CN1686175A - Natural calculus bovis in vitro breeding method - Google Patents
Natural calculus bovis in vitro breeding method Download PDFInfo
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- CN1686175A CN1686175A CNA2005100461225A CN200510046122A CN1686175A CN 1686175 A CN1686175 A CN 1686175A CN A2005100461225 A CNA2005100461225 A CN A2005100461225A CN 200510046122 A CN200510046122 A CN 200510046122A CN 1686175 A CN1686175 A CN 1686175A
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Abstract
A process for externally culturing the natural ox gallstone includes collecting fresh ox gall, preparing the active components of gallstone, adding them collected ox gall, adding the active microbe composition, fermenting, preparing saturated solution of calcium oxide, preparing composite cholerythrin calcium, preparing ox gallstone, and examining quality.
Description
Affiliated field
The present invention relates to add the Calculus Bovis composition in Chinese medicine the pharmaceutical technology field, particularly Fel Bovis seu Bubali, use effective compound microorganism ferments, insert the natural calculus bovis in vitro breeding method of the container that slowly runs.
Background technology
Calculus Bovis is exsiccant Calculus Bovis, is famous and precious Chinese medicine material.Because natural gallstone formation rate is extremely low in the cattle body, so Calculus Bovis medicine source scarcity, dependence on import costs an arm and a leg.The present domestic 650 kinds of patent medicine that contain the Calculus Bovis composition that have add to 42 kinds of Chinese patent medicines again and ban use of the artificial Calculus Bovis, so the natural Calculus Bovis demand is big, existing output far can not satisfy the demand of China's national medicine industrial development.In order to change the situation of natural Calculus Bovis medicine source scarcity, many institution of higher learning, R﹠D institution and enterprise successively carry out big quantity research over domestic nearly 20 years, have all obtained some progress at different times, obtain many scientific achievements, have also declared nearly 10 patents simultaneously.Make a general survey of these achievements and patent, summarize to get up can be divided into five classes: the one, induce in cattle gallbladder with operation; The 2nd, in the cattle body, bury cultivating bottle and lure; The 3rd, with microorganism and bile direct fermentation; The 4th, endo-medicine is induced; Though some applies for a patent above-mentioned these technology, because of Calculus Bovis form low, can't industrialization development; Though what have has declared product batch number, can't accomplish scale production; The 5th kind of method is to utilize natural Fel Bovis seu Bubali in the medicinal Calculus Bovis method of animal cattle in vitro cultivation, just really realized industrialization production, realized the purpose of In vitro culture cholelithiasis.Yet; at present preparation natural Calculus Bovis fermentative microorganism multiselect is with escherichia coli or enterococcus and Bacillus proteus etc.; and need distinctive Calculus Bovis cultivating machine; and patent protection arranged; contain the pyrogen material in the goods more; and the product of making can not be used for quiet notes raw material, and technology is comparatively complicated, and product quality is still unstable.
Summary of the invention
The present invention seeks at the prior art deficiency, is to add the Calculus Bovis composition in the Fel Bovis seu Bubali, uses effective compound microorganism ferments, inserts the natural calculus bovis in vitro breeding method of the container that slowly runs.
Natural calculus bovis in vitro breeding method:
1, collects fresh Fel Bovis seu Bubali;
2, form Calculus Bovis active ingredient (seeing Table 1);
3, the Calculus Bovis active ingredient adds in the Fel Bovis seu Bubali, forms mixed liquor;
4, prepare effective complex microorganism preparations;
5, get weight portion mixed liquor 8000-12000, effective complex microorganism preparations bacterium liquid 400-600, the ferment tank of packing in proportion;
6, make saturated calcium oxide solution;
7, prepare compound calcium bilirubinate;
8, preparation In vitro Calculus Bovis (Calculus Bovis);
9, external cultivation natural Calculus Bovis quality arbitration;
10, in vitro cultivation natural Calculus Bovis packing and preservation.
