CN1440773A - Prepn process of Qingkailing injection and injection powder and its quality control method - Google Patents
Prepn process of Qingkailing injection and injection powder and its quality control method Download PDFInfo
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Abstract
The present invention provides a improved preparation process of Qingkailing injection and powder for injection. The improved preparation process includes high speed centrifugal treatment in extracting cape jasmine, isatis root and honeysuckle, merging extractive liquid, ultrafiltering and other technological steps. It can produce Qingkailing injection product and powder product for injection with ever higher stability and ever longer effective period. The present invention also discloses the quality control method of baicalin, cholic acid, hyodeoxycholic acid, jasminoidin and chlorogenic acid in Qingkailing injection and powder for injectino as well as identification method of Qingkailing injection and power for injection.
Description
Technical field
The present invention relates to the preparation method of a kind of QINKAILING ZHUSHEYE or injection powder injection and the method for quality control of Qingkailing powder injection, specifically, be the improvement preparation method of a kind of QINKAILING ZHUSHEYE or injection powder injection and the method for quality control and the discrimination method of QINKAILING ZHUSHEYE or injection powder injection.
Background technology
QINGKAILING ZHUSHEJI is according to Chinese medical theory the Wu of the Qing Dynasty to be brought up in logical " the epidemic febrile disease bar is debated " " cow-bezoar bolus for resurrection " row side of tearing open analysisization year to come.Being widely used in patients such as hyperpyrexia, apoplexy clinically, is that numerous medical personnel generally acknowledge and habitual first-aid medicine.The research of relevant Qingkailing preparation aspect just began as far back as the seventies, had dosage forms such as " QINKAILING ZHUSHEYE ", " QINGKAILING KOUFUYE ", " QINGKAILING electuary " to come out and be applied to clinical in succession.Yet because injection inconvenience transportation, stability of Oral is poor, and the easy moisture absorption of electuary hardens, its clinical practice of disadvantages affect such as poor stability.In view of above-mentioned many reasons, people have developed lyophilized injectable powder on the basis of injection, and Freeze Drying Technique can make medicine keep original physicochemical property and physiologically active, and loss of effective components is few.In addition, the distinctive loose and porous structure of lyophilized formulations can make medicine be easy to again rehydration and recovers active, and the lyophilized formulations water content is low, easily steady in a long-term the preservation.Thereby both overcome the weak point of oral liquid, electuary, and overcome the shortcoming of injection inconvenience transportation again, and also kept former injection directly to enter body, rapid-action advantage has better curative effect for epidemic febrile disease, acute disease and hyperpyrexia disease.Nineteen ninety-five application such as Huang Cheng's steel has also obtained the patent of freeze-dried Qingkailing powder for injection, and this patent is to add excipient mannitol on the basis of QINKAILING ZHUSHEYE, aseptic filtration, and lyophilization is made.Preparation technology's patent application of the Qingkailing powder injection of Changcheng Pharmaceutical Factory, Zhangjiakou application was disclosed on October 1st, 1997, this technology is to be primary raw material with cholic acid 16.25g, Hyodeoxycholic Acid 18.75g, baicalin 25g, Concha Margaritifera powder 250g, Cornu Bubali powder 125g, Fructus Gardeniae 125g Radix Isatidis 1000g, Flos Lonicerae 100g, earlier the preparation Flos Lonicerae extractive solution; Isatis root extract; Gardenia extract; With cholic acid and hyodeoxycholic acid dissolving; The preparation Cornu Bubali hydrolysate; Preparation Concha Margaritifera powder hydrolyzed solution adds the Cornu Bubali powder hydrolyzed solution with the Concha Margaritifera powder hydrolyzed solution and prepares Cornu Bubali and Concha Margaritifera powder mixed liquor; Dissolve baicalin with water for injection; Mix isatis root extract and gardenia extract; Add cholic acid solution again; Add 95% ethanol to containing alcohol amount 70%, cold preservation 24 hours, filter, be concentrated into original volume 1/6-1/7, add Flos Lonicerae extractive solution again, with above-mentioned mixed liquor, Cornu Bubali and Concha Margaritifera mixed liquor and the merging of baicalin aqueous solution obtain total admixing medical solutions, add anhydrous sodium sulfite solution, benefit adds to the full amount of water for injection, and regulates pH to 6.5-7.0, add active carbon, water-bath was boiled 1 hour, put warm after-filtration, and filtrate cold preservation is placed and spent the night, sucking filtration, filtrate heating in water bath is again boiled half an hour, and cold preservation is spent the night again, through the microporous filter membrane sucking filtration, obtain the salmon pink medicinal liquid, lyophilization is made or spray drying is made.Make the Qingkailing powder injection that makes in this way, effect duration prolongs than aqueous injection, is easy to storing, and every index meets the pharmacopeia requirement.
However, we as can be seen the preparation method of above-mentioned Qingkailing powder injection be very loaded down with trivial details, be unfavorable for very much industrialized great production, and the quality of Qingkailing powder injection is still waiting further raising.Secondly, QINKAILING ZHUSHEYE or the powder ampoule agent for injection quality control to cholic acid, Hyodeoxycholic Acid, gardenoside and chlorogenic acid is not disclosed in the prior art.
Summary of the invention
For this reason, the present invention proposes the preparation method of a kind of QINKAILING ZHUSHEYE or powder ampoule agent for injection, this method is more suitable for suitability for industrialized production, and has improved the quality of QINKAILING ZHUSHEYE or powder ampoule agent for injection greatly.
Simultaneously, the present invention also provides the method for quality control of baicalin, cholic acid, Hyodeoxycholic Acid, gardenoside and chlorogenic acid in QINKAILING ZHUSHEYE or the injection powder injection.
The present invention also provides the discrimination method of QINKAILING ZHUSHEYE or powder ampoule agent for injection.
The inventive method has mainly been used ultracentrifugal technology in the leaching process of Fructus Gardeniae, Radix Isatidis and Flos Lonicerae, the impurity in effective like this extracting solution of removing these three kinds of Chinese crude drugs is for later simplification step is got ready.Be specifically
(1) preparation of gardenia extract:
Get Fructus Gardeniae, use the decocting in water alcohol deposition method, the filtrate high speed centrifugation makes gardenia extract, and is standby.
(2) preparation of isatis root extract:
Get the Radix Isatidis decoction pieces, use the decocting in water alcohol deposition method, the filtrate high speed centrifugation makes isatis root extract, and is standby.
(3) preparation of Flos Lonicerae extractive solution:
Extracting honeysuckle is used the decocting in water alcohol deposition method, and the filtrate high speed centrifugation makes Flos Lonicerae extractive solution, and is standby.
Centrifugal speed is in the 10000-20000 rev/min of scope in above-mentioned high speed centrifugation, and preferred centrifugal speed is 16000 rev/mins.
In addition, method of the present invention and in preparation Cornu Bubali and Concha Margaritifera powder mixture liquid step, added ultrafiltration step.Because Cornu Bubali contains compositions such as more protein and foreign preteins matter, can stimulate body to produce corresponding antibody, cause allergic reaction, clinical also have a more report.Compositions such as that the application of hyperfiltration technique has been removed effectively is protein-based, tannin have effectively been removed sensitinogen.
