CN101143164B

CN101143164B - Medicinal composition for treating coronary heart disease and preparation process thereof

Info

Publication number: CN101143164B
Application number: CN2006101521317A
Authority: CN
Inventors: 屠鹏飞; 王亚玲; 李林葆; 王瑛
Original assignee: SHANXI TAIHANG MEDICINE STOCK CO Ltd
Current assignee: SHANXI TAIHANG MEDICINE STOCK CO Ltd
Priority date: 2006-09-14
Filing date: 2006-09-14
Publication date: 2010-12-15
Anticipated expiration: 2026-09-14
Also published as: CN101143164A

Abstract

The invention discloses a drug combination for remedying the coronary heart disease and the preparation method and quality control method of the drug combination. The drug combination consists of an extract of a milkvetch root and the extract of a red salvia root. The preparation method of the drug combination is that the extract of the milkvetch root and the extract of the red salvia root are dissolved in the water for injection; the pH value is regulated; active carbon is added into the solution to be boiled, sealed, frozen, filtered; the filtrate is added with the active carbon to be boiled and filtered; the filtrate is respectively filtered by a microporous filtering film again; the water for injection is added into the filtrate for filling, freezing and drying to obtain the drug combination. The efficacy of drug combination is to benefit qi and activate blood circulation, and the drug combination has obvious effect of remedying the coronary heart disease and especially the angina, caused by the coronary heart disease.

Description

A kind of pharmaceutical composition and preparation method for the treatment of coronary heart disease

Technical field

The present invention relates to a kind of pharmaceutical composition and preparation method thereof and method of quality control, particularly a kind of pharmaceutical composition for the treatment of coronary heart disease and preparation method thereof and method of quality control.

Background technology

Along with the influence of aspect factors such as social population's aging and modern diet, environment, constantly rise based on the sickness rate of the cardiovascular disease of angina pectoris, also receive much attention for the research of cardiovascular disease medicine always.At present, have the medicine of a large amount of treatments and prevention coronary heart disease on the clinical medicine, commonly used mainly contains: the medicine that the expansion blood vessel improves symptom comprises nitrate esters medicine, as nitroglycerin etc., calcium antagonist is as nifedipine, beta-blocker, as propranolol etc., treatment and prophylactic agents such as antiplatelet aggregation are as thrombolytic medicine (as streptokinase, urokinase etc.), anticoagulant (as heparin sodium), antiplatelet drug (as aspirin) etc.But majority is a medicine of alleviating interim symptom, can not play the effect of fundamentally treating angina pectoris, and side effect is also many.Traditional Chinese medical science research thinks that the etiology and pathogenesis of coronary heart disease mainly is a deficiency in origin and excess in superficiality.Deficiency in origin mainly is the deficiency of vital energy, mark is real mainly to be blood stasis, therefore, should be in treatment based on benefiting QI for activating blood circulation, be that the medicine of main method of treatment is more with benefiting QI for activating blood circulation in the present oral formulations, also obtain curative effect preferably, but in for the solid preparation that comparatively is suitable for coronary heart disease treatment, injection type, still do not had the medicine that QI invigorating is invigorated blood circulation now

Summary of the invention

The objective of the invention is to disclose a kind of pharmaceutical composition; Another object of the present invention is to disclose a kind of pharmaceutical composition for the treatment of coronary heart disease; The 3rd purpose of the present invention is this preparation of drug combination method; The 4th purpose of the present invention is the method for quality control of this pharmaceutical composition.

The present invention seeks to be achieved through the following technical solutions:

Pharmaceutical composition of the present invention is made up of following crude drug:

Radix Astragali extract 50～100 weight portions, Radix Salviae Miltiorrhizae extract 80～150 weight portions.

Preferred Radix Astragali extract 60 weight portions, Radix Salviae Miltiorrhizae extract 130 weight portions.

Preferred Radix Astragali extract 90 weight portions, Radix Salviae Miltiorrhizae extract 90 weight portions.

Preferred Radix Astragali extract 75 weight portions, Radix Salviae Miltiorrhizae extract 113 weight portions.

Preferred Radix Astragali extract 50 weight portions, Radix Salviae Miltiorrhizae extract 80 weight portions.

Preferred Radix Astragali extract 100 weight portions, Radix Salviae Miltiorrhizae extract 150 weight portions.

Get the above-mentioned raw materials medicine, add conventional adjuvant, according to common process, be prepared into the dosage form of clinical acceptance, include but are not limited to solid preparations such as tablet, granule, capsule, slow releasing capsule, freeze-dried powder injection, or liquid preparation such as water for injection injection, infusion solutions, oral liquid.

The preparation method of drug combination preparation of the present invention is: get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add the water for injection dissolving of 1500-2500 parts by volume, regulate pH value to 4.5～6.5 with the 5-20% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, to filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again and boiled 10-20 minute, put and be chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, and filtrate adds injects that water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, the water for injection injection is made in fill; Or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

Drug combination preparation preferred for preparation method of the present invention is: get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2000 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5, add 0.5% active carbon with 10% sodium hydroxide solution, 100 ℃ were boiled 30 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, and 80 ℃ were boiled 15 minutes, puts and be chilled to room temperature, filters with the filter paper plate earlier, and filtrate filters with 0.8 μ m and 0.2 μ m microporous filter membrane respectively again, and filtrate adds water and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 2500-3500 parts by volume, water for injection injection or infusion solutions are made in fill, or with filtrate add the injection water complement to 3000 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

Drug combination preparation preferred for preparation method of the present invention is: get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 1500 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5, add 0.3% active carbon with 15% sodium hydroxide solution, 100 ℃ were boiled 20 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.8% active carbon again, 70 ℃ were boiled 40 minutes, put and were chilled to room temperature, filtered with the filter paper plate earlier, filtrate filters with 0.2 μ m and 0.8 μ m microporous filter membrane respectively again, and filtrate adds that the injection water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, the water for injection injection is made in fill, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill; Or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

Drug combination preparation preferred for preparation method of the present invention is: get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2500 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5, add 0.8% active carbon with 5% sodium hydroxide solution, 70 ℃ were boiled 40 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, 90 ℃ were boiled 20 minutes, put and were chilled to room temperature, filtered with the filter paper plate earlier, filtrate filters with 0.6 μ m and 0.4 μ m microporous filter membrane respectively again, and filtrate adds that the injection water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, fill, make the water for injection injection, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

The pass of the above-mentioned weight portion/parts by volume of the present invention is g/ml.Radix Astragali extract, Radix Salviae Miltiorrhizae extract can also can prepare with following extracting method with conventional extraction process preparation of the prior art in the above-mentioned preparation method of the present invention.Radix Astragali total saponins is with astragaloside (C in the Radix Astragali extract of the present invention ₄₁H ₆₈O ₁₄) meter should be not less than 40.0%, contain astragaloside (C ₄₁H ₆₈O ₁₄) should be not less than 40mg/g; Radix Salviae Miltiorrhizae total phenolic acids is with salvianolic acid B (C in the Radix Salviae Miltiorrhizae extract ₃₆H ₃₀O ₁₆) meter should be not less than 60%, salvianolic acid B (C ₃₆H ₃₀O ₁₆) should be not less than 40%.

The preparation method of Radix Astragali extract is: the deionized water that Radix Astragali decoction pieces adds 6-10 times of parts by volume soaked 0.5-1.5 hour, and heating decocts extracted 2-4 hour, filtered filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 1-3 hour, filter; Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 50-70%, cold preservation 6-24 hour to containing the alcohol amount.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10; Concentrated solution injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and it is closely colourless that first deionized water with 10 times of parts by volume is washed till eluent, discards water liquid; 30% ethanol elution of 3-7 times of parts by volume of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection; Reclaim 70% ethanol elution and be concentrated into 50～55 ℃ of relative densities 1.0～1.05, add the water of equimultiple, mixing, cold preservation 6-24 hour, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into 50～55 ℃ of relative densities 1.0～1.05, and lyophilization promptly gets Radix Astragali extract.

