CN101084999B

CN101084999B - Compound red sage root freezing-dried powder injection containing notoginseng and its preparation method

Info

Publication number: CN101084999B
Application number: CN 200610014224
Authority: CN
Inventors: 叶正良; 李永强; 郑永锋; 周桂荣
Original assignee: Tianjin Tasly Zhijiao Pharmaceutical Co Ltd
Current assignee: Tianjin Tasly Zhijiao Pharmaceutical Co Ltd
Priority date: 2006-06-08
Filing date: 2006-06-08
Publication date: 2011-09-21
Anticipated expiration: 2026-06-08
Also published as: CN101084999A

Abstract

The present invention provides a compound red sage root freeze dried powder injection containing notoginseng and a preparing process thereof, wherein the injection comprises salvia miltiorrhizae, notoginseng and borneol. The preparing process consists of extracting root of red rooted saliva and pseudo-ginseng extract to obtain red sage root extract, pseudo-ginseng extract, dissolving baras camphor with right amount of ethanol, coating with beta-cyclodextrin, charging injection for water into red sage root extract, notoginseng or red sage root and notoginseng extracts for dissolving, boiling, cooling down, filtering, mixing filtrate with baras camphor beta-CD inclusion solution, charging mannitol, adding right amount of water for injection, finally adjusting pH and freeze-drying. In the freeze dried injection, calculated by the total amount of ginseng saponin Rg1 (C27H32O16), ginseng saponin Rb1 and notoginseng saponin R1, the content of notoginseng active constituent is 12.5-50.0mg/g freeze-dried powder solid.

Description

A kind of compound red sage root freezing-dried powder injection that contains pseudo-ginseng and preparation method thereof

Technical field

The invention belongs to field of medicaments, more particularly, relating to Chinese medicine is that raw material is made a kind of compound red sage root freezing-dried powder injection.

Background technology

In most of western countries, the death of cardiovascular disease accounts for half of total death toll, and wherein coronary heart disease accounts for the first place, accounts for more than 50% of cardiovascular death number.As Asian countries, coronary heart disease is seen not as American-European countries in China more, but in recent years the trend of increasing is arranged.According to the vital statistics data that the Ministry of Public Health announces, the death of nineteen fifty-seven city dweller cardiovascular and cerebrovascular diseases accounts for and rose to 16.6% in total 12.0%, 1989 year, and cause of the death cis-position rises to 2,3 by the 5th, 7.This shows that evidence of coronary heart diseases increases year by year, most of people's cardiovascular risk factors also is ascendant trend continuously and healthily.Patients with coronary heart disease is except causing that miocardial infarction causes the death because of coronary artery blocks, and anginal frequent outbreak also will make its study, ability to work limited greatly, and personal lifestyle can not be taken care of oneself when serious, and family and society have all been brought white elephant.Therefore, since the seventies, one of state key problem has been classified in the preventing and controlling of coronary heart disease as, along with the increase of Incidence of CHD, developing and effectively preventing, to treat coronary heart disease, anginal medicine is a very significant job to reduce angiocardiopathy to the threat of human health.

Modern medicine study is thought, heart is given mechanical irritation do not cause pain, but myocardial ischemia-anoxemia then causes pain.Conflict between the needs of blood supply coronarius and cardiac muscle, coronary blood flow can not satisfy the needs of myocardial metabolism, when causing rapid, temporary transient ischemic of cardiac muscle and anoxic, promptly produces angina pectoris.Modern medicine is the oxygen consumption that improves blood supply coronarius and alleviate cardiac muscle to this sick principle of reatment, treats atherosclerotic simultaneously.Can the use effect during angina pectoris attacks nitric acid preparation faster.Commonly used have monobel 0.3～0.6mg, a sublingual administration; Isosorbide dinitrate 5～10mg, sublingual administration etc.Ill-effect has giddy, distending pain in the head, head beat sense, flushing, palpitaition etc., and idol has blood pressure drops.Prolonged and repeated application can be renderd a service attenuating owing to producing drug resistance.Paracmasis is avoided the various factors that are enough to lead to outbreak that know as far as possible.The antianginal drug that the use effect is lasting, in case angina pectoris attacks, commonly used having (1) nitrate preparations (isosorbide dinitrate, pentaerythritol tetranitrate preparation etc.); (2) adrenergic (inderal, oxprenolol, pindolol, metoprolol etc.), but this class medicine might cause bad reaction, the unexpected inactive possibilities that cause miocardial infarction that have such as postural hypotension; (3) calcium channel blocker (Verapamil, nifedipine etc.), but the danger of excessive inhibition heart is arranged, the improper coronarospasm that might cause of this class medicine of stopping using; (4) coronary artery dilator (persantine, molsidomine, amiodarone etc.), this class medicine especially persantine can reduce side and props up the round-robin volume of blood flow, cause what is called " coronary artery is stolen blood " phenomenon, myocardial ischemia is increased the weight of, though in clinical a lot of arguement commonly used.In addition, the operation of execution ACBG and two kinds of therapeutic method of surgery of percutaneous transluminal coronary angioplasty have also been risen in recent years gradually.But can can operation improve ventricular function, after making severe arrhythmia, heart failure or miocardial infarction do not take place, prolong patient's life-span, still do not have final conclusion; In addition operation itself can concurrent miocardial infarction, but the blood vessel embolism that postoperative is transplanted, so operative indication is strict, is not to be fit to most of patients with coronary heart disease.

Traditional Chinese medicine is the important means of preventing and treating coronary heart disease.At this disease, adopt two methods of taking stopgap measures and effect a permanent cure according to motherland's medical science diagnosis and treatment.Take stopgap measures, mainly use, based on " leading to ", invigorate blood circulation, stagnation resolvation, therapy such as regulate the flow of vital energy, activate yang, reduce phlegm in the pain phase; Effect a permanent cure, generally use, to adjust negative and positive, internal organs, qi and blood, yang-tonifying, enriching yin arranged, fill blood, method such as conditioning viscera in the paracmasis.Wherein commonly used with " activating blood and removing stasis Method " (compound or Chinese patent drug that contains Chinese medicines such as the radix paeoniae rubrathe, the red sage root, safflower, Ligusticum wallichii, cattail pollen, root tuber of aromatic turmeric commonly used) and " aromatic herbs activating yang method " (Suhexian Wan commonly used, Subing drop pills, wide chest ball, heart pill, HMP etc.).But existing Chinese medicine patent medicine is based on decoction and compound pill, and dose is big, carries inconvenience, and onset waits shortcoming slowly.Freeze-dried powder preparation is the advanced formulation of Chinese medicine, and it is slow to overcome the oral formulations onset, can overcome the problem of parenteral solution poor stability again, compare with other formulation, the freeze-dried powder good stability has effectively avoided prescription Chinese traditional medicine generation compatibility to change, and has kept curative effect when prolonging the preparation term of validity; Next technology advanced person, effective substance purity is higher; Also can keep the biologic activity of effective constituent in the compound Danshen Root freeze-dried powder to greatest extent in addition, thereby meet clinical needs, have broad clinical application prospect.

Summary of the invention

The object of the present invention is to provide a kind of compound red sage root freezing-dried powder injection.

Compound red sage root freezing-dried powder injection of the present invention is to be made by following raw materials in part by weight: the red sage root 1000～4000g, pseudo-ginseng 260～850g, borneol 5～50g.

Preferred compound red sage root freezing-dried powder injection of the present invention is to be made by following raw materials in part by weight: the red sage root 2000～3000g, pseudo-ginseng 360～550g, borneol 15～35g.

Best compound red sage root freezing-dried powder injection of the present invention is to be made by following raw materials in part by weight: red sage root 2486.2g, pseudo-ginseng 486.2g, natural borneol 27.6g.

In the medicine of the present invention, the red sage root is a monarch drug in a prescription, promoting blood circulation and removing obstruction in channels, the silt pain relieving of dispelling; Being equipped with pseudo-ginseng is minister, promoting blood circulation and removing blood stasis, remove obstruction in channels to relieve pain; The borneol inducing resuscitation of having one's ideas straightened out, three medicines share, and can reach effect promoting blood circulation and removing blood stasis, regulating qi-flowing for relieving pain.