This is that we are through 20 years of researches and tracking; consult the interrelated data that domestic and international a large amount of people and animal cholelithiasis form principle; on the basis of experimental in conjunction with oneself; according to the content characteristics that natural Calculus Bovis character, structure, composition are formed, the Calculus Bovis forming process is developed in the analogue body.It is relevant rudimentary subject knowledges such as microbiology, biochemistry, organic chemistry, inorganic chemistry, physics to be converged with being one, change bile and relevant active ingredient character, make it to become biliary calcules and bile, through overregulating PH, under suitable temperature through the running of low speed rotation container, form the aminoacid hydrogen bond, the albumen distortion is solidified, facilitate the core of calculus.The activity centre of core is because electrostatic attraction, make into calcium bilirubinate salt particle in the biliary calcules and bile and bilirubin high polymer granule to center deposition, optional general coating pan of this low speed rotation container or low speed large capacity centrifuge do not need special cultivating machine, under the situation of one-directional rotation, quicken big organic substance deposit seed is played bridging action, the function that is linear polymer is to particulate adsorption, be netted absorption layer by layer, increase gradually, thereby reach the purpose that forms Fel Bovis seu Bubali red pigment calcium calculus.
The technology of the present invention system uniqueness, technology is simple and clear, and the efficient height is stable and controllable for quality, can realize large-scale industrialized production; With regard to its drug effectiveness, safety, stable aspect, basic identical with the formation mechanism of the cholelithiasis of in the cattle body, cultivating, its character, structure, composition and even aminoacid, micronutrient levels and natural Calculus Bovis basically identical.Its characteristics are: the first, and can large-scale production with general coating pan or Bradytelic centrifugation of the large capacity machine etc.; The second, the Calculus Bovis active ingredient can oneself be extracted preparation, reduces cost; The 3rd, effectively complex microorganism preparations replaces microorganisms such as escherichia coli, enterococcus and enterobacteria, and these microorganisms all are useful ecological bacterium, not only strain is many, function entirely, action range, and sweat do not produce endotoxin, quality better, safety range is big.With the direct preparing natural Calculus Bovis intravenous fluid of this product, quality can be guaranteed; The 4th, raw material adopts lactic acid to handle, the product good water solubility, and inorganic composition is few.
Effectively complex microorganism preparations is a kind of novel complex microorganism preparations; be brown translucent liquid; pH value is between 3.5-4.5; form by the compound symbiosis culture of many normal effective microbes; do not contain any chemical harmful substance; have no side effect, free from environmental pollution, be to make Fel Bovis seu Bubali and Calculus Bovis active ingredient form the committed step of Calculus Bovis through biofermentation.The experiment proved that to be better than present general escherichia coli, Bacillus proteus and enterococcus etc., have easy, practical, effect is wide, fermentation finished thoroughly1, efficient is high and don't produce characteristics such as endotoxin.It is that the effective microbe of numerous species is used as a function colony that calculus bovis in vitro breeding uses effective complex microorganism preparations; it is compared with general microorganism formulation not only possesses multiple functional advantage, and has also shown high-tech content at itself.
Description of drawings
Accompanying drawing 1 of the present invention is a Calculus Bovis active ingredient composition table.
Embodiment
Example 1, natural calculus bovis in vitro breeding method
(1) collects fresh bovine bile
Collect the gallbladder that cattle is butchered the back fresh bovine in the cattle slaughterhouse, take out bile and pack into airtightly in the container, place under the 2-4 ℃ of condition and keep in Dark Place
(2) form Calculus Bovis active ingredient (seeing Table 1)
(3) the Calculus Bovis active ingredient adds in the Fel Bovis seu Bubali, forms mixed liquor
It is an amount of to get Fel Bovis seu Bubali, adds the Calculus Bovis active ingredient, detects taurine content, and regulating taurine content with bile is 6-10%, places 2-4 ℃ of preservation;
(4) prepare effective complex microorganism preparations
Effective composite microbial bacteria liquid 0.8-1.2, distilled water 48-52, glucose 2-3, sodium chloride 0.4-0.6, thiamine 0.8-1.2; Above-mentioned substance is fully mixed, add a cover in the container of packing into, not necessarily airtight, place 22-26 ℃ to make antibacterial breed activation, pH value 3.0-4.0 is advisable after waiting to ferment 10-14 days, places 2-8 ℃ of preservation standby, and the effective complex microorganism of this kind that ferments once more is called stock solution (mother solution) again as strain;
(5) get weight portion mixed liquor 8000-12000, effective complex microorganism preparations bacterium liquid 400-600, in this ratio, the ferment tank of packing into; Place fermentation in 22-26 ℃ of lasting 3-5 days, and stirred once, continue 25-35 minute at every turn, treat to be appropriateness when the fermented product pH value reaches 3-4, take out packing, place 2-8 ℃ of preservation standby every 4-8 hour.