The preparation method of Radix Salviae Miltiorrhizae extract is: get salvia piece, add the deionized water of 6-10 times of parts by volume, soaked 0.5-1.5 hour, heating decocts extracted 0.5-1.5 hour, filtered filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 0.5 hour, filter.Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night.Inclining supernatant, the centrifugal filtration of medicinal residues; Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10, centrifugal, supernatant injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and first deionized water with about 15 times of parts by volume is washed till eluent is not had alphanaphthol reaction, discard water elution liquid, reuse 50% ethanol elution is collected 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into 50～55 ℃ of proportions 1.05～1.10, regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization promptly gets Radix Salviae Miltiorrhizae extract.

The method of quality control of injection of the present invention comprises one or more in following discriminating or the assay:

Discrimination method in the method for quality control comprises one or more in the following method:

A, 53% of the compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 69mg-132.5mg, after being dissolved in water, add water saturated n-butanol extraction 1-4 time, merge butanol extraction liquid, with ammonia solution extraction 2-4 time, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10-15: 5-9: the chloroform-methanol-water of 1-5 ratio is placed the lower floor's solution that spends the night below 10 ℃ be developing solvent, launch, take out, dry, spray is with the 5-10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

26.5% of day taking dose of B, the compositions preparation of getting it filled is equivalent to raw material dose 34.5mg-66.3mg,, add methanol and make dissolving,, ultrasonic 5-10 minute, get need testing solution; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 5-10: 1-5: the butyl acetate-formic acid of 1-5 ratio-water is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp of 250-260nm and inspects; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness.

Discrimination method is preferably as follows one or more in the method::

The content 100mg of A, the compositions lyophilized injectable powder of getting it filled is equivalent to raw material dose 100mg, puts in the 50ml triangular flask, after adding the 20ml water dissolution, add water saturated n-butanol extraction 3 times, each 30ml merges butanol extraction liquid, with ammonia solution extraction 3 times, each 20ml discards ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue adds methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

The content 50mg of B, the compositions lyophilized injectable powder of getting it filled is equivalent to raw material dose 50mg, adds methanol 5ml, makes dissolving, and promptly gets need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness.

Content assaying method in the method for quality control of the present invention comprises one or more in the following method::

The assay of A, Radix Astragali total saponins:

The preparation of reference substance solution: it is an amount of that precision takes by weighing the astragaloside reference substance, adds methanol and make the solution that contains 0.25mg among every 1ml;

The preparation of need testing solution: 53% of the compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 69mg-132mg, accurate claim surely, add water and make dissolving, with water saturated n-butanol extraction 2-5 time, merge butanol extraction liquid, with ammonia solution washing 2-4 time, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Algoscopy: precision is measured reference substance solution and each 0.5ml of need testing solution respectively, puts in the color comparison tube, adds the 8% vanillin ethanol solution 0.5ml that faces with newly joining, put in the ice-water bath, add 70-75% sulfuric acid solution 5.0ml, shake up, put in the 60-70 ℃ of water-bath, be incubated 20 minutes, cooling immediately is blank with reagent, measures trap at the 541nm place according to spectrophotography, calculate, promptly;

Drug combination preparation each day taking dose of the present invention is equivalent to raw material dose 188mg, and wherein containing Radix Astragali total saponins is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 12.7-38mg;

The assay of B, astragaloside:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; Evaporative light scattering detector, 40～42 ℃ of drift tube temperatures, carrier gas: nitrogen, nebulizer gas pressure: 2.5Bar, sensitivity 8; Mobile phase: the acetonitrile-water ratio is 20-40: 50-90, flow velocity: 1.0ml/min.Column temperature: 30 ℃, theoretical cam curve is calculated by the astragaloside peak should be not less than 3000;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the astragaloside reference substance, adds methanol and make the solution that every 1ml contains 0.5mg, promptly;

The preparation of need testing solution: get the present invention's 80% of compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 104mg-200mg, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 8-12ml and make dissolving, add 10% ammonia solution 10ml, left standstill water bath method 20-50 minute.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

Algoscopy: accurate respectively reference substance solution 4 μ l, 12 μ l and the need testing solution 20 μ l of drawing, inject chromatograph of liquid, measure peak area, take from right logarithm, the external standard two-point method calculates, promptly;

Drug combination preparation each day taking dose of the present invention is equivalent to raw material dose 188mg, wherein contains Radix Astragali extract with astragaloside (C ₄₁H ₆₈O ₁₄) count 2.52～7.57mg;

The assay of C, salvianolic acid B:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; 10-40: the acetonitrile of 60-90 ratio-0.02% phosphoric acid solution is a mobile phase; The detection wavelength is 288nm.Theoretical cam curve is calculated by the salvianolic acid B peak should be not less than 3000;

The preparation of reference substance solution: it is an amount of that precision takes by weighing the salvianolic acid B reference substance, adds mobile phase and make the solution that every 1ml contains 0.1mg, promptly;

The preparation of need testing solution: 10.6% of the compositions preparation day taking dose of getting it filled is equivalent to raw material dose 13.8mg-26.5mg, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

Algoscopy: accurate respectively reference substance solution and each 20 μ l of need testing solution of drawing, inject chromatograph of liquid, measure, promptly;

Drug combination preparation each day taking dose of the present invention is equivalent to raw material dose 188mg, wherein contains Radix Salviae Miltiorrhizae extract with salvianolic acid B (C ₃₆H ₃₀O ₁₆) meter should be 24.25～68.20mg;

The assay of D, total phenolic acid:

The preparation of reference substance solution: precision is drawn the reference substance solution 2ml under the content of danshinolic acid B mensuration item, puts in the 10ml measuring bottle, and the mobile phase that adds under the content of danshinolic acid B mensuration item is diluted to scale, promptly;

The preparation of need testing solution: the accurate need testing solution 1ml that draws under the salvianolic acid B item, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, as need testing solution;

Algoscopy: get above-mentioned reference substance solution and need testing solution, the mobile phase of measuring under the item with content of danshinolic acid B is blank, measures trap according to spectrophotography at the wavelength place of 288nm, calculates, promptly;

Drug combination preparation each day taking dose of the present invention is equivalent to contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C among the raw material dose 188mg ₃₆H ₃₀O ₁₆) meter should be 64.62～181.75mg.

Assay is preferably as follows one or more in the method:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get test sample lyophilized injectable powder 100mg, the accurate title, decide, and adds water and make dissolving, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Algoscopy: precision is measured reference substance solution and each 0.5ml of need testing solution respectively, puts in the color comparison tube, adds (to face with the 8% vanillin ethanol solution 0.5ml that newly joins, put in the ice-water bath, add 72% sulfuric acid solution 5.0ml, shake up, put in 62 ℃ of water-baths, be incubated 20 minutes, cooling immediately is blank with reagent, measures trap at the 541nm place according to spectrophotography, calculate, promptly;

The every 3ml of pharmaceutical composition freeze-dried powder injection of the present invention, being equivalent to contain Radix Astragali total saponins among the raw material dose 188mg is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 19.0～28.5mg;

The assay of B, astragaloside:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; Evaporative light scattering detector, 40～42 ℃ of drift tube temperatures, carrier gas: nitrogen, nebulizer gas pressure: 2.5Bar, sensitivity 8; Mobile phase: the acetonitrile-water ratio is 33: 67, flow velocity: 1.0ml/min.Column temperature: 30 ℃, theoretical cam curve is calculated by the astragaloside peak should be not less than 3000;