Effective ingredient of the present invention can adopt following method: water extraction, water extraction and alcohol precipitation method, alcohol extracting-water precipitating, extraction, infusion process, percolation, reflux extraction, continuous backflow extraction method, macroreticular resin absorbing method preparation.

The preparation method of compound red sage root freezing-dried powder injection active principle of the present invention comprises the steps:

A. it is standby to get the above-mentioned recipe quantity red sage root, pseudo-ginseng, borneol;

B. above three flavors, red sage root boiling, collecting decoction, and concentrate, concentrate adds ethanol, leaves standstill, and filters, and filtrate concentrates, concentrate adds ethanol, leaves standstill, and filters, and filtrate concentrates, and adds water, and refrigeration filters, filtrate concentrates, and adds ethanol, leaves standstill, and filters, and filtrate concentrates, and drying gets Salvia root P.E;

Pseudo-ginseng, boiling, collecting decoction concentrates, and adds ethanol, leaves standstill, filter, filtrate concentrates, and adds water, and refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-, add water, heating makes dissolving, adds activated charcoal and boils, and takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, inclusion filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution, promptly.

In the compound lyophilized powder of the present invention, contain Salvia root P.E, Notogineng Extract and borneol.

In the compound red sage root freezing-dried powder injection of the present invention, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 6.25～56.25mg/g freeze-dried powder solid content.

In the preferred compound red sage root freezing-dried powder injection of the present invention, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), ginsenoside Rb1, notoginsenoside R three's total content is 12.5～50.0mg/g freeze-dried powder solid content.

In the best compound red sage root freezing-dried powder injection of the present invention, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), ginsenoside Rb1, notoginsenoside R three's total content is 18.75～31.25mg/g freeze-dried powder solid content.

In the compound red sage root freezing-dried powder injection of the present invention, the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 2.5～25.5mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁3.2～24.5mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁0.55～6.25mg/g freeze-dried powder solid content.

In the preferred compound red sage root freezing-dried powder injection of the present invention, the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 4.5～19.5mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁4.2～18.5mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁0.85～4.05mg/g freeze-dried powder solid content.

In the best compound red sage root freezing-dried powder injection of the present invention, the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 8.076 ± 1.21mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁8.082 ± 1.32mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁2.019 ± 1.17mg/g freeze-dried powder solid content.

Compound red sage root freezing-dried powder injection of the present invention, wherein the red sage root is with danshensu (C ₉H ₁₀O ₅) meter, content is 1.25～18.75mg/g freeze-dried powder solid content.

Compound red sage root freezing-dried powder injection of the present invention, wherein the red sage root is with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 12.5～75.0mg/g freeze-dried powder solid content.

Compound red sage root freezing-dried powder injection of the present invention, wherein borneol is with d-Bomeol (C ₁₀H ₁₈O) meter, content is 6.25～75.0mg/g freeze-dried powder solid content.

Compound red sage root freezing-dried powder injection of the present invention, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 6.25～56.25mg/g freeze-dried powder solid content; The red sage root is with danshensu (C ₉H ₁₀O ₅) meter, content is 1.25～18.75mg//g freeze-dried powder solid content; The red sage root is with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 6.25～100.0mg/g freeze-dried powder solid content; Borneol is with d-Bomeol (C ₁₀H ₁₈O) meter, content is 6.25～75.0mg/g freeze-dried powder solid content.

Preferred compound red sage root freezing-dried powder injection, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 12.5～50.0mg/g freeze-dried powder solid content; The red sage root is with danshensu (C ₉H ₁₀O ₅) meter, content is 2.5～15.0mg/g freeze-dried powder solid content; The red sage root is with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 12.5～75.0mg/g freeze-dried powder solid content; Borneol is with d-Bomeol (C ₁₀H ₁₈O) meter, content is 12.5～562.5mg/g freeze-dried powder solid content.

Preferred compound red sage root freezing-dried powder injection is characterized in that, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 18.75～31.25mg/g freeze-dried powder solid content; The red sage root is with danshensu (C ₉H ₁₀O ₅) meter, content is 3.75～12.75mg/g freeze-dried powder solid content; The red sage root is with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 25.0～50.125mg/g freeze-dried powder solid content; Borneol is with d-Bomeol (C ₁₀H ₁₈O) meter, content is 25.0～50.125mg/g freeze-dried powder solid content.

Best compound red sage root freezing-dried powder injection, wherein pseudo-ginseng is with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 18.75～25.0mg/g freeze-dried powder solid content; The red sage root is with danshensu (C ₉H ₁₀O ₅) meter, content is 3.75～8.875mg/g freeze-dried powder solid content; The red sage root is with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 25.0～41.375mg/g freeze-dried powder solid content; Borneol is with d-Bomeol (C ₁₀H ₁₈O) meter, content is 31.25～41.56mg/g freeze-dried powder solid content.

In the compound lyophilized powder of the present invention, pseudo-ginseng is with the ginsenoside Rg1, the ginsenoside Rb1, and notoginsenoside R total amount meter, content is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D).

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica, are moving phase with the acetonitrile-water gradient elution; The detection wavelength is 203nm.Number of theoretical plate calculates by the ginsenoside Rg1 peak should be not less than 2000.

The linear gradient elution proportion of mobile phase changes program

The preparation of reference substance solution respectively precision to take by weighing ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R reference substance an amount of, add methyl alcohol respectively and make the solution that every 1ml contains 0.5mg, 0.5mg, 0.2mg, shake up, promptly.

This product content under the content uniformity item is got in the preparation of need testing solution, and mixing is got 0.5g, accurate claims surely, puts in the 10ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, and miillpore filter (0.45um) filters, promptly.

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and calculate, promptly.

This product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 6.25～56.25mg/g freeze-dried powder solid content.

In the compound lyophilized powder of the present invention, the red sage root is measured according to high performance liquid chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 D) in danshensu content.

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica; Acetonitrile-0.1% phosphoric acid (8: 92) is moving phase; The detection wavelength is 280nm.Number of theoretical plate calculates by the Sodium Danshensu peak should be not less than 3000.

It is an amount of that the preparation precision of reference substance solution takes by weighing the Sodium Danshensu reference substance, adds methyl alcohol and make the solution that every 1ml contains 0.1mg, shakes up, promptly.

This product content under the content uniformity item is got in the preparation of need testing solution, and mixing is got 0.4g, accurate claims surely, puts in the 25ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, and miillpore filter (0.45um) filters, promptly.

Accurate reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and promptly get (every 1mg Sodium Danshensu is equivalent to the 0.880mg danshensu).

This product contains the red sage root with danshensu (C ₉H ₁₀O ₅) meter, content is 1.25～18.75mg/g freeze-dried powder solid content.

In the compound lyophilized powder of the present invention, the red sage root is in tanshin polyphenolic acid B, and content is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D).

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica, are moving phase with acetonitrile-0.1% phosphoric acid (24: 76); The detection wavelength is 288nm.Number of theoretical plate calculates by the tanshin polyphenolic acid B peak should be not less than 3000.

It is an amount of that the preparation precision of reference substance solution takes by weighing the tanshin polyphenolic acid B reference substance, adds methyl alcohol and make the solution that every 1ml contains 0.15mg, shakes up, promptly.

This product content under the content uniformity item is got in the preparation of need testing solution, and mixing is got 0.15g, accurate claims surely, puts in the 50ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, and miillpore filter (0.45um) filters, promptly.

This product contains the red sage root with tanshin polyphenolic acid B (C ₃₆H ₃₀O ₁₆) meter, content is 12.5～75.0mg/g freeze-dried powder solid content.

In the compound freeze-dried powder of the present invention, content of bornyl alcohol is in d-Bomeol, and content is measured according to vapor-phase chromatography (an appendix VI of Chinese Pharmacopoeia version in 2005 E).