(6) make saturated calcium oxide solution
It is some to get calcium oxide (quicklime), and adding distil water is an amount of constantly to be stirred, and makes it saturation, and static to collect supernatant standby.
(7) prepare compound calcium bilirubinate
Get the weight portion Fel Bovis seu Bubali 8000-12000 that ferments, add the saturated calcium oxide solution 20000-24000 of clarification, put in the retort,, also can replace stirring, continue 30-60 minute with 90-100 ℃ of steam heated, this method homogeneous heating; The reddish brown precipitation thing is taken out in the cooling back, adding distil water 8000-12000, and 2-4 time repeatedly, fully stir, each static 30-50 minute, remove supernatant, add bilirubin at last again its content is reached more than 46%, standby.
(8) preparation In vitro Calculus Bovis (Calculus Bovis)
Get compound calcium bilirubinate 4000-6000; adding distil water 5000-10000; zinc sulfate 4-8; mix fully to stir and make suspension; regulating PH with rare lactic acid is 5.4-5.8; after pH mixed up; compound calcium bilirubinate liquid suspension is placed medicinal coating pan of standard or low speed large capacity centrifuge; place 36-38 ℃ of greenhouse; keep the fast rate of rotation speed in slow earlier back, after persistent period 40-80 minute, after leaving standstill again 4-6 hour; taking-up has been cultivated the formation Calculus Bovis and has been placed 50 ℃ of hothouse inner dryings, can obtain 1400-1600 in vitro cultivation ball piece shape natural Calculus Bovis.
(9) external cultivation natural Calculus Bovis quality arbitration
" quality will be higher than pharmacopeia for Chinese pharmacopoeia execution, internal control to press 2000
(10) in vitro cultivation natural Calculus Bovis packing and preservation.
The in vitro cultivation natural Calculus Bovis of making, assay approval reach that " after the Chinese pharmacopoeia version standard in 2000, pack, specification size, description are wanted standard up to specification.As commodity market circulation, must show the quality inspection report of national medicine inspection statutory machinery, large-scale production must have trade mark and approval produces code and meets GMP and produces external Calculus Bovis workshop, and will be through the authentication of national pharmaceutical control and administration department; Preservation condition is in shading, and is airtight, puts shady and cool dry place, protection against the tide, the anti-pressure.
Example 2, the preparation natural Calculus Bovis nutrient formulation of effective microbe: 1 kilogram of effective composite microbial bacteria liquid, 50 kilograms of distilled water, 2.5 kilograms of glucoses, 0.5 kilogram in sodium chloride, 1 kilogram of thiamine.
Example 3, effective microbe and Fel Bovis seu Bubali mixed culture fermentation: get weight portion mixed liquor 8000-12000 milliliter, effective complex microorganism preparations bacterium liquid 400-600 milliliter, the ferment tank of packing in proportion; Place fermentation in 22-26 ℃ of lasting 3-5 days, and stirred once, continue 25-35 minute at every turn, treat to be appropriateness when the fermented product pH value reaches 3-4, take out packing, place 2-8 ℃ of preservation standby every 4-8 hour.
Example 4, prepare compound calcium bilirubinate; Get the weight portion Fel Bovis seu Bubali 8000-12000 milliliter that ferments, add the saturated calcium oxide solution 20000-24000 milliliter of clarification, put in the retort,, also can replace stirring, continue 30-60 minute with 90-100 ℃ of steam heated, this method homogeneous heating; The reddish brown precipitation thing is taken out in the cooling back, adding distil water 8000-12000 milliliter, and 2-4 time repeatedly, fully stir, each static 30-50 minute, remove supernatant, add bilirubin at last again its content is reached more than 46%, standby.