The preparation of need testing solution: get the about 150mg of injection content of the present invention, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 10ml and make dissolving, adds 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

The every 3ml of pharmaceutical composition freeze-dried powder injection of the present invention is equivalent to contain Radix Astragali extract with astragaloside (C among the raw material dose 188mg ₄₁H ₆₈O ₁₄) meter should be 3.79～5.68mg;

The assay of C, salvianolic acid B:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; The acetonitrile of 26: 74 ratios-0.02% phosphoric acid solution is a mobile phase; The detection wavelength is 288nm.Theoretical cam curve is calculated by the salvianolic acid B peak should be not less than 3000;

The preparation of need testing solution: get lyophilized injectable powder test sample 20mg, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

The every 3ml of pharmaceutical composition freeze-dried powder injection of the present invention is equivalent to contain Radix Salviae Miltiorrhizae extract with salvianolic acid B (C among the raw material dose 188mg ₃₆H ₃₀O ₁₆) meter should be 34.25～51.38mg;

The assay of D, total phenolic acid:

The every 3ml of pharmaceutical composition freeze-dried powder injection of the present invention is equivalent to contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C among the raw material dose 188mg ₃₆H ₃₀O ₁₆) meter should be 91.28～136.92mg.

Description of drawings

Fig. 1 astragaloside standard curve

Fig. 2 astragaloside standard curve

Fig. 3 salvianolic acid B standard curve

Fig. 4 salvianolic acid B standard curve

Injection of the present invention is made up of the Radix Astragali, the red sage root two herbal medicines. Main effect is the qi and activate blood circulation effect, the clinical coronary disease and angina pectoris that is used for the treatment of, difference is danshen injections in the past, Clary injection, XUESAITONG ZHUSHEYE, gingko injection (Shu Xuening injection), chuanxiong-rhizome azine injecta, puerarin injections etc. only have the injectable drug of function of promoting blood circulation to disperse blood clots, also be different from astragalus injection, Radix Et Caulis Acanthopanacis Senticosi injections etc. only have the injectable drug of the Chinese medicine of qi invigorating functions, simultaneously again with the effect of present numerous tool qi and activate blood circulation, the oral drugs difference for the treatment of coronary heart disease, injection of the present invention is the drip-feed administration, the emergency and severe disease such to coronary heart disease is also comparatively suitable, through clinical confirmation: the good stability of injection of the present invention, the precision height, therefore the advantages such as reappearance height and average recovery height can bring into play better curative effect.

The content assaying method of astragalus root total saponin is by precision test, records average that the result shows trap and is 0.398, RSD% and be 0.38 result and show that to measure precision better; Test sample study on the stability data result is that 0.420, RSD% is 2.08 for the trap average, and the result shows that need testing solution stablized reappearance test determination trap in 60 minutes, calculate content, the result shows that average is 11.32, RSD%1.73, shows that the method reappearance is better. Application of sample recovery test calculate recovery rate, average % are that 101.51, RSD% is 1.47, and the result shows that the method rate of recovery is higher. By above test explanation, adopt astragalus root total saponin in the red finished product of spectrophotometry stilbene, have method sensitivity, accurately, the advantage such as precision is good, specificity is strong.

About the assay experiment of Astragaloside IV, linear relationship is investigated,, Astragaloside IV concentration is good linear relationship between 0.8128～5.6896 μ g/ μ l as a result. Regression equation: Y=5.4313-1.5011X coefficient correlation, r=0.9996; The Precision test result average is that 8.5294, RSD% is that 1.50 result of the tests show: precision is better; Stability test, average are that 8.5675, RSD% is 0.44, and the result shows need testing solution stability better. The reappearance test, average mg/g is that 28.04, RSD% is 3.22, the result shows that the method reappearance is better. Application of sample recovery test, average recovery rate % are that 101.85, RSD% is 2.57, and the result shows: the rate of recovery is better.

About the assay experiment of tanshin polyphenolic acid B, linear relationship is investigated, and tanshin polyphenolic acid B concentration is good linear relationship in 59.64 ~ 139.16ug/ml scope. Y=19940.66-36617.00X, correlation coefficient r=0.9996. The Precision test result average is that 2384283, RSD% is 0.21, and result of the test shows: precision is better; Stability test, average are that 2386462, RSD% is 0.21, and the result shows need testing solution stability better. The reappearance test, average 29.16, RSD% is 1.99, the result shows that the method reappearance is better. Application of sample recovery test, average recovery rate % are that 101.07, RSD% is 1.66, and the result shows: the rate of recovery is better.

About the assay experiment of total phenolic acid, linear relationship is investigated, and tanshin polyphenolic acid B concentration is linear good relationship in the 9.592-33.57ug/ml scope. Y=0.019921 X-0.00094, correlation coefficient r=0.99997. The Precision test result average is that 0.574, RSD% is 0.21, and result of the test shows: precision is better; Stability test, average are that 0.572, RSD% is 0.84, and the result shows need testing solution stability better. The reappearance test, average 79.37, RSD% is 2.09, the result shows that the method reappearance is better. Application of sample recovery test, average recovery rate % are that 98.05, RSD% is 2.23, and the result shows: the rate of recovery is better.

Following experiment and embodiment are used for further specifying but are not limited to the present invention.

The test of experimental example 1 prescription screening

Table 1 supporting agent is investigated the result

When * calculating content, the support dosage that deduction adds in the total solid

Mainly contain the compositions such as saponins, flavones and glycoside thereof, phenolic acid class because of pharmaceutical composition of the present invention, it is preferably water-soluble that these compositions have, and particularly under the hydrotropy of saponin component, it is water-soluble fine, so do not need to add cosolvent. In addition, saponin component has the effect of supporting agent simultaneously, and in preliminary examination, its skeleton is taken finely, and the dry substance concentration that is easy to freeze-drying and this pharmaceutical composition surpasses 6%, therefore, does not also need to add supporting agent.

Experimental example 2 is about the assay test of astragalus root total saponin ten batch samples

Get the about 100mg of test sample (according to the technical scheme preparation of embodiment 11), accurately weighed, the preparation method makes need testing solution according to need testing solution, measure according to the assay method of drafting, do blank test with method, according to AAS (appendix V of Chinese Pharmacopoeia version in 2000), measure trap at the 541nm place, calculate, and get final product. The results are shown in Table 2.

Table 20 batch sample assay results

According to measurement result, the every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to raw material dose 188mg and contains astragalus root total saponin with Astragaloside IV (C₄₁H ₆₈O ₁₄) meter should be 19.0～28.5mg. By above test explanation, adopt astragalus root total saponin in the red finished product of spectrophotometry stilbene, have method sensitivity, accurately, the advantage such as precision is good, specificity is strong.

Experimental example 3 is about the assay experiment of Astragaloside IV

Take by weighing test sample and make in right amount need testing solution, miillpore filter filters, the accurate 20 μ l that draw, and the injection liquid chromatography is measured, and be get final product. The results are shown in Table 3.

Determination of Astragaloside result in ten batches of pilot products of table 3

According to measurement result, the every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to raw material dose 188mg and contains Astragalus Root P.E with Astragaloside IV (C₄₁H ₆₈O ₁₄) meter should be 3.79～5.68mg. By above test explanation, high performance liquid chromatography-evaporative light detector detects, measure Astragaloside content have the method sensitivity, accurately, quick, the advantage such as precision is good, specificity is strong.

Experimental example 4 is about the assay experiment of tanshin polyphenolic acid B

Instrument and reagent high performance liquid chromatograph: SP8810 pump, SP100 detector, ANSTAR chromatographic work station; Acetonitrile is chromatographically pure (DIKMA company product), and water is the Robust drinking pure water, and phosphoric acid is for analyzing pure (Beijing Chemical Plant).