Chromatographic condition and system suitability test chromatographic column: INNOWAX (30m*0.32mm*0.5um), column temperature: temperature programme: 150 ℃ keep 7min, rise to 220 ℃ with 20 ℃/min, keep 5min.Vapourizing temperature: 250 ℃; Detector temperature: 250 ℃; Detecting device: FID flame ionization ditector, hydrogen flowing quantity: 40ml/min; Air mass flow: 350ml/min; Carrier gas: high-purity nitrogen; Carrier gas flux: 2ml/min, tail blows 30ml/min; Input mode: split ratio 5: 1.Number of theoretical plate calculates by the d-Bomeol peak should be not less than 1900.

It is an amount of that the mensuration of correction factor is got gaultherolin, and accurate the title decides, and adds the solution that absolute ethyl alcohol is mixed with 1mg/ml, as inner mark solution.It is an amount of to get the d-Bomeol reference substance, and accurate the title decides, and adds the solution that absolute ethyl alcohol is made 1mg/ml, precision is measured 0.5ml in the 10ml measuring bottle again, is diluted to scale with absolute ethyl alcohol behind the accurate adding inner mark solution 1ml, shakes up, draw 1ul, inject gas chromatograph, the calculation correction factor.

Determination method is got 5 bottles of this product, and inclining content, mixing, precision takes by weighing in right amount (being equivalent to 1 bottle amount approximately), in the 25ml measuring bottle, after the water 5ml dissolving, be diluted to scale with absolute ethyl alcohol, shake up, precision is measured 0.5ml in the 10ml measuring bottle again, be diluted to scale with absolute ethyl alcohol behind the accurate adding inner mark solution 1ml, shake up, filter, draw subsequent filtrate 1ul, inject gas chromatograph is measured, promptly.

This product contains borneol with d-Bomeol (C for every bottle ₁₀H ₁₈O) meter, content is 6.25～75.0mg/g freeze-dried powder solid content.

Medicine of the present invention can adopt the preparation of Chinese medicine preparation conventional method.Also these bulk drugs can be ground into powder evenly makes powder and takes after mixing it with water; Also can be with these medicines decocting together, the condensed water decocting liquid is made oral liquid then; Also can the raw materials used medicinal powder of medicine of the present invention is broken, bulk drug ground ingredients and auxiliary material are mixed direct tablet compressing.The medicine activity component that the present invention can adopt the said extracted method to obtain is made said formulation on any pharmacy with present common medicament theory with auxiliary material; As tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck dosage forms such as agent, granule, electuary, pill, powder, paste, sublimed preparation, supensoid agent, solution, ointment, emplastrum, spray, dripping pill, soft capsule, parenteral solution, freeze drying powder injection, but preferably adopt following method to extract bulk drug, and be prepared into freeze drying powder injection, below just further specify, but this is not construed as limiting the invention of the present invention.

Compound red sage root freezing-dried powder injection of the present invention, its preparation method is as follows:

A. it is standby to get the recipe quantity red sage root, pseudo-ginseng, borneol;

B. the above red sage root, pseudo-ginseng be through extracting or close extraction respectively, Salvia root P.E, Notogineng Extract or the red sage root, pseudo-ginseng are closed extract; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Salvia root P.E, Notogineng Extract or Salvia miltiorrhiza and Panax notoginseng extract are added the injection water make dissolving in right amount, boil, put coldly, filter, filtrate is mixed with borneol β-CDBao He solution, and with sweet mellow wine, it is an amount of to add water for injection, accent pH, and freeze-drying, promptly.

Preferred compound red sage root freezing-dried powder injection of the present invention, its preparation method is as follows:

B. above three flavors, red sage root boiling, collecting decoction, and concentrate, concentrate adds ethanol, leaves standstill, and filters, and filtrate concentrates, concentrate adds ethanol, leaves standstill, and filters, and filtrate concentrates, and adds water, and refrigeration filters, filtrate concentrates, and adds ethanol, leaves standstill, and filters, and filtrate concentrates, and drying gets Salvia root P.E; Pseudo-ginseng, boiling, collecting decoction concentrates, and adds ethanol, leaves standstill, filter, filtrate concentrates, and adds water, and refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-, add water, heating makes dissolving, adds activated charcoal and boils, and takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, inclusion filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boil, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add activated charcoal, charcoal is taken off in insulation, mix with borneol hydroxy propyl-Beta-CD inclusion solution filtrate cooling back, with sweet mellow wine, it is an amount of to add water for injection, transfers pH, freeze-drying, promptly.

B. above three flavors, red sage root section boiling 1～5 time, each 0.5～3 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 50～85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), add 2～14 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 50～90%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E; Get pseudo-ginseng, boiling 1～5 time, each 0.5～3 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 50～90%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 1～10 times of water gaging, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 50～85% ethanol elutions again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-100～1500g, add water in right amount, heating makes dissolving, adds 0.2～1% activated charcoal and boils 5～60 minutes, takes off charcoal, filtrate for later use.With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 1～12 hour filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 1～20 minute, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 50～90 ℃ of insulations of 0.05～0.2% activated charcoal 10～60 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 20～300g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, promptly.

Best compound red sage root freezing-dried powder injection of the present invention, its preparation method is as follows:

B. above three flavors, red sage root section boiling three times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is about 1.15 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 75%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.25 (50 ℃), add 8～10 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 80%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E; Get Radix Notoginseng powder and be broken into particle, boiling three times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 80%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adds 3～5 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 70% ethanol elution again, collect 70% ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-500g, add water to 1500ml, heating makes dissolving, adds 0.5% activated charcoal and boils 20 minutes, takes off charcoal, filtrate for later use.With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 6 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 10 minutes, put and be chilled to room temperature, refrigeration, filter, filtrate merges, and adds 80 ℃ of insulations of 0.1% activated charcoal 30 minutes, take off charcoal, filtrate is cooled to back below 50 ℃ mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 100g, add water for injection to 4000ml, transferring pH is 6.5, and degerming filters, can adds butyl rubber bung to cillin bottle, freeze-drying, gland, check, packing is promptly.

The freeze drying powder injection that medicine of the present invention is made, in every bottle of powder-injection, the weight of solid content is 0.78～0.83g.Average weight is the 0.8g/ bottle.

More than form when producing and to increase or to reduce according to corresponding ratio, as large-scale production can be unit with kilogram or with the ton, small-scale production can be unit with the gram also, and weight can increase or reduce, but the crude drug material weight proportion constant rate between each composition.

For a better understanding of the present invention, below learn the beneficial effect that the invention medicine is further set forth in test, test the effect that is intended to further specify medicine below by medicine effect of the present invention, but not limitation of the present invention.

Experimental example: the pharmacodynamics test of injection use compound red sage root freeze-dried powder

1 experiment material

1.1 medicine

Be subjected to the reagent thing: compound Danshen Root freeze-dried powder (sample before the freeze-drying): provide specification by Tianjin and this Bioisystech Co., Ltd: 300g/100ml/ bottle, lot number: 030307; Positive control medicine: Clary injection, Jiangsu Kang Yi pharmaceutical Co. Ltd product, product batch number: 030710-1.

1.2 reagent

Urethanes (urethane) is Shanghai San Pu chemical industry company limited product, lot number: 20030812; Yellow Jackets are Shanghai chemical reagents corporation product, lot number: F20020915; The pituitrin parenteral solution is by Shanghai first biochemical Pharma Inc. and biochemical-pharmaceutical factory, Shanghai Joint Production, lot number: 040301; Blue tetrazolium (UNI-CHEM Chemical Reogents), lot number: GD3010034; ADP and collagen, Sigma company product, final concentration are 3.5 μ mol/L.

1.3 instrument

BL-410 biological function experimental system is Sichuan Province Chengdu TME Technology Co., Ltd. product; The animal breathing apparatus, DH-150 type, Medical Instrument Factory affiliated to Zhejiang Univ production; It is Japanese photoelectricity company product that RM-6000 type four road physiology are led instrument more, and used plug-in unit is respectively floating ground formula bioelectric amplifier (AT-601G), carrier amplifier (AP-621G), blood pressure recorder (AP-611G), Cardiotacs (AT-601G); MF-1200 type Electromagnetic Flow is counted Japanese photoelectricity company product; Constant Temp. Oven is a Changsha medical apparatus and instruments factory product; HH-W600 digital display three is used constant temperature water tank, is Jintan City, Jiangsu Province high honour instrument Manufacturing Co., Ltd product; Laser Doppler flowmetry (PeriFluxSystem5000, PERIMED, USA); The LDZ5-2 hydro-extractor is Beijing Medical Centrifugal Machine Factory's product; Blood gas analyzer (USA Insyrumentation Laboratory, Gem-3000 type) and biochemical automated analysis instrument (USAVIROS-950 type); TYXN-91 type intelligence platelet aggregation instrument (Shanghai uncle bio tech ltd difficult to understand); System CA-530 Hemorheology instrument (U.S.).