Example 5, preparation In vitro Calculus Bovis (Calculus Bovis); Get compound calcium bilirubinate 4000-6000 milliliter; adding distil water 5000-10000 milliliter; zinc sulfate 4-8; mix fully to stir and make suspension; regulating PH with rare lactic acid is 5.4-5.8; after pH mixed up; compound calcium bilirubinate liquid suspension is placed medicinal coating pan of standard or low speed large capacity centrifuge; place 36-38 ℃ of greenhouse; keep the fast rate of rotation speed in slow earlier back, after persistent period 40-80 minute, after leaving standstill again 4-6 hour; taking-up has been cultivated the formation Calculus Bovis and has been placed 50 ℃ of hothouse inner dryings, can obtain 1400-1600 gram in vitro cultivation ball piece shape natural Calculus Bovis.
Example 6, get 10000 milliliters of the Calculus Bovis that contains taurine 8% and Fel Bovis seu Bubali mixed liquors, inoculate 500 milliliters of effective composite microbial bacteria liquid, the fermentation tank of packing into control temperature is 24 ℃, continues fermentation in 4 days, and stirs once every 6 hours, and each continuing stirred 30 minutes; Treat that the fermented product pH value reaches at 3.5 o'clock and is appropriateness, add 22000 milliliters of saturated calcium oxide solution of clarification again, put in the retort or stainless steel cask, be heated to 95 ℃, continue 45 minutes with the steam carburator; After the cooling, take out the reddish brown precipitation thing, 10000 milliliters of reuse distilled water, 3 times repeatedly, fully stir, each static 40 minutes, remove supernatant, and then add bilirubin and make its content reach 46%; 7500 milliliters of last adding distil waters, zinc sulfate 6 grams mix fully to stir and make suspension, regulating pH value with rare lactic acid is 5.6, after pH value is mixed up, compound calcium bilirubinate suspension is placed the medicinal small-sized coating pan of standard for about 10000 milliliters, 2000 milliliters of every pot of loading amounts are put 37 ℃ of greenhouses; The maintenance elder generation fast rate of rotation speed in slow back after 40 minutes persistent period, leaves standstill and cultivated 5 hours, and taking-up has been cultivated the formation Calculus Bovis and put into 50 ℃ of hothouse inner dryings, can obtain every pot of 0.3 kilogram of in vitro cultivation natural Calculus Bovis.
Example 7; get 2000 milliliters of Calculus Bovis active ingredient Fel Bovis seu Bubali mixed liquors that contain 8% taurine; inoculating 100 milliliters of effective composite microbial bacteria liquid fermentation flask control temperature of packing into is 25 ℃; continue fermentation in 3 days; and once every stirring in 5 hours; each lasting 35 minutes; treat that the fermented product pH value reaches at 4 o'clock; add 4000 milliliters of saturated aqua calcises of clarification again; put water-soluble pot heating; control 100 ℃ and continue 45 minutes; the reddish brown precipitation thing is taken out in the cooling back; 2000 milliliters of reuse distilled water; twice repeatedly; fully stir, each static 30 minutes, remove supernatant; add bilirubin then; its content is reached more than 45%, 1200 milliliters of last adding distil waters, zinc sulfate 1.2 grams; be mixed into the mixing suspension; regulating pH value with rare lactic acid is 5.5, compound calcium bilirubinate suspension is placed the Bradytelic centrifugation of the large capacity machine for about 2000 milliliters, each centrifugal bottled 500 milliliters; centrifugal in 36 ℃ of greenhouses; slow earlier back is fast, continues 45 minutes, static again 4 hours; take out the calculus of cultivating shaping and put into 50 ℃ of drying baker dryings, can obtain the spherical block natural Calculus Bovis of the in vitro cultivation of each centrifuge bottle 15 gram.
Claims (10)
1, natural calculus bovis in vitro breeding method:
(1) collects fresh Fel Bovis seu Bubali;
(2) form the Calculus Bovis active ingredient;
(3) the Calculus Bovis active ingredient adds in the Fel Bovis seu Bubali, forms mixed liquor;
(4) prepare effective complex microorganism preparations;
(5) get weight portion mixed liquor 8000-12000, effective complex microorganism preparations bacterium liquid 400-600, in this ratio, the ferment tank of packing into;
(6) make saturated calcium oxide solution;
(7) prepare compound calcium bilirubinate;
(8) preparation In vitro Calculus Bovis;
(9) external cultivation natural Calculus Bovis quality arbitration;
(10) in vitro cultivation natural Calculus Bovis packing and preservation.