Chromatographic condition and system suitability octadecyl silane are fixing phase; Acetonitrile-0.02% phosphoric acid (26: 74) is mobile phase.

It is an amount of that precision takes by weighing test sample, gets the freeze drying powder injection of the technical scheme preparation of embodiment 11 and make need testing solution according to the need testing solution preparation method, and miillpore filter filters, the accurate 20 μ l that draw, and the injection liquid chromatography is measured, and calculates, and get final product. Measurement result sees Table 4.

Content of danshinolic acid B measurement result in ten batches of pilot products of table 4

According to measurement result, the every 3ml of the present invention is equivalent to raw material dose 188mg and contains Salvia root P.E with tanshin polyphenolic acid B (C₂₇H ₃₂O ₁₆) meter should be 34.25～51.38mg.

Experimental example 5 is about the assay experiment of total phenolic acid

Instrument and reagent

The tanshin polyphenolic acid B reference substance: it is pure that Chinese pharmaceutical biological product identifies that other agents useful for same of the lot number 111562-2000302 of institute and medicine are analysis.

Instrument model: the WFZ800-D2 of Beijing Analytical Instrument Co., Ltd type ultraviolet-uisible spectrophotometer.

Precision takes by weighing the about 20mg of test sample (according to the technical scheme preparation of embodiment 11), and preparation method's preparation of pressing need testing solution is take corresponding reagent as blank, according to AAS (appendix V of Chinese Pharmacopoeia version in 2000), measure trap at the 288nm place, calculate, and get final product. The results are shown in Table 5.

Table 50 batch sample assay results

According to measurement result, the every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to raw material dose 188mg and contains total phenolic acid in tanshin polyphenolic acid B (C₂₇H ₃₂O ₁₆) should be 91.28～136.92mg.

Following embodiment all can realize the effect of above-mentioned experimental example.

The specific embodiment

Embodiment 1:

Radix Astragali extract 50kg, Radix Salviae Miltiorrhizae extract 150kg make capsule according to a conventional method, every 0.3g, 3 of every days.

Embodiment 2:

Conventional decocting in water alcohol deposition method prepares Radix Astragali extract 100kg, conventional decocting in water alcohol deposition method prepares Radix Salviae Miltiorrhizae extract 80kg, makes tablet according to a conventional method, every 0.3g, 3 of every days.

Embodiment 3:

Conventional decocting in water precipitate with ethanol after the post legal system be equipped with Radix Astragali extract 75kg, conventional decocting in water precipitate with ethanol is equipped with Radix Salviae Miltiorrhizae extract 113kg after post method legal system, makes oral liquid according to a conventional method, every 10ml, 3 of every days.

Embodiment 4:

Conventional ethanol refluxing process prepares Radix Astragali extract 60kg, conventional ethanol refluxing process Radix Salviae Miltiorrhizae extract 113kg, makes injection according to a conventional method.

Embodiment 5:

Get Radix Astragali extract 75kg, Radix Salviae Miltiorrhizae extract 120kg, add adjuvant and make slow releasing capsule according to a conventional method, every 0.2g, every day 2 times.

Embodiment 6:

Get Radix Astragali extract 50kg, Radix Salviae Miltiorrhizae extract 150kg,, add 2000L water for injection and make dissolving, regulate pH value to 4.5～6.5 with 10% sodium hydroxide solution, add 0.5% active carbon, 100 ℃ were boiled 30 minutes, and airtight cold preservation is spent the night; Next day, filter with the filter paper plate, filtrate adds 0.3% active carbon again, and 80 ℃ were boiled 15 minutes, put and were chilled to room temperature, filter with the filter paper plate earlier, filtrate filters with 0.8 μ m and 0.2 μ m microporous filter membrane respectively again, and filtrate adds the injection water and complements to 3000L, fill, lyophilization promptly gets freeze-dried powder injection;

Embodiment 7:

Get Radix Astragali extract 100kg, Radix Salviae Miltiorrhizae extract 80kg, add the water for injection dissolving of 1500L, regulate pH value to 4.5 with 5% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, to filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again and boiled 10-20 minute, put and be chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, filtrate adds the injection water and complements to 30000L and conventional adjuvant, and oral liquid is made in fill.

Embodiment 8:

Get Radix Astragali extract 90kg, Radix Salviae Miltiorrhizae extract 100kg, add the water for injection dissolving of 2000L, regulate pH value to 5 with 15% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, to filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again boiled 10-20 minute, put and was chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, and filtrate adds the injection water and complements to 3000L, fill, freeze-dried powder injection is made in lyophilization.

Embodiment 9:

Get Radix Astragali extract 60kg, Radix Salviae Miltiorrhizae extract 113kg, add the water for injection dissolving of 2200L, regulate pH value to 6 with 10% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again boiled 10-20 minute, put and was chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, and filtrate adds the injection water and complements to 3200L, fill, lyophilization, make freeze-dried powder injection, every 3ml, one of every day.

Embodiment 10:

Get Radix Astragali extract 70kg, Radix Salviae Miltiorrhizae extract 100kg, add the water for injection dissolving of 2200L, regulate pH value to 6 with 10% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again boiled 10-20 minute, put and was chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, filtrate is added the injection water complement to 9500-10500L (parts by volume), fill, make injection, intravenous injection, each 10ml, every day 1 time.

Embodiment 11:

Radix Astragali extract 75kg, Radix Salviae Miltiorrhizae extract 113kg.

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2000ml water for injection and make dissolving, regulate pH value to 4.5～6.5 with 10% sodium hydroxide solution, add 0.5% active carbon, 100 ℃ were boiled 30 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, 80 ℃ were boiled 15 minutes, put and be chilled to room temperature, filter with the filter paper plate earlier, filtrate filters with 0.8 μ m and 0.2 μ m microporous filter membrane respectively again, filtrate adds the injection water and complements to 3000L, fill, lyophilization, freeze-dried powder injection, every 3ml, one of every day.

The wherein preparation of Radix Astragali extract: Radix Astragali decoction pieces adds 8 times of amount deionized waters and soaked 1 hour, and heating decocts extracted 3 hours, filtered filtrate for later use; Medicinal residues add 6 times of amount deionized waters again, decoct and extract 2 times, each 2 hours, filter.Merge 3 times filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 60%, cold preservation 12 hours to containing the alcohol amount.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃).Concentrated solution injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, is washed till eluent closely colourless (water consumption is about 10 times of medical material amount) with deionized water earlier, discards water liquid; 30% ethanol elution of 5 times of amounts of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection.Reclaim 70% ethanol elution and be concentrated into relative density 1.0～1.05 (50～55 ℃), add the water of equimultiple, mixing, cold preservation 12 hours, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into relative density 1.0～1.05 (50～55 ℃), lyophilization promptly gets Radix Astragali extract;

The wherein preparation of Radix Salviae Miltiorrhizae extract: get the salvia piece of recipe quantity, add 8 times of amount deionized waters, soaked 1 hour, heating decocts extracted 1 hour, filtered filtrate for later use; Medicinal residues add 6 times of amount deionized waters again, decoct and extract 2 times, each 0.5 hour, filter.Merge 3 times filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃), centrifugal, supernatant injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, earlier being washed till eluent with deionized water does not have alphanaphthol reaction (about 15 times of water), discards water elution liquid, reuse 50% ethanol elution, collect 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into proportion 1.05～1.10 (50～55 ℃), regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization immediately promptly gets Radix Salviae Miltiorrhizae extract.