1.4 dosage setting

With reference to clinical medicine dose, cause test such as rat heart muscle ischemic influence in conjunction with the chmice acute hypoxia endurance test, to clotting time of mice influence and to pituitrin and draw various animals administer dosage, as follows:

Rat low dosage: 0.30g/kg, middle dosage: 0.60g/kg, high dose: 1.20g/kg;

Dog low dosage: 0.09g/kg, middle dosage: 0.18g/kg, high dose: 0.36g/kg;

Mouse low dosage: 0.44g/kg, middle dosage: 0.88g/kg, high dose: 1.76g/kg;

Clary injection: dog: 0.53ml/Kg, rat: 1.8ml/Kg, mouse: 2.6ml/kg.

Drug dilution method, method of administration and volume: that gets same lot number respectively tests used medicine, with the physiological saline preparation, now prepares existing usefulness every day; Method of administration: intravenous injection or lumbar injection; Various units administration volume is respectively: mouse is the 0.15ml/10g body weight; Rat is 0.3ml/100g; Dog is the 3.0ml/kg body weight.

1.5 animal used as test

A cleaning level Kunming mouse, body weight 18～22g, male and female half and half, conformity certification number: 2002A055; A cleaning level SD rat, male and female half and half, body weight 180～230g, conformity certification number: 2003A070; New zealand white rabbit, body weight 2.0～2.5kg, male and female are not limit, conformity certification number: 2003A034; The hybrid dog, body weight 11.0～14.0kg provides by Zhongshan University's Experimental Animal Center.

The raising condition: after animal entered cleaning level Animal Lab., the every cage of mouse was put 10 in a suitable place to breed, and the every cage of rat is put 5 in a suitable place to breed, by special messenger's feeding and management.Animal housing's lighting every day 12 hours, heating ventilation and air-conditioning equipment is good, and room temperature is controlled at 25 ± 1 ℃, and relative humidity is 40～50%.The laboratory is sterilization regularly routinely.

1.6 statistical method

All (x ± s) expression relatively uses Student ' s t check to experimental result between the group of measurement data, relatively adopt paired t-test before and after the test of measurement data with average ± standard deviation.

2 experiment and results

2.2 compound Danshen Root is to the influence of acute myocardial ischemia dog cardiac muscle

2.2.1 experimental technique

30 of healthy hybrid dogs, body weight 11.0～14.0kg, male and female all have, be divided into dosage group (0.18g/kg), compound Danshen Root freeze-dried powder high dose group (0.36g/kg) and Clary injection group (0.53ml/kg) in sham operated rats, model group (physiological saline), compound Danshen Root freeze-dried powder low dose group (0.09g/kg), the compound Danshen Root freeze-dried powder at random, every group of 5 animals.3% yellow Jackets (30mg/kg) intravenous injection row general anesthesia, back of the body position is fixing.Standby from right jugular vein intubate as blood drawing.Tracheostomize inserts tracheal tube, connect the capable malleation artificial respiration of animal respirator (16 times/min).Right arm reclining is fixed, and opens chest in left IV intercostal, cuts off pericardium, exposes LADCA (LAD) threading standby (doing ligation), puts 9 electrodes and make thought-read adventitia electrograph and use on the pericardium of appropriate location down.Surgical injury finishes, treat that all observation indexs are stable after, record normal index and blood drawing.Ligation coronary artery left anterior descending branch (LAD) duplicates myocardial infarction and ischemia model, stablizes 30min record index, then respectively by each group dosage drip-feed administration (dripping off in the control 15min).The epicardial electrogram of each some when 30min, 60min, 120min after 30min and the administration before drawing blood respectively and writing down the ligation coronary artery, behind the ligation coronary artery.In the experimentation, the variation that continuous monitoring cardiogram II leads.

Trace epicardial electrogram and cardiogram with collection of BL-410 bio signal and analytical equipment.

2.2.2 observation index

1. to the influence of myocardial ischemia scope: 1. trace epicardial electrogram at each time point with BL-410 bio signal system respectively, cardiogram ischemic scope (N-ST) is raised the summation of counting that exceeds 2mv with ST section in whenever leading and is represented.Degree of myocardial ischemia (∑-ST) represent with the summation of rising of ST section or fall in whenever leading.2. to the mensuration of myocardial infarct size: behind 180min, take off heart and carry out the NBT of cardiac muscular tissue dyeing, measure the scope (infarcted region/left chamber heavy * 100%) of infarct with weight method.

2. sero-enzyme is measured: respectively at getting blood during 180min before pricking and after the administration, separation of serum is used automatic biochemical analyzer and is measured serum lactic dehydrogenase (SLDH) (LDH) and cretinephosphokinase (CPK).

2.2.3 experimental result

1. to the influence of myocardial ischemia dog myocardial ischemia scope

(1) to the influence of epicardial electrogram N-ST and ∑-ST

Following table 10 shows, after the model control group ligation after N-ST and ∑-ST ligation 30min 120min after the medication do not see significant change, injection gives the N-ST of compound Danshen Root freeze-dried powder and Clary injection group and ∑-ST all less than the ischemia model group, middle and high dosage is obvious especially, prompting compound Danshen Root freeze-dried powder can effectively dwindle the myocardial ischemia scope, alleviates the degree of myocardial ischemia.

Table 10 compound Danshen Root freeze-dried powder is to myocardial ischemia dog epicardial electrogram influence (n=5, x ± s)

Compare with model group, ^*P＜0.05, ^*P＜0.01

(2) to the influence of myocardial infarct size

Table 11 shows: each dosage group of compound Danshen Root freeze-dried powder can obviously reduce the myocardial ischemia scope, and high dose group is the most obvious, and middle dosage group is suitable with the positive control drug effect.

The influence of table 11 pair myocardial ischemia dog myocardial infarction scope (x ± s)

Compare with model group, ^*P＜0.05, ^*P＜0.01

2. to sero-enzyme (LDH, CK) influence of level

Following table is 12 results show, obviously do not change before and after sham operated rats dog myocardium enzyme LDH and the CK medication, and coronary ligation dog myocardium enzyme LDH and CK all significantly raise than sham operated rats or before performing the operation.Compound Danshen Root and Clary injection more all have the effect that reduces myocardial ischemia dog serum LDH and CPK level with model group.

The influence of table 12 couple myocardial ischemia dog LDH and CK (x ± s)

Compare with model group, ^*P＜0.05, ^*P＜0.01

3. to the influence of myocardial ischemia due to the pituitrin

3.1 experimental technique

70 of SD rats, body weight 180～230g divides 7 groups at random, and 10 every group, male and female half and half.With urethane 1200mg/Kg intraperitoneal injection of anesthesia.Back of the body position is fixing, gathers the lead electrocardiogram with analytical equipment record II with the BL-410 bio signal.Each organizes rat respectively from lumbar injection or irritate stomach and give following medicine: 1. model group (lumbar injection, ip.): physiological saline 3.0ml/kg; 2. compound Danshen Root freeze-dried powder low dosage (ip.) is organized: 0.30g/kg; 3. dosage (ip.) is organized in the compound Danshen Root freeze-dried powder: 0.60g/kg; 4. compound Danshen Root freeze-dried powder high dose (ip.) is organized: 1.20g/kg); 5. compound Danshen Root freeze-dried powder low dosage (is irritated stomach, po.) group: 0.60g/kg; 6. compound Danshen Root freeze-dried powder high dose (po.) is organized: 1.20g/kg; 7. Clary injection (ip.) is organized: 1.80ml/kg.Successive administration 3 days, once a day, behind the last administration 30min (1h after the administration of filling stomach group), each is organized rat and causes myocardial ischemia by sublingual vein injection of pituitrin 1u/kg respectively, observe and write down the before and after II lead electrocardiogram of 5s, 10s, 15s, 30s, 1min, 3min, 10min of every rat injection of pituitrin, measure the variation of each time point cardiogram index, each group result is carried out relatively reaching between group of administration front and back compare.