Specifically, the preparation of Cornu Bubali powder and Concha Margaritifera powder mixed hydrolysis liquid:
Get the Concha Margaritifera powder hydrolyzed solution, stir adding Cornu Bubali powder hydrolyzed solution down, regulate pH value to 4 with barium hydroxide solution, leave standstill, the leaching supernatant checks that barium ions is negative, and regulates pH value to 6～7 with sodium hydroxide solution, filter, filtrate is concentrated into 20 ℃, and to survey relative densities be 1.05 clear paste, puts cold, add ethanol, make to contain the alcohol amount and reach 70%, cold preservation, filter, decompression recycling ethanol is put cold to there not being the alcohol flavor, add ethanol, make to contain alcohol amount and reach 80%, cold preservation filters, decompression recycling ethanol, filter, with the concentrated solution ultrafiltration, the ultrafiltration liquid cooling is put, filter, promptly get Cornu Bubali and Concha Margaritifera powder mixed hydrolysis liquid.
Get the Concha Margaritifera powder hydrolyzed solution, stir adding Cornu Bubali powder hydrolyzed solution down, regulate pH value to 4 with barium hydroxide solution, leave standstill, the leaching supernatant is checked barium ions with 10% sulfuric acid solution, should be negative, regulate pH value to 6～7 with 10% sodium hydroxide solution, filter, filtrate is concentrated into the clear paste that relative density is 1.05 (20 ℃), puts cold, stir and add 95% ethanol down, make to contain alcohol amount and reach 70%, cold preservation 24 hours filters, decompression recycling ethanol is to there not being the alcohol flavor, put coldly, stir and to add ethanol down, make to contain the alcohol amount and reach 80%, cold preservation 24 hours, filter, decompression recycling ethanol filters, with the concentrated solution ultrafiltration, the ultrafiltration liquid cooling was put 24 hours, filtered, and promptly got Cornu Bubali and Concha Margaritifera powder mixed hydrolysis liquid.
Just because of adopted above-mentioned steps, make the total mixed liquor of preparation become easy, just, get Hyodeoxycholic Acid powder, baicalin powder, cholic acid powder respectively in addition the 10% sodium hydroxide solution water for injection of regulating pH value to 8 make dissolving in right amount, mixing; Add in the above-mentioned herbal extract, stir evenly, regulate pH value to 7, get mixed liquor, add to the full amount of water for injection, regulate pH value to 7; With the mixed liquor ultrafiltration, the filtrate lyophilization, promptly.
In the method for the invention, the aseptic filter membrane of 0.2-0.8 μ m, the aseptic filter membrane of preferred 0.22 μ m have been selected in ultrafiltration for use.
The method of quality control of baicalin wraps in and adds dissolve with methanol in the baicalin in QINKAILING ZHUSHEYE of the present invention or the powder ampoule agent for injection, filters, and gets baicalin solution, and is standby.High performance liquid chromatograph, with acetonitrile (or methanol)--(20～45: 80～55) be mobile phase, baicalin should be 90.0～110.0% of labelled amount to 0.3～0.6% phosphoric acid (or glacial acetic acid) in mensuration QINKAILING ZHUSHEYE or the powder ampoule agent for injection.
The method of quality control of cholic acid and Hyodeoxycholic Acid in QINKAILING ZHUSHEYE of the present invention or the powder ampoule agent for injection comprises cholic acid is added acetonitrile-phosphate buffer system dissolves, filters, and gets test liquid, and is standby; Use high performance liquid chromatograph, chromatographic column is C18 or C4 or C8 post, and mobile phase is with 40～85: 60～15: the methanol of 0.1～0.5 ratio or acetonitrile-water-glacial acetic acid or phosphatase assay contain cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount.
The method of quality control of gardenoside in QINKAILING ZHUSHEYE of the present invention or the powder ampoule agent for injection is included in and adds methanol-water dissolving in the gardenoside, filter, test liquid, standby; Use high performance liquid chromatograph, with 15～40: the methanol of 85～60 ratios or acetonitrile-water are mobile phase, and the product of measuring QINKAILING ZHUSHEYE or powder ampoule agent for injection contains gardenoside and must not be less than 1.52%.
The method of quality control of chlorogenic acid in QINKAILING ZHUSHEYE of the present invention or the powder ampoule agent for injection is included in and adds dissolve with methanol in the chlorogenic acid, filter, test liquid, standby; Use high performance liquid chromatograph, with 8～20: the acetonitrile of 92～80 ratios or methanol--0.2～0.6% phosphoric acid is mobile phase, and the product of measuring QINKAILING ZHUSHEYE or powder ampoule agent for injection contains chlorogenic acid and must not be less than 0.5%.
The quality standard that QINKAILING ZHUSHEYE that the inventive method is prepared or powder ampoule agent for injection use the said determination method to measure is that baicalin (C21H18011) should be 90.0～110.0% of labelled amount; Nitrogenous (N) should be 90.0～120.0% of labelled amount; Contain cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount; Contain gardenoside and must not be less than 1.52%, contain chlorogenic acid and must not be less than 0.5%.
The discrimination method of QINKAILING ZHUSHEYE of the present invention or powder ampoule agent for injection comprises that sample thief adds the alditol solution of 1-10ml new system, and with 1-10ml sulfuric acid solution mixing, heating in water bath, solution is gray purple; Sample thief adds dissolve with methanol simultaneously, filters, and filtrate concentrates, and gets test liquid; Get the Fructus Gardeniae medical material, add dissolve with methanol, filter, the spissated residue of filtrate adds methanol, gets Fructus Gardeniae medical material solution; Get gardenoside and add dissolve with methanol, get the reference substance solution test solution; According to thin layer chromatography, draw three kinds of solution, put in the pre-plate-making of GF254, with ethyl acetate-dehydrated alcohol-glacial acetic acid expansion of 8-12: 1-3: 0.5-1.5, spray is pressed from both sides blue aldehyde-concentrated sulfuric acid solution with perfume (or spice), in the test sample chromatograph with reference substance chromatograph relevant position on show the same color speckle arranged.
Use prepared QINKAILING ZHUSHEYE of the inventive method or injection powder injection, good stability, effect duration is long, and transportation is stored convenient, and every technical specification meets Pharmacopoeia of the People's Republic of China requirement.The using method of QINKAILING ZHUSHEYE of the present invention or injection powder injection is intramuscular injection: 2～4ml on the one.Patient with severe symptoms's intravenous injection 20～40ml uses with 10% glucose injection 200ml or normal saline 100ml dilution back.