2, according to the described natural calculus bovis in vitro breeding method of claim 1:
(1) collects fresh Fel Bovis seu Bubali;
Collect the gallbladder that cattle is butchered the back fresh bovine in the cattle slaughterhouse, take out bile and pack into airtightly in the container, place under the 2-4 ℃ of condition and keep in Dark Place;
(2) form the Calculus Bovis active ingredient;
(3) the Calculus Bovis active ingredient adds in the Fel Bovis seu Bubali, forms mixed liquor;
It is an amount of to get Fel Bovis seu Bubali, adds the Calculus Bovis active ingredient, detects taurine content, and regulating taurine content with bile is 6-10%, places 2-4 ℃ of preservation;
(4) prepare effective complex microorganism preparations;
Effective composite microbial bacteria liquid 0.8-1.2, distilled water 48-52, glucose 2-3, sodium chloride 0.4-0.6, thiamine 0.8-1.2; Above-mentioned substance is fully mixed, add a cover in the container of packing into, not necessarily airtight, place 22-26 ℃ to make antibacterial breed activation, pH value 3.0-4.0 is advisable after waiting to ferment 10-14 days, places 2-8 ℃ of preservation standby, and the effective complex microorganism of this kind that ferments once more is called stock solution again as strain;
(5) get weight portion mixed liquor 8000-12000, effective complex microorganism preparations bacterium liquid 400-600, in this ratio, the ferment tank of packing into; Place fermentation in 22-26 ℃ of lasting 3-5 days, and stirred once, continue 25-35 minute at every turn, treat to be appropriateness when the fermented product pH value reaches 3-4, take out packing, place 2-8 ℃ of preservation standby every 4-8 hour;
(6) make saturated calcium oxide solution;
It is some to get calcium oxide, and adding distil water is an amount of constantly to be stirred, and makes it saturation, and static to collect supernatant standby;
(7) prepare compound calcium bilirubinate;
Get the Fel Bovis seu Bubali 8000-12000 that ferments, add the saturated calcium oxide solution 20000-24000 of clarification, put in the retort,, also can replace stirring, continue 30-60 minute with 90-100 ℃ of steam heated, this method homogeneous heating; The reddish brown precipitation thing is taken out in the cooling back, adding distil water 8000-12000, and 2-4 time repeatedly, fully stir, each static 30-50 minute, remove supernatant, add bilirubin at last again its content is reached more than 46%, standby;
(8) preparation In vitro Calculus Bovis;
Get compound calcium bilirubinate 4000-6000, adding distil water 5000-10000, zinc sulfate 4-8, mix fully to stir and make suspension, regulating PH with rare lactic acid is 5.4-5.8, after PH mixed up, compound calcium bilirubinate liquid suspension is placed the medicinal coating pan of standard or low speed large capacity centrifuge or biological culture shaking table, place 36-38 ℃ of greenhouse, keep the fast rate of rotation speed in slow earlier back, after persistent period 40-80 minute, after leaving standstill again 4-6 hour, taking-up has been cultivated the formation Calculus Bovis and has been placed 50 ℃ of hothouse inner dryings, can obtain 1400-1600 in vitro cultivation ball piece shape natural Calculus Bovis;
(9) external cultivation natural Calculus Bovis quality arbitration;
" quality will be higher than pharmacopeia for Chinese pharmacopoeia execution, internal control to press 2000;
(10) in vitro cultivation natural Calculus Bovis packing and preservation;
The in vitro cultivation natural Calculus Bovis of making, assay approval reach that " after the Chinese pharmacopoeia version standard in 2000, pack, specification size, description are wanted standard up to specification.
3, according to the described natural calculus bovis in vitro breeding method of claim 1, content of bilirubin calculates by 100%, and material purity is not less than 60% 0.6-1.0; The 46-82 of bile pigments calcium salt material purity content of bilirubin more than 10%; Bile acid content calculates by 100%, and material purity is at the 11.5-14.5 more than 60%; The 11.5-14.5 of Hyodeoxycholic Acid material purity more than m.p140 ℃; The 1-3 of cholesterol material purity more than m.p150 ℃; The 0.8-1.2 of magnesium sulfate pharmaceutical grade; The 0.5-0.9 of ferrous sulfate pharmaceutical grade; The 1.5-3.5 of calcium hydrogen phosphate pharmaceutical grade, taurine 2-4.