3.2 observation index

(1) J point displacement (mv) J point is the end of a period point of QRS ripple, and displacement is exactly the change that the position has taken place, and raises or forces down;

(2) T wave amplitude (mv)

(3) heart rate (inferior/min) behind the intravenous injection pituitrin, animal occurs the cardiogram ischemic immediately and changes, show as that the J point is raised or forced down, the T wave height is alarmmed or low flat, arrhythmia cordis such as premature ventricualr contraction, bradycardia, chamber conduction group stagnate etc., the variation of above several indexs before and after the pituitrin given in record.

3.3 experimental result

By table 13, table 14 and table 15 as seen, give pituitrin after, rat takes place obviously to change because of myocardial ischemia causes cardiogram, as J point raise, phenomenons such as the T wave height is alarmmed, arrhythmia cordis and decreased heart rate, change is the most remarkable during with 10s, 15s.Compare with model control group, the spoke degree of raising that basic, normal, high three the dosage groups of compound Danshen Root freeze-dried powder can obviously reduce the rat electrocardiogram J point due to the pituitrin (compares with model group, P＜0.05 or P＜0.01), the spoke degree of T ripple rising also obviously reduces (P＜0.05) than model group.Compound Danshen Root freeze-dried powder intravenously administrable effect is better than the effect of gastric infusion.

5. the compound Danshen Root freeze-dried powder is to the influence of blood platelet and thrombus

5.1 influence to extracorporeal platelet aggregation

Get blood from the rabbit arteria auricularis, get blood with the silication test tube, with 3.8% sodium citrate (1/10 blood volume) anti-freezing, centrifugal 10 minutes of 800r/min, sucking-off upper strata platelet rich plasma (PRP), centrifugal with 3000r/min again, take out platelet poor plasma (PPP).With PPP penetrability is transferred to 100, then with PRP zeroing, adds stirring rod, add physiological saline or compound Danshen Root freeze-dried powder (0.002,0.004,0.008g/ml) and Clary injection (0.05ml/ml), 37 ℃ of incubations.PRP is moved to the mensuration hole, add ADP (final concentration is 3.5 μ mol/L) or collagen (final concentration is 30 μ g/ml), observe the maximum aggregation extent of 5min, instrument is TYXN-91 type intelligence platelet aggregation instrument (Shanghai primary bio tech ltd difficult to understand).Calculate the platelet aggregation percent (Plateletaggregation, PAG) or suppress to assemble percent.

By following table 16,17 results as seen, the compound Danshen Root freeze-dried powder has the obvious suppression effect to ADP or collagen-induced rabbit platelet aggregation, along with the increase inhibiting effect of dosage strengthens gradually, promptly its inhibiting effect to ADP or collagen-induced rabbit platelet aggregation has tangible dose dependent.

The influence of the external rabbit platelet aggregation (PAG) that table 16 compound Danshen Root freeze-dried powder is induced ADP (x ± s)

Compare with the damping fluid control group, ^*P＜0.05, ^*P＜0.01

Table 17 compound Danshen Root freeze-dried powder is to the influence of collagen-induced external rabbit platelet aggregation (PAG) (x ± s)

Compare with the damping fluid control group, ^*P＜0.05, ^*P＜0.01

5.2 to the thrombotic influence of rat artery-vein bypass

50 of SD rats, be divided into 5 groups at random, every group 10, the anesthesia of 10% chloral hydrate 3.5ml/kg lumbar injection (ip) is fixing, respectively from sublingual vein give Clary injection (1.8ml/kg), compound Danshen Root freeze-dried powder high dose group (1.2g/kg), dosage group (0.6g/kg), low dosage (0.3g/kg), model group gives isometric physiological saline.30min after administration separates right carotid and left side vena jugularis externa.Put into No. 4 silk threads of a long 6cm in the polyethylene pipe stage casing, (50 μ l/ml) is full of polyethylene pipe with heparin-saline solution.After an end of polyethylene pipe inserted left vena jugularis externa, other end plastic tube injected heparin (50 μ l/m) 0.2ml anti-freezing, and then this end is inserted right carotid.Open artery clamp, allow blood in the right carotid inflow pipe, return the left side vena jugularis externa, behind the open blood 15min, middle clinopodium polycephalum, the thrombus that takes out rapidly in the polyethylene pipe is weighed on analytical balance, obtains thrombus inhibiting rate (%).

Thrombus inhibiting rate=[(control group wet weight of thrombus-administration group wet weight of thrombus) * 100%]/control group wet weight of thrombus

By following table 18 as seen, high, medium and low three dosage of compound Danshen Root freeze-dried powder all can obviously suppress the formation of rat arteriovenous shut thrombus, and at designed 0.3g/kg to 1.2g/kg, the increase effect of random quantity strengthens.

Table 18 compound Danshen Root freeze-dried powder to the total artery-vein of rat neck form the thrombus influence (x ± s, n=10)

Compare with model group ^*P＜0.05, ^*P＜0.01

6. the compound Danshen Root freeze-dried powder is to clotting time and hemorheological influence

6.1 the compound Danshen Root freeze-dried powder is to the influence of clotting time of mice

Method: 70 of mouse, body weight 18～22g, the male and female dual-purpose is divided into 7 groups at random by body weight and sex, and the administration volume is the 0.15ml/10g body weight.Give the intravenous injection normal saline solution respectively, 0.22g/kg, 0.44g/kg, the compound Danshen Root freeze-dried powder of 0.88g/kg, 1.76g/kg, 3.52g/kg, and 2.6ml/kg Clary injection.30min after the administration gets blood with glass capillary from eyeball, and slide method is measured the clotting time.

By following table 19 as seen, five dosage of compound Danshen Root freeze-dried powder 0.22～3.52g/kg all have the effect that prolongs clotting time of mice, and 0.22～1.76g/kg strengthens with the increase effect of agent, no longer increases greater than the 1.76g/kg effect.

Table 19 compound Danshen Root freeze-dried powder to the influence of clotting time of mice (x ± s, n=10)

6.2 the compound Danshen Root freeze-dried powder is to the influence of hemorheology of rat

Method: 50 of SD rats, be divided into 5 groups at random, every group 10, respectively from lumbar injection give Clary injection (1.8ml/kg), compound Danshen Root freeze-dried powder high dose group (1.2g/kg), dosage group (0.6g/kg), low dosage (0.3g/kg), control group gives isometric physiological saline.Every day 1 time, counted 7,30min after the last administration, use the etherization rat, get blood 1.8ml from rat arteria coeliaca, adding rapidly has in the mensuration pipe of 1: 9 sodium citrate solution 0.2ml, adopts and measures each whole blood viscosity, plasma viscosity (η p), blood relative viscosity (η r), erythrocyte aggregation index (AI), yield stress (CVS), blocks loose viscosity (CV) with hemorheology instrument (U.S.'s product).

The result: whole blood viscosity, plasma viscosity and the reduced viscosity of rat when following table 20 visible compound Danshen Root freeze-dried powder high dose group can obviously reduce different shear rate, reduce erythrocyte aggregation index, middle dosage group can obviously reduce height and cut whole blood viscosity.Prompting compound Danshen Root freeze-dried powder can improve the effect of Hemorheology function.