Embodiment 12:

Radix Astragali extract 50kg, Radix Salviae Miltiorrhizae extract 80kg

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 1500L water for injection and make dissolving, regulate pH value to 4.5～6.5 with 15% sodium hydroxide solution, add 0.3% active carbon, 100 ℃ were boiled 20 minutes, and airtight cold preservation is spent the night; Next day, filter with the filter paper plate, filtrate adds 0.8% active carbon again, and 70 ℃ were boiled 40 minutes, put and be chilled to room temperature, filter with the filter paper plate earlier, filtrate filters with 0.2 μ m and 0.8 μ m microporous filter membrane respectively again, filtrate is added the injection water complement to 95000-105000L, fill, make infusion solutions, specification 100ml, slow intravenous drip every day.

The wherein preparation of Radix Astragali extract: Radix Astragali decoction pieces adds 6 times of amount deionized waters and soaked 1.5 hours, and heating decocts extracted 2 hours, filtered filtrate for later use; Medicinal residues add 8 times of amount deionized waters again, decoct and extract 1 time, each 3 hours, filter.Merge above filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 60%, cold preservation 12 hours to containing the alcohol amount.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃).Concentrated solution injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, is washed till eluent closely colourless (water consumption is about 10 times of medical material amount) with deionized water earlier, discards water liquid; 30% ethanol elution of 3 times of amounts of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection.Reclaim 70% ethanol elution and be concentrated into relative density 1.0～1.05 (50～55 ℃), add the water of equimultiple, mixing, cold preservation 12 hours, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into relative density 1.0～1.05 (50～55 ℃), lyophilization promptly gets Radix Astragali extract;

The wherein preparation of Radix Salviae Miltiorrhizae extract: get the salvia piece of recipe quantity, add 6 times of amount deionized waters, soaked 1.5 hours, heating decocts extracted 0.5 hour, filtered filtrate for later use; Medicinal residues add 8 times of amount deionized waters again, decoct and extract 1 time, each 0.5 hour, filter.Merge above filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃), centrifugal, supernatant injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, earlier being washed till eluent with deionized water does not have alphanaphthol reaction (about 15 times of water), discards water elution liquid, reuse 50% ethanol elution, collect 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into proportion 1.05～1.10 (50～55 ℃), regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization immediately promptly gets Radix Salviae Miltiorrhizae extract.

Embodiment 13:

Radix Astragali extract 100kg, Radix Salviae Miltiorrhizae extract 150kg add adjuvant and make tablet.

The wherein preparation of Radix Astragali extract: Radix Astragali decoction pieces adds 10 times of amount deionized waters and soaked 0.5 hour, and heating decocts extracted 4 hours, filtered filtrate for later use; Medicinal residues add 4 times of amount deionized waters again, decoct and extract 3 times, each 1 hour, filter.Merge above filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 60%, cold preservation 12 hours to containing the alcohol amount.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃).Concentrated solution injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, is washed till eluent closely colourless (water consumption is about 10 times of medical material amount) with deionized water earlier, discards water liquid; 30% ethanol elution of 7 times of amounts of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection.Reclaim 70% ethanol elution and be concentrated into relative density 1.0～1.05 (50～55 ℃), add the water of equimultiple, mixing, cold preservation 12 hours, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into relative density 1.0～1.05 (50～55 ℃), lyophilization promptly gets Radix Astragali extract;

The wherein preparation of Radix Salviae Miltiorrhizae extract: get the salvia piece of recipe quantity, add 10 times of amount deionized waters, soaked 0.5 hour, heating decocts extracted 1.5 hours, filtered filtrate for later use; Medicinal residues add 4 times of amount deionized waters again, decoct and extract 3 times, each 0.5 hour, filter.Merge above filtrate, be evaporated to relative density 1.05～1.10 (50～55 ℃), be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night.Inclining supernatant, the centrifugal filtration of medicinal residues.Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into relative density 1.05～1.10 (50～55 ℃), centrifugal, supernatant injects in the D101 macroporous resin column of having handled well (ratio of medical material and resin is 1: 3) slowly, earlier being washed till eluent with deionized water does not have alphanaphthol reaction (about 15 times of water), discards water elution liquid, reuse 50% ethanol elution, collect 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into proportion 1.05～1.10 (50～55 ℃), regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization immediately promptly gets Radix Salviae Miltiorrhizae extract.

Embodiment 14:

Radix Astragali extract 75kg, Radix Salviae Miltiorrhizae extract 113kg

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2500L water for injection and make dissolving, regulate pH value to 4.5～6.5 with 5% sodium hydroxide solution, add 0.8% active carbon, 70 ℃ were boiled 40 minutes, and airtight cold preservation is spent the night; Next day, filter with the filter paper plate, filtrate adds 0.3% active carbon again, and 90 ℃ were boiled 20 minutes, put and be chilled to room temperature, filter with the filter paper plate earlier, filtrate filters with 0.6 μ m and 0.45 μ m microporous filter membrane respectively again, filtrate is added the injection water complement to 95000-105000L, add conventional adjuvant, promptly get great transfusion preparation according to common process, every bottle of 100ml of specification, slow intravenous drip every day.

Embodiment 15:

Radix Astragali extract 50kg, Radix Salviae Miltiorrhizae extract 80kg

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2000L water for injection and make dissolving, regulate pH value to 4.5～6.5 with 10% sodium hydroxide solution, add 0.5% active carbon, 100 ℃ were boiled 30 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, 80 ℃ were boiled 15 minutes, put and be chilled to room temperature, filter with the filter paper plate earlier, filtrate filters with 0.8 μ m and 0.2 μ m microporous filter membrane respectively again, filtrate adds the injection water and complements to 3000L, fill, lyophilization promptly gets freeze-dried powder injection, every 3ml, one of every day.

Embodiment 16: discrimination method in the method for quality control

A, get content uniformity and check down content 480mg among the embodiment 1, put in the 50ml triangular flask, add the 20ml water dissolution after, add water saturated n-butanol extraction 3 times, each 30ml, merge butanol extraction liquid, with ammonia solution extraction 3 times, each 20ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

B, get embodiment 2 tablet 240mg, add methanol 5ml, made dissolving, and promptly got need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ 1 of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness.

Embodiment 17: assay in the quality determining method

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get the about 100mg of embodiment 9 lyophilized injectable powder contents, the accurate title, decide, and adds water and make dissolving, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Containing Radix Astragali total saponins among the every 3ml of freeze-dried powder injection of the present invention is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 15.2-22.8mg;

The assay of B, astragaloside:

The preparation of need testing solution: get content 150mg among the embodiment 9, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 10ml and make dissolving, adds 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

Contain Radix Astragali extract with astragaloside (C among every 3ml of freeze-dried powder injection of the present invention ₄₁H ₆₈O ₁₄) meter should be 3.03-4.54mg;

The assay of C, salvianolic acid B:

The preparation of need testing solution: get the about 20mg of content among the embodiment 9, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

Contain Radix Salviae Miltiorrhizae extract with salvianolic acid B (C in every of the freeze-dried powder injection of the present invention ₃₆H ₃₀O ₁₆) meter should be 34.25～51.38mg;

The assay of D, total phenolic acid:

Contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C among every 3ml of freeze-dried powder injection of the present invention ₃₆H ₃₀O ₁₆) meter should be 91.28～136.92mg.