Specifically preferred embodiment
Embodiment 1:
Cholic acid 32.5g Hyodeoxycholic Acid 37.5g baicalin 50g Concha Margaritifera powder 500g
The detailed process that Cornu Bubali powder 250g Fructus Gardeniae 250g Radix Isatidis 2000g Flos Lonicerae extract 50g prepares injectable powder goes on foot poly-as follows: 1, the extraction of medicine and hydrolysis process:
1. the preparation of gardenia extract: take by weighing Fructus Gardeniae by recipe quantity, add 6 times of water gagings and boiled 1 hour, filter, filtering residue adds 4 times of water gagings again, boil and carry half an hour, filter, filtrate merges, and heating is concentrated into the suitable volume of raw material weight, put and under agitation add dense ethanol after cold, make and contain alcohol amount and reach 60%, cold preservation 2～3 little days, sucking filtration, get clear filtrate, precipitation merges washing liquid and filtrate, decompression recycling ethanol with 60% cold washing with alcohol, 10000 rev/mins of ultracentrifugations, after the cold preservation 24 hours, filter to such an extent that clarify gardenia extract, cold preservation is placed for dosing and is used;
2. the preparation of isatis root extract: take by weighing the Radix Isatidis decoction pieces by recipe quantity, add 6～7 times of water gagings, boiled 1 hour, filter, filtering residue adds 4～5 times of water gagings again, boils half an hour, and the leaching extracting solution discards medicinal residues; When merging filtrate, heating make medicinal liquid be concentrated into the suitable volume of raw material weight, put coldly, under agitation add dense ethanol, reach 60% to containing the alcohol amount, cold preservation 2～3 days is filtered, decompression filtrate recycling ethanol, and concentrated solution cold preservation was placed 24 hours, filtered, and must clarify concentrated solution;
Get concentrated solution dropping ammonia under agitation, fully stir, make pH value transfer to 8.5～9.0, cold preservation was placed 24 hours, it is diffusing not allow ammonia wave, and with paper pulp cake sucking filtration, filtrate is heated in water-bath, removes ammonia to ammonia flavor and disappear under constantly stirring, to pH to 5.5～6.0 o'clock, cold preservation is placed, and uses for dosing;
3. the extraction of Flos Lonicerae extract: take by weighing Flos Lonicerae and put to boil and carry in the pot, add 15 times of water, little boil to boil carry 1 hour, filter, medicinal residues add 10 times of water gagings, boil and carry half an hour, filter, and discard medicinal residues; Merge filtrate twice, heating is concentrated into 1/8 of original volume～1/9 o'clock, and taking out with 20% lime cream adjusting pH is 12, the leaching precipitate;
Taking precipitate under agitation adds 2～3 times of amounts of precipitate, 95% ethanol in container, transfer pH=3～4 with 50% sulphuric acid, after fully stirring, filter, filtrate neutralizes with 40% sodium hydroxide, transfers pH=6.5～7, filter, filtrate recycling ethanol, 10000 rev/mins of ultracentrifugations, reconcentration is to the thick paste shape, put 80 ℃ of oven dry in the drying baker, get brown extract; Put in the exsiccator preserve standby;
4. take by weighing Cornu Bubali powder by recipe quantity, put in the beaker, add 8 times of amount 4N sodium hydroxide Ba (OH) 2 hot solution, be heated to and boil, constantly stir, be hydrolyzed 6～8 hours, treat that Cornu Bubali powder all is hydrolyzed into till the no brown granule; When putting temperature to 40～50 ℃, incline and get supernatant, filter, filtering residue is collected all solution with small amount of thermal washing 1～2 time, places standby;
5. press recipe quantity, take by weighing Concha Margaritifera powder, put in the flask, add 6～8 times of amount 4N sulfuric acid solutions, be heated to and boil, hydrolysis 6～8 hours, treat that Nacre transfers yellow mercury oxide (CaSO4 ↓) to by Lycoperdon polymorphum Vitt, stop heating, put temperature to 40～50 ℃, filter, filtering residue merges with small amount of thermal water washing 3 times, washing liquid and filtrate, puts cold back adularescent acicular crystal and separates out, refilter and remove, get light yellow transparent solution;
Barium sulfate impurity such as (BaSO4) is removed in neutralization: water intaking horn powder hydrolyzed solution, under agitation join in the Concha Margaritifera Usta hydrolysate, and after all Cornu Bubali hydrolysate adds,, add an amount of Ba (OH) 2 solids again as the mixed liquor peracid, the stirring and dissolving adjust pH is 4; Placement is sunk precipitation, ultrafiltration 10000 molecular weight, and leaching mixes hydrolyzed solution, wherein uses 10%H2SO4 fluid inspection Ba2+, should be negative, and reuse 10%NaOH liquid is transferred Ph=6～7, and sucking filtration gets faint yellow clear liquor;
Mixed hydrolysis liquid concentrates and alcohol is handled: clarifying mixed hydrolysis liquid is put in the evaporating dish, and heating evaporation concentrates, and during to 2～3 times of amounts of suitable raw material total amount, puts cold; Under agitation add dense ethanol, make to contain alcohol amount and reach 60%, cold preservation 24 hours is filtered, and decompression recycling ethanol does not have the alcohol flavor and ends near; Emitting cold preservation uses for dosing; 2, the preparation of injection:
(1) mixing of extraction medicinal liquid: water intaking Cornu Bovis seu Bubali and Concha Margaritifera mixed extract under agitation add isatis root extract; Add gardenia extract again; Mix homogeneously is regulated pH6～7, and sucking filtration gets clear and bright orange-red solution A;
(2) dissolving of taurine and hyocholic acid: it is an amount of to take 95% saturated ethanol of solid NaOH, stirs to be sprinkled into the taurine fine powder down gradually, transfers pH8.5～9.0, makes its whole dissolvings; Merge two sodium cholate solution, join in the A liquid;
(3) alcohol of admixing medical solutions is handled: admixing medical solutions such as water intaking Cornu Bovis seu Bubali, and add 95% ethanol and make and contain the alcohol amount and reach 70%, regulate pH6.5～7.0, cold preservation 24 hours is filtered, and decompression filtrate recycling ethanol is to there not being the alcohol flavor;
(4) baicalin and Flos Lonicerae extract are handled and dissolving:
Get an amount of water for injection, transfer about pH to 8, under agitation be sprinkled into the baicalin fine powder with 10%NaOH, make whole dissolvings after, transfer about pH to 7;
The extracting honeysuckle extract adds a small amount of water for injection, transfers with 95% ethanol to contain the alcohol amount and reach 80%, and cold preservation 24 hours, sucking filtration, precipitation 80% washing with alcohol merges washing liquid and filtrate, and decompression recycling ethanol is not to there being the alcohol flavor, continues concentrated golden yellow thick paste;
Dissolve purified Flos Lonicerae extractive solution with the solution that contains baicalin, incorporate in the admixing medical solutions, get total admixing medical solutions;
(5) processing of total admixing medical solutions: measure admixing medical solutions, add water for injection, regulate pH6.5～7.0 to 2000ml, add 0.2% active carbon, water-bath was boiled 1 hour, put warm after-filtration, and filtrate cold preservation is placed and spent the night, filter, filtrate is heated half an hour with boiling water bath, and cold preservation is spent the night again, with 0.2 μ m filter membrane ultrafiltration, get clear and bright Chinese red medicinal liquid, pH value should be 6.5～7.0; Be distributed into 2000 bottles, every bottle of 1ml, suitable crude drug 1.6g, lyophilization is promptly.