Table 20-1 compound Danshen Root freeze-dried powder to the influence of hemorheology of rat index (x ± s, n=10)

Compare with model group ^*P＜0.05, ^*P＜0.01

Table 20-2 compound Danshen Root freeze-dried powder to the influence of hemorheology of rat index (x ± s, n=10)

Compare with model group ^*P＜0.05, ^*P＜0.01

7. the compound Danshen Root freeze-dried powder is to the influence of mouse hypoxia-bearing capability

70 of mouse, male and female half and half are divided into 7 groups at random, 10 every group.The following medicine of intravenous injection respectively: 1. physiological saline 0.15ml/10g; 2. compound Danshen Root freeze-dried powder 0.22g/kg; 3. compound Danshen Root freeze-dried powder 0.44g/kg; 4. compound Danshen Root freeze-dried powder 0.88g/kg; 5. compound Danshen Root freeze-dried powder 1.76g/kg; 6. compound Danshen Root freeze-dried powder 3.52g/kg; 7. Clary injection 2.6ml/kg.The per injection capacity is 0.15ml/10g.Injected and distinguished lumbar injection isoprenaline hydrochloride (15mg/kg) again in 30 minutes, one by one mouse being put into capacity then is the anoxic bottle that 250ml, bottom are covered with the 5g soda lime, covers the bottle stopper that scribbles vaseline, causes the leakproofness anoxic.Write down the airtight bottleneck time, constantly observe small white mouse respiratory rate, the degree of depth, skin and lip change in color, cease breathing up to animal, write down the death time, from the lid bottle stopper to the breathless duration of animal during as the survival of mouse under the anoxia condition, time-to-live of medication group and control group relatively.

By table 21 as seen, each dosage group mouse time-to-live under the normobaric hypoxia condition of compound Danshen Root freeze-dried powder has prolonged 23.20% (P＞0.05), 35.57% (P＞0.05), 55.15% (P＜0.05), 76.29% (P＜0.01), 78.35% (P＜0.01) respectively than control group, prompting compound Danshen Root freeze-dried powder can improve the hypoxia-bearing capability of mouse, and is comparatively remarkable with 1.76g/kg.

Table 21 compound Danshen Root freeze-dried powder to the influence of hypoxia endurance time (x ± s, n=10)

Compare with model group ^*P＜0.05, ^*P＜0.01

Experiment showed, to dog intravenous injection compound Danshen Root freeze-dried powder 0.18g/kg and 0.36g/kg can increase the heart acting, obviously increase cardiac output and coronary blood flow; Dwindle cardiogram ischemic scope and the degree of myocardial ischemia of coronary ligation dog, the infringement that myocardial ischemia is caused has the protective effect of dose dependent.The compound Danshen Root freeze-dried powder can suppress platelet aggregation in addition, suppresses thrombus in vivo and forms, and prolongs the clotting time, improves hemorheology index and improves the mouse hypoxia-bearing capability.Showing that this medical instrument has significantly activates blood circulation and disperses blood clots and function of resisting myocardial ischemia, is the active drug of control myocardial ischemia disease, has important exploitation to be worth.This research prompting: the compound Danshen Root freeze-dried powder has the function that strengthens heart pump, increase coronary blood flow, resist myocardial ischemia-anoxemia, dwindle the functions such as cardiac damage due to the severe cardiac myocardial ischemia.

Embodiment

The present invention will be further described below in conjunction with embodiment, and following each embodiment only is used to the present invention is described and is not limitation of the present invention.

Embodiment 1:

A. get red sage root 1000g, pseudo-ginseng 850g, borneol 5g is standby;

B. the above red sage root, pseudo-ginseng process water respectively extract twice, and each 1 hour, merge extract, concentrate, get Salvia root P.E, Notogineng Extract; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Salvia root P.E, Notogineng Extract are added the injection water make dissolving in right amount, boil, put coldly, filter, filtrate is mixed with borneol β-CDBao He solution, and with sweet mellow wine, it is an amount of to add water for injection, accent pH, and freeze-drying is made 1000 bottles, promptly.

In the above-mentioned compound lyophilized powder, pseudo-ginseng is with the ginsenoside Rg1, the ginsenoside Rb1, and notoginsenoside R total amount meter, content is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D).

The linear gradient elution proportion of mobile phase changes program

This product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), ginsenoside Rb1, notoginsenoside R three's total amount meter, content is 18.75～56.25mg/g freeze-dried powder solid content.

Embodiment 2:

A. get red sage root 4000g, pseudo-ginseng 260g, borneol 50g is standby;

B. the above red sage root, pseudo-ginseng get Salvia root P.E, Notogineng Extract through using alcohol extract respectively; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), ginsenoside Rb1, notoginsenoside R three's total amount meter, content is 8.3～26.5mg/g freeze-dried powder solid content.

Embodiment 3:

A. get red sage root 1200g, pseudo-ginseng 300g, borneol 6g is standby;

B. the above red sage root, pseudo-ginseng are through carrying through water respectively, and 3 alcohol precipitations merge extract, concentrate, and get Salvia root P.E, Notogineng Extract; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 11.5～24.2mg/g freeze-dried powder solid content.

Embodiment 4

A. get red sage root 1500g, pseudo-ginseng 550g, borneol 20g is standby;

B. above red sage root water extracts twice, adds 8 times of water gagings extractions 3 hours for the first time, adds 6 times of water gagings for the second time and extracts 2 hours, merges extract, concentrates, and gets Salvia root P.E; Pseudo-ginseng alcohol extracting twice adds 8 times for the first time and measured 85% alcohol extracts 2 hours, adds 6 times of amount alcohol extracts 1 hour for the second time, merges extract, concentrates, and gets Notogineng Extract; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Salvia root P.E, Notogineng Extract or Salvia miltiorrhiza and Panax notoginseng extract are added injection water 500ml make dissolving, boiled 5 minutes, put coldly, filter, filtrate is mixed with borneol β-CDBao He solution, with sweet mellow wine 10g, add water for injection and supply 1000ml in right amount, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 18.5～42.2mg/g freeze-dried powder solid content.

Embodiment 5

A. get red sage root 1400g, pseudo-ginseng 350g, borneol 30g is standby;

B. above red sage root water extracts three times, adds 8 times of water gagings extractions 2 hours for the first time, adds 6 times of water gagings for the second time and extracts 1 hour, adds 4 times of water gagings for the third time and extracts 1 hour, merges extract, alcohol precipitation, and solution concentration gets Salvia root P.E; Pseudo-ginseng alcohol extracting three times adds 6 times of amount 80% alcohol extracts 2 hours for the first time, adds 6 times of amount 70% alcohol extracts 1 hour the second time, adds 4 times of amount 60% alcohol extracts 1 hour for the third time, merges extract, concentrates, and gets Notogineng Extract; An amount of dissolve with ethanol of borneol is used beta-cyclodextrin inclusion compound;

Salvia root P.E, Notogineng Extract or Salvia miltiorrhiza and Panax notoginseng extract are added injection water 300ml make dissolving, boiled 10 minutes, put coldly, filter, filtrate is mixed with borneol β-CDBao He solution, with sweet mellow wine 60g, add water for injection and supply 1000ml in right amount, transferring pH is 5～6.5, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 11.5～33.2mg/g freeze-dried powder solid content.

Embodiment 6

A. get red sage root 3500g, pseudo-ginseng 400g, borneol 8g is standby;

B. above three flavors, the red sage root adds 8 times of water gagings and decocts, collecting decoction, and concentrate, concentrate adds 60% ethanol, leaves standstill, and filters, and filtrate concentrates, concentrate adds 70% ethanol, leaves standstill, and filters, and filtrate concentrates, add water, filter, filtrate concentrates, add 80% ethanol, leave standstill, filter, filtrate concentrates, and drying gets Salvia root P.E;

Pseudo-ginseng adds 4 times of water gagings and decocts, and collecting decoction concentrates, and adds ethanol, leaves standstill, filter, filtrate concentrates, and adds water, and refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 85% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-, add water, heating makes dissolving, adds activated charcoal and boils, and takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, inclusion filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added injection water 200ml respectively make dissolving, boil, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add activated charcoal, charcoal is taken off in insulation, mix with borneol hydroxy propyl-Beta-CD inclusion solution filtrate cooling back, with sweet mellow wine, adds water for injection in right amount to 1000ml, transfer pH5～5.5, freeze-drying is made 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 19.5～31.2mg/g freeze-dried powder solid content.