Embodiment 18: quality determining method

Discrimination method is as follows:

A, get content uniformity and check down content 16ml among the embodiment 3, put in the 50ml triangular flask, add the 20ml water dissolution after, add water saturated n-butanol extraction 3 times, each 30ml, merge butanol extraction liquid, with ammonia solution extraction 3 times, each 20ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

B, get the dress embodiment 3 in content 8ml, add methanol 5ml, made dissolving, and promptly got need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Embodiment 19: quality determining method

Discrimination method is as follows:

A, get embodiment 2 tablet 480mg, put in the 50ml triangular flask, add the 20ml water dissolution after, add water saturated n-butanol extraction 3 times, each 30ml, merge butanol extraction liquid, with ammonia solution extraction 3 times, each 20ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

B, get embodiment 2 tablet 240mg, add methanol 5ml, made dissolving, and promptly got need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Embodiment 20: the quality determining method assay is as follows:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get embodiment 15 content 100mg, the accurate title, decide, and adds water and make dissolving, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Containing Radix Astragali total saponins among every 3ml of freeze-dried powder injection of the present invention is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 12.7-19mg;

The assay of B, astragaloside:

The preparation of need testing solution: get embodiment 15 content 150mg, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 10ml and make dissolving, adds 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

Contain Radix Astragali extract with astragaloside (C among every 3ml of freeze-dried powder injection of the present invention ₄₁H ₆₈O ₁₄) meter should be 2.56-3.79mg;

The assay of C, salvianolic acid B:

The preparation of need testing solution: get embodiment 15 content 20mg, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

Contain Radix Salviae Miltiorrhizae extract with salvianolic acid B (C among every 3ml of freeze-dried powder injection of the present invention ₃₆H ₃₀O ₁₆) meter should be 24.25-36.38mg;

The assay of D, total phenolic acid:

Contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C in every 1 of the freeze-dried powder injection of the present invention ₃₆H ₃₀O ₁₆) meter should be 64.62-96.93mg.

Embodiment 21: quality determining method

Assay is as follows:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get embodiment 11 lyophilized injectable powder 100mg, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Containing Radix Astragali total saponins among every 3ml of freeze-dried powder injection of the present invention is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 19.0～28.5mg;

The assay of B, astragaloside:

The preparation of need testing solution: get embodiment 11 lyophilized injectable powder 150mg, put in the 50ml evaporating dish, add methanol 10ml and make dissolving, add 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

Contain Radix Astragali extract among every 3ml of freeze-dried powder injection of the present invention and should be 3.79～5.68mg in astragaloside (C41H68014);

The assay of C, salvianolic acid B:

The preparation of need testing solution: get embodiment 11 lyophilized injectable powder 20mg, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

Contain Radix Salviae Miltiorrhizae extract with salvianolic acid B (C among every 3ml of freeze-dried powder injection ₃₆H ₃₀O ₁₆) meter should be 34.25～51.38mg;

The assay of D, total phenolic acid:

Contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C among every 3ml of freeze-dried powder injection ₃₆H ₃₀O ₁₆) meter should be 91.28～136.92mg.

Embodiment 22: quality determining method

Discrimination method is as follows:

A, get content uniformity and check down content 212mg among the embodiment 5, put in the 50ml triangular flask, add the 20ml water dissolution after, add water saturated n-butanol extraction 3 times, each 30ml, merge butanol extraction liquid, with ammonia solution extraction 3 times, each 20ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

B, get content 106mg among the embodiment 5, add methanol 5ml, made dissolving, and promptly got need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Embodiment 23: quality determining method

Assay is as follows:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get injection 5.3ml among the embodiment 10, the accurate title, decide, and adds water and make dissolving, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

It is C with astragaloside that the every 10ml of injection of the present invention contains Radix Astragali total saponins ₄₁H ₆₈O ₁₄Meter should be 17.73-26.6mg;

The assay of B, astragaloside:

The preparation of need testing solution: get embodiment 10 injection 8ml, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 10ml and make dissolving, adds 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

Contain Radix Astragali extract with astragaloside (C among the every 10ml of injection of the present invention ₄₁H ₆₈O ₁₄) meter should be 3.54-5.30mg;

The assay of C, salvianolic acid B:

The preparation of need testing solution: get embodiment 10 injection 1ml, the accurate title, decide, put in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

The every 10ml of injection of the present invention contains Radix Salviae Miltiorrhizae extract with salvianolic acid B (C ₃₆H ₃₀O ₁₆) meter should be 30.31-45.47mg;

The assay of D, total phenolic acid:

Contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C among the every 10ml of injection of the present invention ₃₆H ₃₀O ₁₆) meter should be 80.78-121.17mg.

Embodiment 24: quality determining method

Discrimination method is as follows:

A, get embodiment 14 infusion solutions 53ml, put in the 100ml triangular flask,, add water saturated n-butanol extraction 3 times, each 30ml merges butanol extraction liquid, with ammonia solution extraction 3 times, and each 20ml, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, placing the lower floor's solution that spends the night below 10 ℃ with the chloroform-methanol-water of 13: 7: 2 ratios is developing solvent, launch, take out, dry, spray is with 10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

B, get embodiment 14 infusion solutions 26.5ml, add methanol 5ml, promptly get need testing solution; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Assay is as follows:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get the 53ml of embodiment 14 infusion solutionses, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml.Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Containing Radix Astragali total saponins among the every 100ml of infusion solutions of the present invention is C with astragaloside ₄₁H ₆₈O ₁₄Meter should be 19.0～28.5mg;

The assay of B, astragaloside:

The preparation of need testing solution: get embodiment 14 infusion solutions 80ml,, put in the 100ml evaporating dish, add methanol 10ml and make dissolving, add 10% ammonia solution 10ml, left standstill water bath method 30 minutes.Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

The every 100ml of infusion solutions of the present invention contains Radix Astragali extract with astragaloside (C ₄₁H ₆₈O ₁₄) meter should be 3.79～5.68mg;

The assay of C, salvianolic acid B:

The preparation of need testing solution: get embodiment 14 infusion solutions 10ml and put water bath method in the 50ml evaporating dish, residue is put in the 25ml measuring bottle, adds the mobile phase dissolving and is diluted to scale, shakes up, the accurate 5ml that draws, put in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

The every 100ml of infusion solutions of the present invention contains Radix Salviae Miltiorrhizae extract with salvianolic acid B (C ₃₆H ₃₀O ₁₆) meter should be 34.25-51.38mg;

The assay of D, total phenolic acid:

The every 100ml of infusion solutions contains Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B (C ₃₆H ₃₀O ₁₆) meter should be 91.28～136.92mg.

Claims

1. detection method for the treatment of the pharmaceutical composition of coronary heart disease, it is characterized in that discrimination method in this method comprise following one or more:

A, 53% of the compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 69mg-132mg, after being dissolved in water, add water saturated n-butanol extraction 1-4 time, merge butanol extraction liquid, with ammonia solution extraction 2-4 time, discard ammoniacal liquor, n-butyl alcohol liquid evaporate to dryness, residue add methanol 2ml makes dissolving, as need testing solution; Other gets the astragaloside reference substance, adds methanol and makes the solution that every 1ml contains 1mg, in contrast product solution; Test according to thin layer chromatography, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 10-15: 5-9: the chloroform-methanol-water of 1-5 ratio is placed the lower floor's solution that spends the night below 10 ℃ be developing solvent, launch, take out, dry, spray is with the 5-10% ethanol solution of sulfuric acid, and it is clear to be heated to the speckle colour developing at 105 ℃; In the test sample chromatograph, with reference substance chromatograph relevant position on, daylight shows down identical sepia speckle;

26.5% of day taking dose of B, the compositions preparation of getting it filled is equivalent to raw material dose 34.5mg-66.3mg, adds methanol and makes dissolving, ultrasonic 5-10 minute, gets need testing solution; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, with 5-10: 1-5: the butyl acetate-formic acid of 1-5 ratio-water is developing solvent, launches, and takes out, and dries, and puts under the ultra-violet lamp of 250-260nm and inspects; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Wherein said pharmaceutical composition crude drug consists of:

Radix Astragali extract 50～100 weight portions, Radix Salviae Miltiorrhizae extract 80～150 weight portions;

Said composition is made tablet, granule, capsule, freeze-dried powder injection solid preparation, or water for injection injection, infusion solutions or oral liquid.