Embodiment 2:
Cholic acid 32.5g Hyodeoxycholic Acid 37.5g baicalin 50g Concha Margaritifera powder 500g
The detailed process that Cornu Bubali powder 250g Fructus Gardeniae 250g Radix Isatidis 2000g Flos Lonicerae extract 50g prepares injection goes on foot poly-as follows: 1, the extraction of medicine and hydrolysis process:
1. the preparation of gardenia extract: take by weighing Fructus Gardeniae by recipe quantity, add 6 times of water gagings and boiled 1 hour, filter, filtering residue adds 4 times of water gagings again, boil and carry half an hour, filter, filtrate merges, and heating is concentrated into the suitable volume of raw material weight, put and under agitation add dense ethanol after cold, make and contain alcohol amount and reach 60%, cold preservation 2～3 little days, sucking filtration, get clear filtrate, precipitation merges washing liquid and filtrate, decompression recycling ethanol with 60% cold washing with alcohol, 12000 rev/mins of ultracentrifugations, after the cold preservation 24 hours, filter to such an extent that clarify gardenia extract, cold preservation is placed for dosing and is used;
2. the preparation of isatis root extract: take by weighing the Radix Isatidis decoction pieces by recipe quantity, add 6～7 times of water gagings, boiled 1 hour, filter, filtering residue adds 4～5 times of water gagings again, boils half an hour, and the leaching extracting solution discards medicinal residues; When merging filtrate, heating make medicinal liquid be concentrated into the suitable volume of raw material weight, put coldly, under agitation add dense ethanol, reach 60% to containing the alcohol amount, cold preservation 2～3 days is filtered, decompression filtrate recycling ethanol, and concentrated solution cold preservation was placed 24 hours, filtered, and must clarify concentrated solution;
Get concentrated solution dropping ammonia under agitation, fully stir, make pH value transfer to 8.5～9.0, cold preservation was placed 24 hours, it is diffusing not allow ammonia wave, and with paper pulp cake sucking filtration, filtrate is heated in water-bath, removes ammonia to ammonia flavor and disappear under constantly stirring, to pH to 5.5～6.0 o'clock, cold preservation is placed, and uses for dosing;
3. the extraction of Flos Lonicerae extract: take by weighing Flos Lonicerae and put to boil and carry in the pot, add 15 times of water, little boil to boil carry 1 hour, filter, medicinal residues add 10 times of water gagings, boil and carry half an hour, filter, and discard medicinal residues; Merge filtrate twice, heating is concentrated into 1/8 of original volume～1/9 o'clock, and taking out with 20% lime cream adjusting pH is 12, the leaching precipitate;
Taking precipitate under agitation adds 2～3 times of amounts of precipitate, 95% ethanol in container, transfer pH=3～4 with 50% sulphuric acid, after fully stirring, filter, filtrate neutralizes with 40% sodium hydroxide, transfers pH=6.5～7, filter, filtrate recycling ethanol, 12000 rev/mins of ultracentrifugations, reconcentration is to the thick paste shape, put 80 ℃ of oven dry in the drying baker, get brown extract; Put in the exsiccator preserve standby;
4. take by weighing Cornu Bubali powder by recipe quantity, put in the beaker, add 8 times of amount 4N sodium hydroxide Ba (OH) 2 hot solution, be heated to and boil, constantly stir, be hydrolyzed 6～8 hours, treat that Cornu Bubali powder all is hydrolyzed into till the no brown granule; When putting temperature to 40～50 ℃, incline and get supernatant, filter, filtering residue is collected all solution with small amount of thermal washing 1～2 time, places standby;
5. press recipe quantity, take by weighing Concha Margaritifera powder, put in the flask, add 6～8 times of amount 4N sulfuric acid solutions, be heated to and boil, hydrolysis 6～8 hours, treat that Nacre transfers yellow mercury oxide (CaSO4 ↓) to by Lycoperdon polymorphum Vitt, stop heating, put temperature to 40～50 ℃, filter, filtering residue merges with small amount of thermal water washing 3 times, washing liquid and filtrate, puts cold back adularescent acicular crystal and separates out, refilter and remove, get light yellow transparent solution;
Barium sulfate impurity such as (BaSO4) is removed in neutralization: water intaking horn powder hydrolyzed solution, under agitation join in the Concha Margaritifera Usta hydrolysate, and after all Cornu Bubali hydrolysate adds,, add an amount of Ba (OH) 2 solids again as the mixed liquor peracid, the stirring and dissolving adjust pH is 4; Placement is sunk precipitation, ultrafiltration 10000 molecular weight, and leaching mixes hydrolyzed solution, wherein uses 10%H2SO4 fluid inspection Ba2+, should be negative, and reuse 10%NaOH liquid is transferred Ph=6～7, and sucking filtration gets faint yellow clear liquor;
Mixed hydrolysis liquid concentrates and alcohol is handled: clarifying mixed hydrolysis liquid is put in the evaporating dish, and heating evaporation concentrates, and during to 2～3 times of amounts of suitable raw material total amount, puts cold; Under agitation add dense ethanol, make to contain alcohol amount and reach 60%, cold preservation 24 hours is filtered, and decompression recycling ethanol does not have the alcohol flavor and ends near; Emitting cold preservation uses for dosing; 2, the preparation of injection:
(1) mixing of extraction medicinal liquid: water intaking Cornu Bovis seu Bubali and Concha Margaritifera mixed extract under agitation add isatis root extract; Add gardenia extract again; Mix homogeneously is regulated pH6～7, and sucking filtration gets clear and bright orange-red solution A;
(2) dissolving of taurine and hyocholic acid: it is an amount of to take 95% saturated ethanol of solid NaOH, stirs to be sprinkled into the taurine fine powder down gradually, transfers pH8.5～9.0, makes its whole dissolvings; Merge two sodium cholate solution, join in the A liquid;
(3) alcohol of admixing medical solutions is handled: admixing medical solutions such as water intaking Cornu Bovis seu Bubali, and add 95% ethanol and make and contain the alcohol amount and reach 70%, regulate pH6.5～7.0, cold preservation 24 hours is filtered, and decompression filtrate recycling ethanol is to there not being the alcohol flavor;
(4) baicalin and Flos Lonicerae extract are handled and dissolving:
Get an amount of water for injection, transfer about pH to 8, under agitation be sprinkled into the baicalin fine powder with 10%NaOH, make whole dissolvings after, transfer about pH to 7;
The extracting honeysuckle extract adds a small amount of water for injection, transfers with 95% ethanol to contain the alcohol amount and reach 80%, and cold preservation 24 hours, sucking filtration, precipitation 80% washing with alcohol merges washing liquid and filtrate, and decompression recycling ethanol is not to there being the alcohol flavor, continues concentrated golden yellow thick paste;
Dissolve purified Flos Lonicerae extractive solution with the solution that contains baicalin, incorporate in the admixing medical solutions, get total admixing medical solutions;
(5) processing of total admixing medical solutions: measure admixing medical solutions, add water for injection, regulate pH6.5～7.0 to 2000ml, add 0.2% active carbon, water-bath was boiled 1 hour, put warm after-filtration, and filtrate cold preservation is placed and spent the night, filter, filtrate is heated half an hour with boiling water bath, and cold preservation is spent the night again, with 0.5 μ m filter membrane ultrafiltration, get clear and bright Chinese red medicinal liquid, pH value should be 6.5～7.0; Be distributed into 2000 bottles, every bottle of 1ml, quite crude drug 1.6g is promptly.