4, according to claim 1 or 3 described natural calculus bovis in vitro breeding method, content of bilirubin calculates by 100%, material purity be not less than 60% 0.8; Bile pigments calcium salt material purity content of bilirubin 64 more than 10%; Bile acid content calculates by 100%, and material purity is at 13 more than 60%; Hyodeoxycholic Acid material purity 13 more than m.p140 ℃; Cholesterol material purity 2 more than m.p150 ℃; 1 of magnesium sulfate pharmaceutical grade; 0.7 of ferrous sulfate pharmaceutical grade; 2.5 of calcium hydrogen phosphate pharmaceutical grade; Taurine 3.
5, according to claim 1 natural calculus bovis in vitro breeding method, the Calculus Bovis active ingredient is formed: 1 kilogram of effective composite microbial bacteria liquid, 50 kilograms of distilled water, 2.5 kilograms of glucoses, 0.5 kilogram in sodium chloride, 1 kilogram of thiamine.
6,, get weight portion mixed liquor 8000-12000 milliliter, effective complex microorganism preparations bacterium liquid 400-600 milliliter, the ferment tank of packing in proportion according to claim 1 natural calculus bovis in vitro breeding method; Place fermentation in 22-26 ℃ of lasting 3-5 days, and stirred once, continue 25-35 minute at every turn, treat to be appropriateness when the fermented product pH value reaches 3-4, take out packing, place 2-8 ℃ of preservation standby every 4-8 hour.
7,, prepare compound calcium bilirubinate according to claim 1 natural calculus bovis in vitro breeding method; Get the weight portion Fel Bovis seu Bubali 8000-12000 milliliter that ferments, add the saturated calcium oxide solution 20000-24000 milliliter of clarification, put in the retort,, also can replace stirring, continue 30-60 minute with 90-100 ℃ of steam heated, this method homogeneous heating; The reddish brown precipitation thing is taken out in the cooling back, adding distil water 8000-12000 milliliter, and 2-4 time repeatedly, fully stir, each static 30-50 minute, remove supernatant, add bilirubin at last again its content is reached more than 46%, standby.
8, according to claim 1 natural calculus bovis in vitro breeding method, the preparation In vitro Calculus Bovis; Get compound calcium bilirubinate 4000-6000 milliliter; adding distil water 5000-10000 milliliter; zinc sulfate 4-8; mix fully to stir and make suspension; regulating PH with rare lactic acid is 5.4-5.8; after PH mixed up; compound calcium bilirubinate liquid suspension is placed medicinal coating pan of standard or low speed large capacity centrifuge; place 36-38 ℃ of greenhouse; keep the fast rate of rotation speed in slow earlier back, after persistent period 40-80 minute, after leaving standstill again 4-6 hour; taking-up has been cultivated the formation Calculus Bovis and has been placed 50 ℃ of hothouse inner dryings, can obtain 1400-1600 milliliter in vitro cultivation ball piece shape natural Calculus Bovis.
9, according to claim 1 natural calculus bovis in vitro breeding method, get 10000 milliliters of the Calculus Bovis that contains taurine 8% and Fel Bovis seu Bubali mixed liquors, inoculate 500 milliliters of effective composite microbial bacteria liquid, the fermentation tank of packing into control temperature is 24 ℃, continue fermentation in 4 days, and every stirring in 6 hours once, each continuing stirred 30 minutes; Treat that the fermented product pH value reaches at 3.5 o'clock and is appropriateness, add 22000 milliliters of saturated calcium oxide solution of clarification again, put in the retort or stainless steel cask, be heated to 95 ℃, continue 45 minutes with the steam carburator; After the cooling, take out the reddish brown precipitation thing, 10000 milliliters of reuse distilled water, 3 times repeatedly, fully stir, each static 40 minutes, remove supernatant, and then add bilirubin and make its content reach 46%; 7500 milliliters of last adding distil waters, zinc sulfate 6 grams mix fully to stir and make suspension, regulating pH value with rare lactic acid is 5.6, after pH value is mixed up, compound calcium bilirubinate suspension is placed the medicinal small-sized coating pan of standard for about 10000 milliliters, 2000 milliliters of every pot of loading amounts are put 37 ℃ of greenhouses; The maintenance elder generation fast rate of rotation speed in slow back after 40 minutes persistent period, leaves standstill and cultivated 5 hours, and taking-up has been cultivated the formation Calculus Bovis and put into 50 ℃ of hothouse inner dryings, can obtain every pot of 0.3 kilogram of in vitro cultivation natural Calculus Bovis.