2. detection method for the treatment of the pharmaceutical composition of coronary heart disease, it is characterized in that discrimination method in this method comprise following one or more:

The content 50mg of B, the compositions lyophilized injectable powder of getting it filled is equivalent to raw material dose 50mg, adds methanol 5ml, makes dissolving, and promptly gets need testing solution in ultrasonic 5 minutes; Other gets the salvianolic acid B reference substance, adds methanol and makes the solution that every 1ml contains 2mg, in contrast product solution; According to thin layer chromatography test, draw each 5 μ l of above-mentioned two kinds of solution, put respectively on same silica gel g thin-layer plate, be developing solvent with the butyl acetate-formic acid-water of 7: 2.5: 2.5 ratios, launch, take out, dry, put under the ultra-violet lamp of 254nm and inspect; In the test sample chromatograph, with reference substance chromatograph relevant position on, show identical aubergine speckle; Spray with 3% ferric chloride alcoholic solution after, inspect under the daylight, speckle becomes blueness;

Wherein said pharmaceutical composition crude drug consists of:

3. detection method for the treatment of the pharmaceutical composition of coronary heart disease, it is characterized in that content assaying method in this method comprise following one or more:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: 53% of the compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 69mg-132mg, the accurate title, decide, add water and make dissolving, with water saturated n-butanol extraction 2-5 time, merge butanol extraction liquid, with ammonia solution washing 2-4 time, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml; Filter, discard filtrate just, get subsequent filtrate as need testing solution;

This drug combination preparation each day taking dose is equivalent to contain Radix Astragali total saponins with astragaloside C among the raw material dose 188mg ₄₁H ₆₈O ₁₄Meter should be 12.7-38mg;

The assay of B, astragaloside:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; Evaporative light scattering detector, 40～42 ℃ of drift tube temperatures, carrier gas: nitrogen, nebulizer gas pressure: 2.5Bar, sensitivity 8; Mobile phase: the acetonitrile-water ratio is 20-40: 50-90, flow velocity: 1.0ml/min; Column temperature: 30 ℃, theoretical cam curve is calculated by the astragaloside peak should be not less than 3000;

The preparation of need testing solution: 80% of the compositions preparation day taking dose of getting it filled is equivalent to raw material dose 104mg-200mg, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 8-12ml and make dissolving, add 10% ammonia solution 10ml, left standstill water bath method 20-50 minute; Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

This drug combination preparation each day taking dose is equivalent to contain Radix Astragali extract with astragaloside C among the raw material dose 188mg ₄₁H ₆₈O ₁₄Count 2.52～7.57mg;

The assay of C, salvianolic acid B:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; 10-40: the acetonitrile of 60-90 ratio-0.02% phosphoric acid solution is a mobile phase; The detection wavelength is 288nm; Theoretical cam curve is calculated by the salvianolic acid B peak should be not less than 3000;

The preparation of need testing solution: 10.6% of the compositions preparation day taking dose of getting it filled, be equivalent to raw material dose 13.8mg-26.5mg, the accurate title, decide, and puts in the 25ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, the accurate 5ml that draws puts in the 10ml measuring bottle, add the mobile phase dissolving and be diluted to scale, shake up, 0.45 μ m microporous filter membrane filters, as need testing solution;

This drug combination preparation each day taking dose is equivalent to contain Radix Salviae Miltiorrhizae extract with salvianolic acid B C among the raw material dose 188mg ₃₆H ₃₀O ₁₆Meter should be 24.25～68.20mg;

The assay of D, total phenolic acid:

Algoscopy: get above-mentioned reference substance solution and need testing solution, the mobile phase of measuring under the item with content of danshinolic acid B is blank, measures trap according to spectrophotography at the wavelength place of 288nm, calculates, promptly; This drug combination preparation each day taking dose is equivalent to contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B C among the raw material dose 188mg ₃₆H ₃₀O ₁₆Meter should be 64.62～181.75mg;

Wherein said pharmaceutical composition crude drug consists of:

4. detection method for the treatment of the pharmaceutical composition of coronary heart disease, it is characterized in that content assaying method in this method comprise following one or more:

The assay of A, Radix Astragali total saponins:

The preparation of need testing solution: get test sample lyophilized injectable powder 100mg, the accurate title, decide, and adds water and make dissolving, with water saturated n-butanol extraction 4 times, merge butanol extraction liquid, with ammonia solution washing 3 times, collect n-butyl alcohol liquid evaporate to dryness, residue is with dissolve with methanol and be settled to 50ml; Filter, discard filtrate just, get subsequent filtrate as need testing solution;

Algoscopy: precision is measured reference substance solution and each 0.5ml of need testing solution respectively, puts in the color comparison tube, adds the 8% vanillin ethanol solution 0.5ml that faces with newly joining, put in the ice-water bath, add 72% sulfuric acid solution 5.0ml, shake up, put in 62 ℃ of water-baths, be incubated 20 minutes, cooling immediately is blank with reagent, measures trap at the 541nm place according to spectrophotography, calculate, promptly;

The every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to contain Radix Astragali total saponins with astragaloside C among the raw material dose 188mg ₄₁H ₆₈O ₁₄Meter should be 19.0～28.5mg;

The assay of B, astragaloside:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; Evaporative light scattering detector, 40～42 ℃ of drift tube temperatures, carrier gas: nitrogen, nebulizer gas pressure: 2.5Bar, sensitivity 8; Mobile phase: the acetonitrile-water ratio is 33: 67, flow velocity: 1.0ml/min; Column temperature: 30 ℃, theoretical cam curve is calculated by the astragaloside peak should be not less than 3000;

The preparation of need testing solution: get the about 150mg of this injection content, the accurate title, decide, and puts in the 50ml evaporating dish, adds methanol 10ml and make dissolving, adds 10% ammonia solution 10ml, left standstill water bath method 30 minutes; Residue adds the dissolving of 20% acetonitrile, is transferred in the 25ml measuring bottle, adds 20% dilution in acetonitrile to scale, shakes up, and filters with 0.45 μ m microporous filter membrane, as need testing solution;

The every 3ml of pharmaceutical composition freeze-dried powder injection is equivalent to contain Radix Astragali extract with astragaloside C among the raw material dose 188mg ₄₁H ₆₈O ₁₄Meter should be 3.79～5.68mg;

The assay of C, salvianolic acid B:

Chromatographic condition and system suitability test: with octadecyl silane is immobile phase; The acetonitrile of 26: 74 ratios-0.02% phosphoric acid solution is a mobile phase; The detection wavelength is 288nm; Theoretical cam curve is calculated by the salvianolic acid B peak should be not less than 3000;

The every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to contain Radix Salviae Miltiorrhizae extract with salvianolic acid B C among the raw material dose 188mg ₃₆H ₃₀O ₁₆Meter should be 34.25～51.38mg;

The assay of D, total phenolic acid:

The every 3ml of this pharmaceutical composition freeze-dried powder injection is equivalent to contain Radix Salviae Miltiorrhizae total phenolic acids with salvianolic acid B C among the raw material dose 188mg ₃₆H ₃₀O ₁₆Meter should be 91.28～136.92mg.

Wherein said pharmaceutical composition crude drug consists of:

5. as the detection method of claim 1,2,3 or 4 described pharmaceutical compositions, it is characterized in that the crude drug of the pharmaceutical composition in this method consists of:

Radix Astragali extract 60 weight portions, Radix Salviae Miltiorrhizae extract 130 weight portions;

Or Radix Astragali extract 75 weight portions, Radix Salviae Miltiorrhizae extract 113 weight portions;

Or Radix Astragali extract 90 weight portions, Radix Salviae Miltiorrhizae extract 90 weight portions.