Embodiment 3:
Cholic acid 32.5g Hyodeoxycholic Acid 37.5g baicalin 50g Concha Margaritifera powder 500g
The detailed process that Cornu Bubali powder 250g Fructus Gardeniae 250g Radix Isatidis 2000g Flos Lonicerae extract 50g prepares lyophilized injectable powder goes on foot poly-as follows: 1, the extraction of medicine and hydrolysis process:
1. the preparation of gardenia extract: take by weighing Fructus Gardeniae by recipe quantity, add 6 times of water gagings and boiled 1 hour, filter, filtering residue adds 4 times of water gagings again, boil and carry half an hour, filter, filtrate merges, and heating is concentrated into the suitable volume of raw material weight, put and under agitation add dense ethanol after cold, make and contain alcohol amount and reach 60%, cold preservation 2～3 little days, sucking filtration, get clear filtrate, precipitation merges washing liquid and filtrate, decompression recycling ethanol with 60% cold washing with alcohol, 16000 rev/mins of ultracentrifugations, after the cold preservation 24 hours, filter to such an extent that clarify gardenia extract, cold preservation is placed for dosing and is used;
2. the preparation of isatis root extract: take by weighing the Radix Isatidis decoction pieces by recipe quantity, add 6～7 times of water gagings, boiled 1 hour, filter, filtering residue adds 4～5 times of water gagings again, boils half an hour, and the leaching extracting solution discards medicinal residues; When merging filtrate, heating make medicinal liquid be concentrated into the suitable volume of raw material weight, put coldly, under agitation add dense ethanol, reach 60% to containing the alcohol amount, cold preservation 2～3 days is filtered, decompression filtrate recycling ethanol, and concentrated solution cold preservation was placed 24 hours, filtered, and must clarify concentrated solution;
Get concentrated solution dropping ammonia under agitation, fully stir, make pH value transfer to 8.5～9.0, cold preservation was placed 24 hours, it is diffusing not allow ammonia wave, and with paper pulp cake sucking filtration, filtrate is heated in water-bath, removes ammonia to ammonia flavor and disappear under constantly stirring, to pH to 5.5～6.0 o'clock, cold preservation is placed, and uses for dosing;
3. the extraction of Flos Lonicerae extract: take by weighing Flos Lonicerae and put to boil and carry in the pot, add 15 times of water, little boil to boil carry 1 hour, filter, medicinal residues add 10 times of water gagings, boil and carry half an hour, filter, and discard medicinal residues; Merge filtrate twice, heating is concentrated into 1/8 of original volume～1/9 o'clock, and taking out with 20% lime cream adjusting pH is 12, the leaching precipitate;
Taking precipitate under agitation adds 2～3 times of amounts of precipitate, 95% ethanol in container, transfer pH=3～4 with 50% sulphuric acid, after fully stirring, filter, filtrate neutralizes with 40% sodium hydroxide, transfers pH=6.5～7, filter, filtrate recycling ethanol, 16000 rev/mins of ultracentrifugations, reconcentration is to the thick paste shape, put 80 ℃ of oven dry in the drying baker, get brown extract; Put in the exsiccator preserve standby;
4. take by weighing Cornu Bubali powder by recipe quantity, put in the beaker, add 8 times of amount 4N sodium hydroxide Ba (OH) 2 hot solution, be heated to and boil, constantly stir, be hydrolyzed 6～8 hours, treat that Cornu Bubali powder all is hydrolyzed into till the no brown granule; When putting temperature to 40～50 ℃, incline and get supernatant, filter, filtering residue is collected all solution with small amount of thermal washing 1～2 time, places standby;
5. press recipe quantity, take by weighing Concha Margaritifera powder, put in the flask, add 6～8 times of amount 4N sulfuric acid solutions, be heated to and boil, hydrolysis 6～8 hours, treat that Nacre transfers yellow mercury oxide (CaSO4 ↓) to by Lycoperdon polymorphum Vitt, stop heating, put temperature to 40～50 ℃, filter, filtering residue merges with small amount of thermal water washing 3 times, washing liquid and filtrate, puts cold back adularescent acicular crystal and separates out, refilter and remove, get light yellow transparent solution;
Barium sulfate impurity such as (BaSO4) is removed in neutralization: water intaking horn powder hydrolyzed solution, under agitation join in the Concha Margaritifera Usta hydrolysate, and after all Cornu Bubali hydrolysate adds,, add an amount of Ba (OH) 2 solids again as the mixed liquor peracid, the stirring and dissolving adjust pH is 4; Placement is sunk precipitation, ultrafiltration 10000 molecular weight, and leaching mixes hydrolyzed solution, wherein uses 10%H2SO4 fluid inspection Ba2+, should be negative, and reuse 10%NaOH liquid is transferred Ph=6～7, and sucking filtration gets faint yellow clear liquor;
Mixed hydrolysis liquid concentrates and alcohol is handled: clarifying mixed hydrolysis liquid is put in the evaporating dish, and heating evaporation concentrates, and during to 2～3 times of amounts of suitable raw material total amount, puts cold; Under agitation add dense ethanol, make to contain alcohol amount and reach 60%, cold preservation 24 hours is filtered, and decompression recycling ethanol does not have the alcohol flavor and ends near; Emitting cold preservation uses for dosing; 2, the preparation of injection:
(1) mixing of extraction medicinal liquid: water intaking Cornu Bovis seu Bubali and extra large pearl oyster mixed extract under agitation add isatis root extract; Add gardenia extract again; Mix homogeneously is regulated pH6～7, and sucking filtration gets clear and bright orange-red solution A;
(2) dissolving of taurine and hyocholic acid: it is an amount of to take 95% saturated ethanol of solid NaOH, stirs to be sprinkled into the taurine fine powder down gradually, transfers pH8.5～9.0, makes its whole dissolvings; Merge two sodium cholate solution, join in the A liquid;
(3) alcohol of admixing medical solutions is handled: admixing medical solutions such as water intaking Cornu Bovis seu Bubali, and add 95% ethanol and make and contain the alcohol amount and reach 70%, regulate pH6.5～7.0, cold preservation 24 hours is filtered, and decompression filtrate recycling ethanol is to there not being the alcohol flavor;
(4) baicalin and Flos Lonicerae extract are handled and dissolving:
Get an amount of water for injection, transfer about pH to 8, under agitation be sprinkled into the baicalin fine powder with 10%NaOH, make whole dissolvings after, transfer about pH to 7;
The extracting honeysuckle extract adds a small amount of water for injection, transfers with 95% ethanol to contain the alcohol amount and reach 80%, and cold preservation 24 hours, sucking filtration, precipitation 80% washing with alcohol merges washing liquid and filtrate, and decompression recycling ethanol is not to there being the alcohol flavor, continues concentrated golden yellow thick paste;
Dissolve purified Flos Lonicerae extractive solution with the solution that contains baicalin, incorporate in the admixing medical solutions, get total admixing medical solutions;
(5) processing of total admixing medical solutions: measure admixing medical solutions, add water for injection, regulate pH6.5～7.0 to 2000ml, add 0.2% active carbon, water-bath was boiled 1 hour, put warm after-filtration, and filtrate cold preservation is placed and spent the night, filter, filtrate is heated half an hour with boiling water bath, and cold preservation is spent the night again, with 0.8 μ m filter membrane ultrafiltration, get clear and bright Chinese red medicinal liquid, pH value should be 6.5～7.0; Be distributed into 2000 bottles, every bottle of 1ml, suitable crude drug 1.6g, lyophilization is promptly.