Embodiment 7

A. get red sage root 2000g, pseudo-ginseng 450g, borneol 30g is standby;

B. above three flavors, the red sage root adds 6 times of water gagings and decocts 2 times, and each 1 hour, collecting decoction, and concentrate, concentrate adds 85% ethanol, leaves standstill, and filters, filtrate concentrates, and concentrate adds 80% ethanol, leaves standstill, and filters, filtrate concentrates, and adds water, filters, and filtrate concentrates, add 80% ethanol, leave standstill, filter, filtrate concentrates, and drying gets Salvia root P.E;

Pseudo-ginseng adds 7 times of water gagings and decocts, and collecting decoction concentrates, and adds 75% ethanol, leaves standstill, filter, filtrate concentrates, and adds water, and refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 75% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-, add water, heating makes dissolving, adds activated charcoal and boils, and takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, inclusion filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added injection water 250ml respectively make dissolving, boil, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add activated charcoal, charcoal is taken off in insulation, mix with borneol hydroxy propyl-Beta-CD inclusion solution filtrate cooling back, with sweet mellow wine, adds water for injection 1500ml, transfer pH4～5, freeze-drying is made 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 18.5～34.8mg/g freeze-dried powder solid content.

Embodiment 8

A. get red sage root 1800g, pseudo-ginseng 260g, borneol 10g is standby;

B. above three flavors, the red sage root adds 5 times of water gagings and decocts 2 times, and each 2 hours, collecting decoction, and concentrate, concentrate adds 75% ethanol, leaves standstill, and filters, filtrate concentrates, and concentrate adds 80% ethanol, leaves standstill, and filters, filtrate concentrates, and adds water, filters, and filtrate concentrates, add 80% ethanol, leave standstill, filter, filtrate concentrates, and drying gets Salvia root P.E;

Pseudo-ginseng adds 8 times of water gagings and decocts, and collecting decoction concentrates, and adds 85% ethanol, leaves standstill, filter, filtrate concentrates, and adds water, and refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 85% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-, add water, heating makes dissolving, adds activated charcoal and boils, and takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, inclusion filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added injection water 450ml respectively make dissolving, boil, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add activated charcoal, charcoal is taken off in insulation, mix with borneol hydroxy propyl-Beta-CD inclusion solution filtrate cooling back, with sweet mellow wine, adds water for injection 2000ml, transfer pH4～6, freeze-drying is made 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 9.5～25.4mg/g freeze-dried powder solid content.

Embodiment 9

A. get red sage root 3000g, pseudo-ginseng 360g, borneol 15 is standby;

B. above three flavors, red sage root section boiling 5 times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), add 4 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 90%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get pseudo-ginseng, boiling 5 times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 90%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 10 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 85% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-1500g, add water in right amount, heating makes dissolving, adds 1% activated charcoal and boils 60 minutes, takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 2 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 20 minutes, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 90 ℃ of insulations of 0.2% activated charcoal 60 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ and mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 300g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 1 method, this product contains pseudo-ginseng with ginsenoside Rg1 (C ₂₇H ₃₂O ₁₆), the total amount meter of ginsenoside Rb1, notoginsenoside R, content is 16.5～38.2mg/g freeze-dried powder solid content.

Embodiment 10

A. get red sage root 3000g, pseudo-ginseng 850g, borneol 35 is standby;

B. above three flavors, red sage root section boiling 3 times, each 2 hours, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 50%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), add 2 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 50%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get pseudo-ginseng, boiling 3 times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 50%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 2 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 50% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-100g, add water in right amount, heating makes dissolving, adds 0.2% activated charcoal and boils 5 minutes, takes off charcoal, filtrate for later use.With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 1～12 hour filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 1～20 minute, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 50 ℃ of insulations of 0.05% activated charcoal 10 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ and mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 20g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

In the above-mentioned compound lyophilized powder, pseudo-ginseng is with the ginsenoside Rg1, the ginsenoside Rb1, and notoginsenoside R content meter, content is measured according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia version in 2005 D).

1, instrument and reagent

Instrument high performance liquid chromatograph: Waters 600E pump, 717 automatic samplers, Waters PDA2996 diode array detector; Waters Empower chromatographic work station; Mettler AE240 (100,000/) balance (plum Teller-Tuo benefit Shanghai Instr Ltd.); High purity water distiller (Xinda, Jiangsu instrument plant) boils in SYZ-A quartz Asia.

The reagent ginsenoside Rg1 is available from Chinese medicine biological standardization institute (lot number 0703-200015 uses for containing to survey)

The ginsenoside Rb1 is available from Chinese biological drug inspection office (lot number 0704-200012 uses for containing to survey).

Notoginsenoside R is available from Chinese biological drug inspection office lot number (lot number 0745-200012 uses for containing to survey).

Acetonitrile is a chromatographically pure; Finished product sample and negative control product (providing) by our company.

2, the selection of chromatographic condition

Chromatographic column: Diamonsil C ₁₈, 5 μ m, 250 * 4.6mm (Di Ma company product); Mobile phase of acetonitrile and water linear gradient elution (as following table); Column temperature: 30 ℃; Flow velocity 1ml/min; Detect wavelength: 203nm (from Fig. 4-6 as can be seen, their absorption maximum is at 203nm, so select this wavelength).Under this chromatographic condition, the ginsenoside Rg1 separates well with other component in the sample, number of theoretical plate by ginsenoside Rg1's reference substance peak greater than 2000.(seeing Fig. 4 ~ 10).

The linear gradient elution proportion of mobile phase changes program

3, the preparation of need testing solution

Get this product content under the content uniformity item, mixing is got 0.5g, accurate claims surely, puts in the 10ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, and miillpore filter (0.45um) filters, promptly.

4, reappearance test

Precision takes by weighing 5 parts in sample, and (lot number: 030307), every part of about 0.5g presses need testing solution preparation method operation, and sample introduction is measured in accordance with the law, calculates content, asks relative standard deviation RSD.The result shows that this law is measured has good reappearance, the results are shown in Table 1.

Table 1 ginsenoside Rg1, ginsenoside Rb1, the test of notoginsenoside R reappearance

Embodiment 11

A. get red sage root 2469.85g, pseudo-ginseng 425.25g, borneol 25.25 is standby;

B. above three flavors, red sage root section boiling 2 times, each 1.5 hours, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 75%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 65%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), add 10 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 70%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get pseudo-ginseng, boiling 2 times, each 1.5 hours, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 70%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 6 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 65% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-200g, add water in right amount, heating makes dissolving, adds 0.5% activated charcoal and boils 30 minutes, takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 4 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 15 minutes, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 70 ℃ of insulations of 0.1% activated charcoal 20 minutes, take off charcoal, filtrate is cooled to back below 60 ℃ and mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 40g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 11 methods, this product contains the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 6.5～15.5mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁7.2～13.5mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁0.95～4.25mg/g freeze-dried powder solid content.

Embodiment 12

A. get red sage root 2266.2g, pseudo-ginseng 438.2g, natural borneol 20.6g is standby;

B. above three flavors, the red sage root adds 8 times of water gagings, second for the first time, add 6 times of water gagings for three times respectively, decocted 1 hour at every turn, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 65%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds ethanol and makes and contain the alcohol amount and reach 65%, leaves standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), adds 8 times of water gagings, refrigeration, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds ethanol and makes and contain the alcohol amount and reach 80%, leaves standstill, filter, filtrate is concentrated into thick paste, and drying gets Salvia root P.E;

Get pseudo-ginseng, add 6 times of water gagings and decoct 3 times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 60%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 6 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 65% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-150g, add water in right amount, heating makes dissolving, adds 0.4% activated charcoal and boils 20 minutes, takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 2 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 10 minutes, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 70 ℃ of insulations of 0.01% activated charcoal 10 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ and mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 80g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 11 methods, this product contains the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 7.5～13.5mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁7.9～15.5mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁1.36～4.05mg/g freeze-dried powder solid content.

