile phase, the volume ratio of acetonitrile-water is 32: 68；Evaporative light scattering detector detection, it is theoretical Plate number is calculated by Astragaloside IV peak should be not less than 4000；Final every capsule is containing Radix Astragali with Astragaloside IV C₄₁H₆₈O₁₄Meter, must not lack In 56 μ g；

B) polysaccharide detects

The preparation of reference substance solution takes DEXTROSE ANHYDROUS reference substance appropriate, accurately weighed, adds water that every 1ml is made containing the molten of 0.14mg Liquid is to get reference substance solution；

The preparation of standard curve: precision measures reference substance solution 0.2ml, 0.3ml, 0.4ml, 0.5ml, 0.6ml, sets 15ml respectively In tool plug test tube, respectively plus water is mended to 1.0ml, and precision is added 4% phenol solution 1ml, shakes up, and precision plus sulfuric acid 5ml shake up, set It heats 20 minutes, takes out in boiling water bath, cooling 5 minutes in ice bath are set, using corresponding reagent as blank, according to UV-vis spectroscopy light Degree method general rule 0401 measures absorbance at the wavelength of 485nm, and using absorbance as ordinate, concentration is abscissa, draws standard Curve；

The preparation of test solution: taking at random by intracapsular tolerant 1g, accurately weighed, is dissolved in water, is transferred in 250ml measuring bottle, With a small amount of moisture time washing container, washing lotion is incorporated in same measuring bottle, adds water to scale, shake up, and precision measures 5mL, set 50ml from In heart pipe, dehydrated alcohol 20ml is added in precision, shakes up, and refrigeration is stood overnight, and is taken out, centrifugation, and centrifugal rotational speed is per minute 4000 Turn, 10 minutes, discard supernatant liquid, precipitating plus 80% ethyl alcohol 10ml washed once, and be centrifuged with method, discard supernatant liquid, precipitating plus water Dissolution, is transferred in 50ml measuring bottle, lets cool, add water to scale, shake up to get test solution；

Measuring method: precision measures test solution 1ml, sets in 15ml tool plug test tube, the method under sighting target directrix curve preparation, from " precision be added 4% phenol solution 1ml " rises, and operates with method, measures absorbance, from reading nothing in test solution on standard curve The content of water glucose, calculate to get；Every capsule is containing polysaccharide with DEXTROSE ANHYDROUS C₆H₁₂O₆Meter, must not be less than 55mg；

2. tonifying lung capsule quality standard according to claim 1 promotes detection method, it is characterised in that: the step of step 1) C) in, it is 65-70g that silica is added into thick paste.