Embodiment 4: the method for quality control of QINKAILING ZHUSHEYE or powder ampoule agent for injection
Differentiate: a. gets 1 bottle of this product and adds the dissolving of 5ml water for injection, gets the furfural that 0.5ml adds new preparation
Solution (1 → 10ml) 1ml, sulfuric acid solution (get sulphuric acid 50ml, add water 65ml and mix) 10ml,
Mixing, heating is 10 minutes in 70 ℃ of water-baths, and solution shows gray purple;
B. get this product and add the 5ml dissolve with methanol for 1 bottle, filter, filtrate decompression is concentrated into 1ml, as confession
Test sample solution takes by weighing Fructus Gardeniae medical material 0.5g, adds methanol 10ml lixiviate and spends the night, and filters, and the residue after filtrate concentrates adds that methanol is molten to get Fructus Gardeniae medical material solution to 2ml; Taking by weighing gardenoside 2mg again, to add methanol molten to 1ml product solution in contrast; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica GF254 precoated plate, launch with ethyl acetate-dehydrated alcohol of 10: 2: 1-glacial acetic acid, dry, spray is with the blue aldehyde-concentrated sulfuric acid solution of 1% fragrant folder, 105 ℃ of bakings 5 minutes, in the test sample chromatograph with the corresponding position of reference substance chromatograph on show the same color speckle arranged;
Assay:
A. baicalin,
Get about Qingkailing powder injection sample 3mg, the accurate title, decide, and puts in the 5ml volumetric flask, and it is molten to scale to add methanol, and the aperture is 0.45 μ m filtering with microporous membrane, gets subsequent filtrate as test solution; Other gets the baicalin reference substance, and accurate the title decides, and adds dissolve with methanol and makes the solution that every 1ml contains 50 μ g, in contrast product solution; Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia nineteen ninety-five version D); Respectively accurate each the 10 μ l of baicalin reference substance solution and need testing solution that draw inject high performance liquid chromatograph, and mobile phase was with 28: 72 acetonitrile-0.025M phosphoric acid, and the detection wavelength is 276nm, mensuration peak area integrated value, with external standard method calculating promptly; This product contains baicalin (C21H18011) and should be 90.0～110.0% of labelled amount;
B. cholic acid, Hyodeoxycholic Acid:
The reference substance solution preparation, precision takes by weighing cholic acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0; Precision takes by weighing Hyodeoxycholic Acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0; The test liquid preparation: precision takes by weighing Qingkailing powder injection sample 20mg and puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively each the 10 μ l of cholic acid, Hyodeoxycholic Acid reference substance solution and test liquid that draw, inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase was with 35: 65 acetonitrile-0.025M phosphate pH3.0, flow velocity 1.0ml/min, the detection wavelength is 205nm, measures cholic acid, Hyodeoxycholic Acid peak area integrated value, calculates with external standard method; This product contains cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount;
C. gardenoside:
The reference substance solution preparation, precision takes by weighing gardenoside standard substance 1mg, puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters; The test liquid preparation: precision takes by weighing Qingkailing powder injection sample 5mg and puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively gardenoside reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 30: 70 methanol-waters, flow velocity 1.0ml/min, and the detection wavelength is 237nm, measure gardenoside peak area integrated value, calculate with external standard method; This product contains gardenoside must not be less than 1.52%;
D. chlorogenic acid:
The reference substance solution preparation, precision takes by weighing chlorogenic acid 1mg and puts in the 10ml volumetric flask, and it is molten to scale to add methanol; The test liquid preparation: precision takes by weighing sample 20mg, puts in the 2ml volumetric flask, and it is molten to scale to add methanol, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively chlorogenic acid reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 10: 90 acetonitrile-0.025M phosphate, flow velocity 1.0ml/min, and the detection wavelength is 326nm, measure chlorogenic acid peak area integrated value, calculate with external standard method; This product contains chlorogenic acid must not be less than 0.5%;
E. nitrogen content,
Precision takes by weighing this product 2mg, the photograph N2 method (" an appendix IX of Chinese pharmacopoeia nineteen ninety-five version L second method) measure, that is, this product nitrogenous (N) should be 90.0～120.0% of labelled amount; Embodiment 5: the discrimination method of injection of the present invention or injection powder injection
Differentiate: a. gets 1 bottle of Qingkailing powder injection and adds dissolving of 5ml water for injection or QINKAILING ZHUSHEYE 5ml, (1 → 10ml) 1ml, sulfuric acid solution (are got sulphuric acid 50ml to get the furfural solution that 0.5ml adds new preparation, adding water 65ml mixes) 10ml, mixing, heating is 10 minutes in 70 ℃ of water-baths, and solution shows gray purple;
B. get 1 bottle of power 5ml of Qingkailing powder injection dissolve with methanol, filter, filtrate decompression is concentrated into 1ml, as need testing solution, takes by weighing Fructus Gardeniae medical material 0.5g, adds methanol 10ml lixiviate and spends the night, and filters, and the residue after filtrate concentrates adds that methanol is molten to get Fructus Gardeniae medical material solution to 2ml; Taking by weighing gardenoside 2mg again, to add methanol molten to 1ml product solution in contrast; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica GF254 precoated plate, launch with ethyl acetate-dehydrated alcohol of 10: 2: 1-glacial acetic acid, dry, spray is with the blue aldehyde-concentrated sulfuric acid solution of 1% fragrant folder, 105 ℃ of bakings 5 minutes, in the test sample chromatograph with the corresponding position of reference substance chromatograph on show the same color speckle arranged;
Assay:
A. baicalin,
Get about Qingkailing powder injection 3mg, the accurate title, decide, and puts in the 5ml volumetric flask, and it is molten to scale to add methanol, and the aperture is 0.45 μ m filtering with microporous membrane, gets subsequent filtrate as test solution; Other gets the baicalin reference substance, and accurate the title decides, and adds dissolve with methanol and makes the solution that every 1ml contains 50 μ g, in contrast product solution; Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia nineteen ninety-five version D); Respectively accurate each the 10 μ l of baicalin reference substance solution and need testing solution that draw inject high performance liquid chromatograph, and mobile phase was with 30: 70 acetonitrile-0.025M phosphoric acid, and the detection wavelength is 276nm, mensuration peak area integrated value, with external standard method calculating promptly; This product contains baicalin (C21H18O11) and should be 90.0～110.0% of labelled amount;
B. cholic acid, Hyodeoxycholic Acid:
The reference substance solution preparation, precision takes by weighing cholic acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0; Precision takes by weighing Hyodeoxycholic Acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH 3.0; The test liquid preparation: precision takes by weighing this product 20mg and puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively each the 10 μ l of cholic acid, Hyodeoxycholic Acid reference substance solution and test liquid that draw, inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase was with 38: 62 acetonitrile-0.025 M phosphate pH3.0, flow velocity 1.0ml/min, the detection wavelength is 205nm, measures cholic acid, Hyodeoxycholic Acid peak area integrated value, calculates with external standard method; This product contains cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount;
C. gardenoside:
The reference substance solution preparation, precision takes by weighing gardenoside standard substance 1mg, puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters; The test liquid preparation: precision takes by weighing Qingkailing powder injection 5mg and puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively gardenoside reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 35: 65 methanol-waters, flow velocity 1.0ml/min, and the detection wavelength is 237nm, measure gardenoside peak area integrated value, calculate with external standard method; This product contains gardenoside must not be less than 1.52%;
D. chlorogenic acid:
The reference substance solution preparation, precision takes by weighing chlorogenic acid 1mg and puts in the 10ml volumetric flask, and it is molten to scale to add methanol; The test liquid preparation: precision takes by weighing Qingkailing powder injection sample 20mg, puts in the 2ml volumetric flask, and it is molten to scale to add methanol, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively chlorogenic acid reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 15: 85 acetonitrile-0.025 M phosphate, flow velocity 1.0ml/min, and the detection wavelength is 326nm, measure chlorogenic acid peak area integrated value, calculate with external standard method; This product contains chlorogenic acid must not be less than 0.5%;
E. nitrogen content,
Precision takes by weighing Qingkailing powder injection sample 2mg, the photograph N2 method (" an appendix IX of Chinese pharmacopoeia nineteen ninety-five version L second method) measure, that is, this product nitrogenous (N) should be 90.0～120.0% of labelled amount; Embodiment 6: the method for quality control of injection of the present invention or powder ampoule agent for injection
Differentiate: a. gets 1 bottle of Qingkailing powder injection and adds the dissolving of 5ml water for injection or get QINKAILING ZHUSHEYE 5ml, (1 → 10ml) 1ml, sulfuric acid solution (are got sulphuric acid 50ml to get the furfural solution that 0.5ml adds new preparation, adding water 65ml mixes) 10ml, mixing, heating is 10 minutes in 70 ℃ of water-baths, and solution shows gray purple;
B. get this product and add the 5ml dissolve with methanol for 1 bottle, filter, filtrate decompression is concentrated into 1ml, as need testing solution, takes by weighing Fructus Gardeniae medical material 0.5g, adds methanol 10ml lixiviate and spends the night, and filters, and the residue after filtrate concentrates adds that methanol is molten to get Fructus Gardeniae medical material solution to 2ml; Taking by weighing gardenoside 2mg again, to add methanol molten to 1ml product solution in contrast; Test according to thin layer chromatography (an appendix VI of Chinese Pharmacopoeia B), draw each 5 μ l of above-mentioned three kinds of solution, put respectively on same silica GF254 precoated plate, launch with ethyl acetate-dehydrated alcohol of 10: 2: 1-glacial acetic acid, dry, spray is with the blue aldehyde-concentrated sulfuric acid solution of 1% fragrant folder, 105 ℃ of bakings 5 minutes, in the test sample chromatograph with the corresponding position of reference substance chromatograph on show the same color speckle arranged;
Assay:
The a baicalin,
Get about Qingkailing powder injection sample 3mg, the accurate title, decide, and puts in the 5ml volumetric flask, and it is molten to scale to add methanol, and the aperture is 0.45 μ m filtering with microporous membrane, gets subsequent filtrate as test solution; Other gets the baicalin reference substance, and accurate the title decides, and adds dissolve with methanol and makes the solution that every 1ml contains 50 μ g, in contrast product solution; Measure according to high performance liquid chromatography (" an appendix VI of Chinese pharmacopoeia nineteen ninety-five version D); Respectively accurate each the 10 μ l of baicalin reference substance solution and need testing solution that draw inject high performance liquid chromatograph, and mobile phase was with 40: 60 acetonitrile-0.025M phosphoric acid, and the detection wavelength is 276nm, mensuration peak area integrated value, with external standard method calculating promptly; This product contains baicalin (C21H18O11) and should be 90.0～110.0% of labelled amount;
B. cholic acid, Hyodeoxycholic Acid:
The reference substance solution preparation, precision takes by weighing cholic acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0; Precision takes by weighing Hyodeoxycholic Acid 2mg, puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0; The test liquid preparation: precision takes by weighing this product 20mg and puts in the 2ml volumetric flask, and it is molten to scale to add 35: 65 acetonitrile-0.025M phosphate buffer pH3.0, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively each the 10 μ l of cholic acid, Hyodeoxycholic Acid reference substance solution and test liquid that draw, inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase was with 30: 70 acetonitrile-0.025 M phosphate pH3.0, flow velocity 1.0ml/min, the detection wavelength is 205nm, measures cholic acid, Hyodeoxycholic Acid peak area integrated value, calculates with external standard method; This product contains cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount;
C. gardenoside:
The reference substance solution preparation, precision takes by weighing gardenoside standard substance 1mg, puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters; The test liquid preparation: precision takes by weighing Qingkailing powder injection sample 5mg and puts in the 2ml volumetric flask, and it is molten to scale to add 30: 70 methanol-waters, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively gardenoside reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 20: 80 methanol-waters, flow velocity 1.0ml/min, and the detection wavelength is 237nm, measure gardenoside peak area integrated value, calculate with external standard method; This product contains gardenoside must not be less than 1.52%;
D. chlorogenic acid:
The reference substance solution preparation, precision takes by weighing chlorogenic acid 1mg and puts in the 10ml volumetric flask, and it is molten to scale to add methanol; The test liquid preparation: precision takes by weighing sample 20mg, puts in the 2ml volumetric flask, and it is molten to scale to add methanol, and filtering with microporous membrane gets test liquid; " an appendix VI of Chinese pharmacopoeia D measures according to high performance liquid chromatography; Accurate respectively chlorogenic acid reference substance solution and each 10 μ l of test liquid of drawing inject high performance liquid chromatograph, chromatographic column C18ODS post, mobile phase is with 18: 82 acetonitrile-0.025 M phosphate, flow velocity 1.0ml/min, and the detection wavelength is 326nm, measure chlorogenic acid peak area integrated value, calculate with external standard method; This product contains chlorogenic acid must not be less than 0.5%;
E. nitrogen content,
Precision takes by weighing Qingkailing powder injection sample 2mg, the photograph N2 method (" an appendix IX of Chinese pharmacopoeia nineteen ninety-five version L second method) measure, that is, this product nitrogenous (N) should be 90.0～120.0% of labelled amount.