Embodiment 13

A. get red sage root 2486.2g, pseudo-ginseng 486.2g, natural borneol 27.6g is standby;

B. above three flavors, the red sage root adds 8 times of water gagings, second for the first time, add 6 times of water gagings for three times respectively, decocted 1 hour at every turn, collecting decoction, and be concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 75%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adds ethanol and makes and contain the alcohol amount and reach 85%, leaves standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.25 (50 ℃), adds 8～10 times of water gagings, refrigeration, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adds ethanol and makes and contain the alcohol amount and reach 80%, leaves standstill, filter, filtrate is concentrated into thick paste, and drying gets Salvia root P.E;

Get Radix Notoginseng powder and be broken into particle, add 8 times of water gagings and decoct three times, each 1 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 80%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is about 1.15 (50 ℃), adds 3～5 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 70% ethanol elution again, collect 70% ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-500g, add water to 1500ml, heating makes dissolving, adds 0.5% activated charcoal and boils 20 minutes, takes off charcoal, filtrate for later use.With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 6 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 10 minutes, put and be chilled to room temperature, refrigeration, filter, filtrate merges, and adds 80 ℃ of insulations of 0.1% activated charcoal 30 minutes, take off charcoal, filtrate is cooled to back below 50 ℃ mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 100g, add water for injection to 4000ml, transferring pH is 6.5, and degerming filters, in can to the 1000 bottle cillin bottle, add butyl rubber bung, freeze-drying, gland, check, packing is promptly.

Measure content according to embodiment 11 methods, this product contains the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆), ginsenoside Rb ₁, Panax Notoginseng saponin R ₁See Table 2.

Table 2 ginsenoside Rg1, ginsenoside Rb1, the test of notoginsenoside R reappearance

Embodiment 14

A. get red sage root 2468.2g, pseudo-ginseng 468.2g, natural borneol 27.6g is standby;

B. above three flavors, red sage root section boiling 3 times, each 0.5 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃) adds ethanol and makes and contain the alcohol amount and reach 75%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.15～1.45 (50 ℃), add 4 times of water gagings, refrigeration filters, and filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get pseudo-ginseng, add 6 times of water gagings and decoct 3 times, each 0.5～3 hour, collecting decoction, and be concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), and add ethanol and make and contain the alcohol amount and reach 50～90%, leave standstill, filter, filtrate is concentrated into the medicinal extract that relative density is 1.05～1.35 (50 ℃), adds 5 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 75% ethanol elution again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-200g, add water in right amount, heating makes dissolving, adds 1% activated charcoal and boils 50 minutes, takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 8 hours filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 5 minutes, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 60 ℃ of insulations of 0.08% activated charcoal 25 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ and mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 150g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, make 1000 bottles, promptly.

Measure content according to embodiment 11 methods, this product contains the ginsenoside Rg ₁(C ₂₇H ₃₂O ₁₆) 8.012 ± 1.35mg/ bottle freeze-dried powder solid content, ginsenoside Rb ₁8.122 ± 1.45mg/ bottle freeze-dried powder solid content, Panax Notoginseng saponin R ₁2.049 ± 1.03mg/g freeze-dried powder solid content.

Claims

1. the content assaying method of a compound red sage root freezing-dried powder injection is characterized in that,

Described compound red sage root freezing-dried powder injection prepares by following method:

B. above three flavors, red sage root section boiling 1～5 time, each 0.5～3 hour, collecting decoction, and to be concentrated into relative density be 1.05～1.35 medicinal extract, 50 ℃ of mensuration, adding ethanol makes and contains the alcohol amount and reach 50～85%, leave standstill, filter, it is 1.05～1.35 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, adding ethanol makes and contains the alcohol amount and reach 50～85%, leave standstill, filter, it is 1.15～1.45 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, add 2～14 times of water gagings, refrigeration filters, and it is 1.05～1.35 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, adding ethanol makes and contains the alcohol amount and reach 50～90%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get pseudo-ginseng, boiling 1～5 time, each 0.5～3 hour, collecting decoction, and to be concentrated into relative density be 1.05～1.35 medicinal extract, 50 ℃ of mensuration, adds ethanol and make and contain the alcohol amount and reach 50～90%, leaves standstill, filter, it is 1.05～1.35 medicinal extract, 50 ℃ of mensuration that filtrate is concentrated into relative density, adds 1～10 times of water gaging, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 50～85% ethanol elutions again, collect ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract; Get HP-100～1500g, add water in right amount, heating makes dissolving, adds 0.2～1% activated charcoal and boils 5～60 minutes, takes off charcoal, filtrate for later use; With an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution, ultrasonic inclusion 1～12 hour filters, and gets borneol hydroxy propyl-Beta-CD inclusion solution;

Salvia root P.E and Notogineng Extract are added the injection water respectively make dissolving in right amount, boiled 1～20 minute, put and be chilled to room temperature, refrigeration filters, and filtrate merges, add 50～90 ℃ of insulations of 0.05～0.2% activated charcoal 10～60 minutes, take off charcoal, filtrate is cooled to back below 70 ℃ mixes with borneol hydroxy propyl-Beta-CD inclusion solution, with sweet mellow wine 20～300g, add water for injection to an amount of, transferring pH is 5～7, freeze-drying, promptly;

Adopt following method to measure ginsenoside Rg1 in the described freeze drying powder injection, ginsenoside Rb1, notoginsenoside R three's total amount, content assaying method is as follows:

Chromatographic condition and system suitability test are filling agent with octadecylsilane chemically bonded silica, are moving phase with the acetonitrile-water gradient elution; Gradient elution: 0min, 25% acetonitrile-75% water; 25min, 45% acetonitrile-55% water; 35min, 55% acetonitrile-45% water; Flow velocity 1.0ml/min; The detection wavelength is 203nm; Number of theoretical plate calculates by the ginsenoside Rg1 peak should be not less than 2000; The preparation of reference substance solution respectively precision to take by weighing ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R reference substance an amount of, add methyl alcohol respectively and make the solution that every 1ml contains 0.5mg, 0.5mg, 0.2mg, shake up, promptly;

This product content under the content uniformity item is got in the preparation of need testing solution, and mixing is got 0.5g, accurate claims surely, puts in the 10ml measuring bottle, is dissolved in water and is diluted to scale, shakes up, and the 0.45um miillpore filter filters, promptly;

Accurate respectively reference substance solution and each the 10 μ l of need testing solution of drawing of determination method inject liquid chromatograph, measure, and calculate, and ginsenoside Rg1, ginsenoside Rb1, notoginsenoside R three's total content is 6.25～56.25mg/g freeze-dried powder solid content.

2. content assaying method according to claim 1 is characterized in that: described compound red sage root freezing-dried powder injection prepares by following method:

B. above three flavors, red sage root section boiling three times, each 1 hour, collecting decoction, and to be concentrated into relative density be 1.15 medicinal extract, 50 ℃ of mensuration add ethanol and make and contain the alcohol amount and reach 75%, leave standstill, filter, it is 1.15 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, adding ethanol makes and contains the alcohol amount and reach 85%, leave standstill, filter, it is 1.25 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, add 8～10 times of water gagings, refrigeration filters, and it is 1.15 medicinal extract that filtrate is concentrated into relative density, 50 ℃ of mensuration, adding ethanol makes and contains the alcohol amount and reach 80%, leave standstill, filter, filtrate is concentrated into thick paste, drying gets Salvia root P.E;

Get Radix Notoginseng powder and be broken into particle, boiling three times, each 1 hour, collecting decoction, and to be concentrated into relative density be 1.15 medicinal extract, 50 ℃ of mensuration, adds ethanol and make and contain the alcohol amount and reach 80%, leaves standstill, filter, it is 1.15 medicinal extract, 50 ℃ of mensuration that filtrate is concentrated into relative density, adds 3～5 times of water gagings, refrigeration filters, and filtrate is by behind the macroporous adsorptive resins, wash with water earlier, use 70% ethanol elution again, collect 70% ethanol eluate, be concentrated into thick paste, drying gets Notogineng Extract;

Get HP-500g, add water to 1500ml, heating makes dissolving, adds 0.5% activated charcoal and boils 20 minutes, takes off charcoal, filtrate for later use, with an amount of dissolve with ethanol of borneol, slowly drop in the HP-aqueous solution ultrasonic inclusion 6 hours, filter, get borneol hydroxy propyl-Beta-CD inclusion solution;