10; according to claim 1 natural calculus bovis in vitro breeding method; get 2000 milliliters of Calculus Bovis active ingredient Fel Bovis seu Bubali mixed liquors that contain 8% taurine; inoculating 100 milliliters of effective composite microbial bacteria liquid fermentation flask control temperature of packing into is 25 ℃; continue fermentation in 3 days; and once every stirring in 5 hours; each lasting 35 minutes; treat that the fermented product pH value reaches at 4 o'clock; add 4000 milliliters of saturated aqua calcises of clarification again; put water-soluble pot heating; control 100 ℃ and continue 45 minutes; the reddish brown precipitation thing is taken out in the cooling back; 2000 milliliters of reuse distilled water, repeatedly twice, fully stir; each static 30 minutes; remove supernatant, add bilirubin then, its content is reached more than 45%; 1200 milliliters of last adding distil waters; zinc sulfate 1.2 grams are mixed into the mixing suspension, and regulating pH value with rare lactic acid is 5.5; compound calcium bilirubinate suspension is placed the Bradytelic centrifugation of the large capacity machine for about 2000 milliliters; each centrifugal bottled 500 milliliters, centrifugal in 36 ℃ of greenhouses, slow earlier back is fast; continue 45 minutes; static again 4 hours, take out the calculus of cultivating shaping and put into 50 ℃ of drying baker dryings, can obtain the spherical block natural Calculus Bovis of the in vitro cultivation of each centrifuge bottle 15 gram.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101284016B (en) * | 2008-06-04 | 2012-03-14 | 江苏华阳制药有限公司 | Bezoar vitro bred and solid oral preparation containing the bezoar |
| CN102429927A (en) * | 2011-12-21 | 2012-05-02 | 武汉大鹏药业有限公司 | Method for preparing cow gallstone |
| CN103690563A (en) * | 2013-12-08 | 2014-04-02 | 王昌勇 | Method for cultivating bezoar in vitro |
| CN111184589A (en) * | 2020-02-27 | 2020-05-22 | 四川农业大学 | Detachable bezoar bed and bezoar cultivation method |
| CN112641806A (en) * | 2021-02-04 | 2021-04-13 | 平措绕吉 | Artificial bezoar and preparation method thereof |
| CN114480020A (en) * | 2022-02-23 | 2022-05-13 | 上海云洛生物技术有限公司 | Method for biologically synthesizing ambergris |
| CN116392509A (en) * | 2023-06-08 | 2023-07-07 | 内蒙古为旭生物科技有限公司 | Method for inducing cow to form bezoar |
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2005
- 2005-03-29 CN CNA2005100461225A patent/CN1686175A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101284016B (en) * | 2008-06-04 | 2012-03-14 | 江苏华阳制药有限公司 | Bezoar vitro bred and solid oral preparation containing the bezoar |
| CN102429927A (en) * | 2011-12-21 | 2012-05-02 | 武汉大鹏药业有限公司 | Method for preparing cow gallstone |
| CN103690563A (en) * | 2013-12-08 | 2014-04-02 | 王昌勇 | Method for cultivating bezoar in vitro |
| CN103690563B (en) * | 2013-12-08 | 2017-01-18 | 王昌勇 | Method for cultivating bezoar in vitro |
| CN111184589A (en) * | 2020-02-27 | 2020-05-22 | 四川农业大学 | Detachable bezoar bed and bezoar cultivation method |
| CN112641806A (en) * | 2021-02-04 | 2021-04-13 | 平措绕吉 | Artificial bezoar and preparation method thereof |
| CN114480020A (en) * | 2022-02-23 | 2022-05-13 | 上海云洛生物技术有限公司 | Method for biologically synthesizing ambergris |
| CN116392509A (en) * | 2023-06-08 | 2023-07-07 | 内蒙古为旭生物科技有限公司 | Method for inducing cow to form bezoar |
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