Claims (10)
1. the preparation method of QINKAILING ZHUSHEYE or powder ampoule agent for injection, it is with cholic acid 16.25g, Hyodeoxycholic Acid 18.75g, baicalin 25g, Concha Margaritifera powder 250g, Cornu Bubali powder 125g, Fructus Gardeniae 125g Radix Isatidis 1000g, Flos Lonicerae 100g is a primary raw material, with Fructus Gardeniae, Radix Isatidis and Flos Lonicerae are extracted respectively and make extracting solution, then Cornu Bubali is made hydrolyzed solution, Concha Margaritifera powder is made hydrolyzed solution, it is characterized in that preparing Fructus Gardeniae respectively, in Radix Isatidis and the Flos Lonicerae extractive solution extracting solution has all been carried out high speed centrifugation, get the Concha Margaritifera powder hydrolyzed solution then, stir and add the Cornu Bubali powder hydrolyzed solution down, regulate pH value to 4 with barium hydroxide solution, leave standstill, the leaching supernatant checks that barium ions is negative, and regulates pH value to 6～7 with sodium hydroxide solution, filter, filtrate is concentrated into 20 ℃, and to survey relative densities be 1.05 clear paste, puts coldly, adds ethanol, make and contain alcohol amount and reach 70%, cold preservation filters, and decompression recycling ethanol is to there not being the alcohol flavor, put cold, add ethanol, make to contain the alcohol amount and reach 80%, cold preservation, filter, decompression recycling ethanol filters, with the concentrated solution ultrafiltration, the ultrafiltration liquid cooling is put, filter, promptly get Cornu Bubali and Concha Margaritifera powder mixed hydrolysis liquid, standby; Get Hyodeoxycholic Acid powder, baicalin powder, cholic acid powder respectively in addition the sodium hydroxide solution water for injection of regulating pH value to 8 make dissolving, mixing; Add in the above-mentioned herbal extract, stir evenly, regulate pH value to 7, get mixed liquor, add to the full amount of water for injection, regulate pH value to 7; With the mixed liquor ultrafiltration, filtrate must injection, and perhaps lyophilization or spray drying promptly get powder ampoule agent for injection.
2. in accordance with the method for claim 1, wherein ultracentrifugal speed is in the 10000-20000 rev/min of scope.
3. according to the described method of claim 2, wherein ultracentrifugal speed is 16000 rev/mins.
4. according to any one described method among the claim 1-3, wherein ultrafiltration is with the aseptic filter membrane of 0.2-0.8 μ m.
5. in accordance with the method for claim 4, wherein ultrafiltration with the aseptic filter membrane of 0.22 μ m.
6. the method for quality control of baicalin in QINKAILING ZHUSHEYE or the powder ampoule agent for injection, it comprises baicalin is added dissolve with methanol, filter, baicalin solution, standby; Use high performance liquid chromatograph, with 20～45: the acetonitrile of 80～55 ratios or methanol-0.3～0.6% phosphoric acid or glacial acetic acid are mobile phase, measure QINKAILING ZHUSHEYE or powder ampoule agent for injection, wherein contain baicalin and should be 90.0～110.0% of labelled amount.
7. the method for quality control of cholic acid and Hyodeoxycholic Acid in QINKAILING ZHUSHEYE or the powder ampoule agent for injection, it comprises cholic acid is added acetonitrile-phosphate buffer system dissolves, filter, test liquid, standby; Use high performance liquid chromatograph, chromatographic column is C18 or C4 or C8 post, mobile phase is with 40～85: 60～15: the methanol of 0.1～0.5 ratio or acetonitrile-water-glacial acetic acid or phosphatase assay QINKAILING ZHUSHEYE or powder ampoule agent for injection contain cholic acid, Hyodeoxycholic Acid should be 90.0%～110.0% of labelled amount.
8. the method for quality control of gardenoside in QINKAILING ZHUSHEYE or the powder ampoule agent for injection, it is included in and adds the methanol-water dissolving in the gardenoside, filter, test liquid, standby; Use high performance liquid chromatograph, with 15～40: the methanol of 85～60 ratios or acetonitrile-water are mobile phase, measure the product of QINKAILING ZHUSHEYE or powder ampoule agent for injection, contain gardenoside and must not be less than 1.52%.
9. the method for quality control of chlorogenic acid in QINKAILING ZHUSHEYE or the powder ampoule agent for injection, it is included in and adds dissolve with methanol in the chlorogenic acid, filter, test liquid, standby; Use high performance liquid chromatograph, with 8～20: the acetonitrile of 92～80 ratios or methanol-0.2～0.6% phosphoric acid is mobile phase, measures the product of QINKAILING ZHUSHEYE or powder ampoule agent for injection, contains chlorogenic acid and must not be less than 0.5%.
10. the discrimination method of QINKAILING ZHUSHEYE or powder ampoule agent for injection, it comprises gets the alditol solution that QINKAILING ZHUSHEYE or powder ampoule agent for injection sample add the 1-10ml new system, and with 1-10ml sulfuric acid solution mixing, heating in water bath, solution is gray purple; Get QINKAILING ZHUSHEYE or powder ampoule agent for injection sample simultaneously and filter, the spissated residue of filtrate adds methanol, gets Fructus Gardeniae medical material solution; Get gardenoside and add dissolve with methanol, get the reference substance solution test solution; According to thin layer chromatography, draw three kinds of solution, point launches spray with ethyl acetate-dehydrated alcohol-glacial acetic acid of 8-12: 1-3: 0.5-1.5 and presss from both sides blue aldehyde-concentrated sulfuric acid solution with perfume (or spice) in the pre-plate-making of GF254, in the test sample chromatograph with reference substance chromatograph relevant position on show the same color speckle arranged.
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| CN1299717C (en) * | 2004-08-06 | 2007-02-14 | 北京中医药大学 | Pharmaceutical composition for treating cerebral apoplexy and its preparation method |
| CN100376260C (en) * | 2005-08-29 | 2008-03-26 | 贵州益佰制药股份有限公司 | Method for preparing Qingkailing preparation |
| CN1739655B (en) * | 2005-09-14 | 2010-04-14 | 贵州益佰制药股份有限公司 | Quality control method of Qingkailing injection preparation |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1299717C (en) * | 2004-08-06 | 2007-02-14 | 北京中医药大学 | Pharmaceutical composition for treating cerebral apoplexy and its preparation method |
| CN100376260C (en) * | 2005-08-29 | 2008-03-26 | 贵州益佰制药股份有限公司 | Method for preparing Qingkailing preparation |
| CN1739655B (en) * | 2005-09-14 | 2010-04-14 | 贵州益佰制药股份有限公司 | Quality control method of Qingkailing injection preparation |
| CN1813812B (en) * | 2005-11-25 | 2011-05-11 | 杨文龙 | Liquid capsule for treating diseases caused by pathogenic wind hot and its preparing method |
| CN101019949B (en) * | 2006-02-14 | 2011-09-14 | 清华大学 | Qingkailing preparation for great transfusion and its preparation process |
| CN102974124A (en) * | 2011-11-30 | 2013-03-20 | 洛阳惠中兽药有限公司 | Application of reduced pressure reflux technology in traditional Chinese medicine extraction |
| CN102974124B (en) * | 2011-11-30 | 2015-01-14 | 洛阳惠中兽药有限公司 | Application of reduced pressure reflux technology in traditional Chinese medicine extraction |
| CN114042105A (en) * | 2021-12-27 | 2022-02-15 | 广州白云山明兴制药有限公司 | Preparation process of traditional Chinese medicine compound oral liquid |
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