6. as the detection method of claim 1,2,3 or 4 described pharmaceutical compositions, it is characterized in that Radix Astragali extract is prepared by following method in the said composition: the deionized water that Radix Astragali decoction pieces adds 6-10 times of parts by volume soaked 0.5-1.5 hour, heating decocts extracted 2-4 hour, filtered filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 1-3 hour, filter; Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 50-70%, cold preservation 6-24 hour to containing the alcohol amount; Inclining supernatant, the centrifugal filtration of medicinal residues; Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10; Concentrated solution injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and it is closely colourless that first deionized water with 10 times of parts by volume is washed till eluent, discards water liquid; 30% ethanol elution of 3-7 times of parts by volume of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection; Reclaim 70% ethanol elution and be concentrated into 50～55 ℃ of relative densities 1.0～1.05, add the water of equimultiple, mixing, cold preservation 6-24 hour, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into 50～55 ℃ of relative densities 1.0～1.05, and lyophilization promptly gets Radix Astragali extract.

7. as the detection method of claim 1,2,3 or 4 described pharmaceutical compositions, it is characterized in that Radix Salviae Miltiorrhizae extract is prepared by following method in the said composition: get salvia piece, the deionized water that adds 6-10 times of parts by volume, soaked 0.5-1.5 hour, heating decocts extracted 0.5-1.5 hour, filter filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 0.5 hour, filter; Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night; Inclining supernatant, the centrifugal filtration of medicinal residues; Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10, centrifugal, supernatant injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and first deionized water with about 15 times of parts by volume is washed till eluent is not had alphanaphthol reaction, discard water elution liquid, reuse 50% ethanol elution is collected 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into 50～55 ℃ of proportions 1.05～1.10, regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization promptly gets Radix Salviae Miltiorrhizae extract.

8. the detection method of pharmaceutical composition as claimed in claim 5, it is characterized in that Radix Astragali extract is prepared by following method in the said composition: the deionized water that Radix Astragali decoction pieces adds 6-10 times of parts by volume soaked 0.5-1.5 hour, heating decocts extracted 2-4 hour, filtered filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 1-3 hour, filter; Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 50-70%, cold preservation 6-24 hour to containing the alcohol amount; Inclining supernatant, the centrifugal filtration of medicinal residues; Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10; Concentrated solution injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and it is closely colourless that first deionized water with 10 times of parts by volume is washed till eluent, discards water liquid; 30% ethanol elution of 3-7 times of parts by volume of reuse medical material discards water and 30% ethanol elution, and reuse 70% ethanol elution is collected 70% ethanol elution, and does not detect to there being astragaloside with the TLC detection; Reclaim 70% ethanol elution and be concentrated into 50～55 ℃ of relative densities 1.0～1.05, add the water of equimultiple, mixing, cold preservation 6-24 hour, the centrifugal filtration of cold preservation liquid, filtrate is concentrated into 50～55 ℃ of relative densities 1.0～1.05, and lyophilization promptly gets Radix Astragali extract.

9. the detection method of pharmaceutical composition as claimed in claim 5 is characterized in that Radix Salviae Miltiorrhizae extract is prepared by following method in the said composition: get salvia piece, add the deionized water of 6-10 times of parts by volume, soaked 0.5-1.5 hour, heating decocts extracted 0.5-1.5 hour, filtered filtrate for later use; Medicinal residues add the deionized water of 4-8 times of parts by volume again, decoct to extract 1-3 time, each 0.5 hour, filter; Merge above filtrate, be evaporated to 50～55 ℃ of relative densities 1.05～1.10, be cooled to room temperature, add 95% ethanol and reach 70% to containing the alcohol amount, cold preservation is spent the night; Inclining supernatant, the centrifugal filtration of medicinal residues; Merge supernatant and filtrate, decompression recycling ethanol also is concentrated into 50～55 ℃ of relative densities 1.05～1.10, centrifugal, supernatant injects in the D101 macroporous resin column of having handled well slowly, and the ratio of medical material and resin is 1: 3, and first deionized water with about 15 times of parts by volume is washed till eluent is not had alphanaphthol reaction, discard water elution liquid, reuse 50% ethanol elution is collected 50% ethanol elution, until eluent add ferric chloride-iron potassuim cyanide test liquid turn green there is no precipitation till; Reclaim ethanol and be concentrated into 50～55 ℃ of proportions 1.05～1.10, regulate pH value to 4.5～6.5 with 20% sodium hydroxide solution, lyophilization promptly gets Radix Salviae Miltiorrhizae extract.

10. as the detection method of claim 1,2,3 or 4 described pharmaceutical compositions, it is characterized in that this pharmaceutical composition made by following method:

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add the water for injection dissolving of 1500-2500 parts by volume, regulate pH value to 4.5～6.5 with the 5-20% sodium hydroxide solution, add 70-100 ℃ of 0.1-1.0% active carbon and boiled 20-40 minute, airtight cold preservation is spent the night; Next day, to filter, filtrate adds active carbon 70-100 ℃ of 0.1-1.0% again and boiled 10-20 minute, put and be chilled to room temperature, filter, the microporous filter membrane of filtrate reuse 0.1-1.0 μ m filters 1-3 time, and filtrate adds injects that water complements to 30000 parts by volume water and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, fill, make the water for injection injection, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

11. the detection method of pharmaceutical composition as claimed in claim 10 is characterized in that this pharmaceutical composition made by following method:

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2000 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5 with 10% sodium hydroxide solution, add 0.5% active carbon, 100 ℃ were boiled 30 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, 80 ℃ were boiled 15 minutes, put and were chilled to room temperature, filtered with the filter paper plate earlier, filtrate filters with 0.8 μ m and 0.2 μ m microporous filter membrane respectively again, and filtrate adds that the injection water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, fill, make the water for injection injection, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

12. the detection method of pharmaceutical composition as claimed in claim 10 is characterized in that this pharmaceutical composition made by following method:

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 1500 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5 with 15% sodium hydroxide solution, add 0.3% active carbon, 100 ℃ were boiled 20 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.8% active carbon again, 70 ℃ were boiled 40 minutes, put and were chilled to room temperature, filtered with the filter paper plate earlier, filtrate filters with 0.2 μ m and 0.8 μ m microporous filter membrane respectively again, and filtrate adds that the injection water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, fill, make the water for injection injection, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

13. the detection method of pharmaceutical composition as claimed in claim 10 is characterized in that this pharmaceutical composition made by following method:

Get Radix Astragali extract and Radix Salviae Miltiorrhizae extract, add 2500 parts by volume waters for injection and make dissolving, regulate pH value to 4.5～6.5 with 5% sodium hydroxide solution, add 0.8% active carbon, 70 ℃ were boiled 40 minutes, and airtight cold preservation is spent the night; Filter with the filter paper plate next day, and filtrate adds 0.3% active carbon again, 90 ℃ were boiled 20 minutes, put and were chilled to room temperature, filtered with the filter paper plate earlier, filtrate filters with 0.6 μ m and 0.4 μ m microporous filter membrane respectively again, and filtrate adds that the injection water complements to 30000 parts by volume and conventional adjuvant is made oral liquid; Or with filtrate add the injection water complement to the 9500-10500 parts by volume, fill, make the water for injection injection, or with filtrate add the injection water complement to the 95000-105000 parts by volume, infusion solutions is made in fill, or with filtrate add the injection water complement to the 2500-3500 parts by volume, fill, freeze-dried powder injection is made in lyophilization.

14. the detection method of pharmaceutical composition as claimed in claim 5 is characterized in that this pharmaceutical composition made by following method:

15. the detection method of pharmaceutical composition as claimed in claim 14 is characterized in that this pharmaceutical composition made by following method:

16. the detection method of pharmaceutical composition as claimed in claim 14 is characterized in that this pharmaceutical composition made by following method:

17. the detection method of pharmaceutical composition as claimed in claim 14 is characterized in that this pharmaceutical composition made by following method: