CN1347700A - Method of in vitro culturing medicine ox gallstone with natural ox gall - Google Patents
Method of in vitro culturing medicine ox gallstone with natural ox gall Download PDFInfo
- Publication number
- CN1347700A CN1347700A CN01133550A CN01133550A CN1347700A CN 1347700 A CN1347700 A CN 1347700A CN 01133550 A CN01133550 A CN 01133550A CN 01133550 A CN01133550 A CN 01133550A CN 1347700 A CN1347700 A CN 1347700A
- Authority
- CN
- China
- Prior art keywords
- seu bubali
- bovis seu
- calculus
- fermentation
- fel bovis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 13
- 238000000338 in vitro Methods 0.000 title claims description 7
- 208000001130 gallstones Diseases 0.000 title abstract description 6
- 239000003814 drug Substances 0.000 title description 8
- 238000012258 culturing Methods 0.000 title 1
- BPYKTIZUTYGOLE-IFADSCNNSA-N Bilirubin Chemical compound N1C(=O)C(C)=C(C=C)\C1=C\C1=C(C)C(CCC(O)=O)=C(CC2=C(C(C)=C(\C=C/3C(=C(C=C)C(=O)N\3)C)N2)CCC(O)=O)N1 BPYKTIZUTYGOLE-IFADSCNNSA-N 0.000 claims abstract description 36
- 238000003756 stirring Methods 0.000 claims abstract description 17
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 claims abstract description 16
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 claims abstract description 12
- 229960002471 cholic acid Drugs 0.000 claims abstract description 12
- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000006228 supernatant Substances 0.000 claims abstract description 6
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 claims abstract description 5
- 229960003964 deoxycholic acid Drugs 0.000 claims abstract description 5
- 229960003080 taurine Drugs 0.000 claims abstract description 5
- 239000002131 composite material Substances 0.000 claims abstract description 4
- 241000894006 Bacteria Species 0.000 claims abstract description 3
- 159000000007 calcium salts Chemical class 0.000 claims abstract description 3
- 229910017053 inorganic salt Inorganic materials 0.000 claims abstract description 3
- 229920006395 saturated elastomer Polymers 0.000 claims abstract description 3
- 238000000855 fermentation Methods 0.000 claims description 26
- 230000004151 fermentation Effects 0.000 claims description 26
- -1 compound calcium bilirubinate Chemical class 0.000 claims description 15
- 241000283690 Bos taurus Species 0.000 claims description 13
- 210000000941 bile Anatomy 0.000 claims description 13
- 239000004575 stone Substances 0.000 claims description 12
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 claims description 11
- 239000004380 Cholic acid Substances 0.000 claims description 11
- 235000019416 cholic acid Nutrition 0.000 claims description 11
- 238000001556 precipitation Methods 0.000 claims description 10
- 239000000243 solution Substances 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
- 230000000927 lithogenic effect Effects 0.000 claims description 8
- 230000001580 bacterial effect Effects 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 241000588724 Escherichia coli Species 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 3
- 241000588769 Proteus <enterobacteria> Species 0.000 claims description 3
- 241000194033 Enterococcus Species 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 238000005352 clarification Methods 0.000 claims description 2
- 230000008030 elimination Effects 0.000 claims description 2
- 238000003379 elimination reaction Methods 0.000 claims description 2
- 238000002386 leaching Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 12
- 239000000203 mixture Substances 0.000 abstract description 8
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 abstract description 4
- 239000011575 calcium Substances 0.000 abstract description 4
- 229910052791 calcium Inorganic materials 0.000 abstract description 4
- 206010004542 Bezoar Diseases 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 230000001105 regulatory effect Effects 0.000 abstract 2
- 230000007721 medicinal effect Effects 0.000 abstract 1
- 239000002244 precipitate Substances 0.000 abstract 1
- 239000012266 salt solution Substances 0.000 abstract 1
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 6
- 230000007246 mechanism Effects 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 238000005138 cryopreservation Methods 0.000 description 3
- 210000000232 gallbladder Anatomy 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 3
- 235000019341 magnesium sulphate Nutrition 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000004659 sterilization and disinfection Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 3
- 229960001763 zinc sulfate Drugs 0.000 description 3
- 229910000368 zinc sulfate Inorganic materials 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- DGABKXLVXPYZII-UHFFFAOYSA-N Hyodeoxycholic acid Natural products C1C(O)C2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)CC2 DGABKXLVXPYZII-UHFFFAOYSA-N 0.000 description 2
- 239000004677 Nylon Substances 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- BRPQOXSCLDDYGP-UHFFFAOYSA-N calcium oxide Chemical class [O-2].[Ca+2] BRPQOXSCLDDYGP-UHFFFAOYSA-N 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- DGABKXLVXPYZII-SIBKNCMHSA-N hyodeoxycholic acid Chemical compound C([C@H]1[C@@H](O)C2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)CC1 DGABKXLVXPYZII-SIBKNCMHSA-N 0.000 description 2
- 238000011031 large-scale manufacturing process Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011785 micronutrient Substances 0.000 description 2
- 235000013369 micronutrients Nutrition 0.000 description 2
- 229920001778 nylon Polymers 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
- 206010010904 Convulsion Diseases 0.000 description 1
- 206010062717 Increased upper airway secretion Diseases 0.000 description 1
- 206010037660 Pyrexia Diseases 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000282894 Sus scrofa domesticus Species 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical class [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000015111 chews Nutrition 0.000 description 1
- 201000001883 cholelithiasis Diseases 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000036461 convulsion Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000000003 hoof Anatomy 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000001077 hypotensive effect Effects 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002350 laparotomy Methods 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 208000026435 phlegm Diseases 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000001054 red pigment Substances 0.000 description 1
- 239000003507 refrigerant Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 235000013599 spices Nutrition 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
Abstract
Natural ox gall is first taurine regulated and bacteria fermented to prepare fermented ox gall. The fermented ox gall is gall is then mixed with clear saturated calcium salt solution through stirring, heated to boil, filtered to obtain red brown precipitate, which is stirred, washed, filtered to eliminate supernatant and mixed with bilirubin, cholalic acid, deoxycholic acid, inorganic salt and other effective components to obtain composite bilirubin calcium. Fermented ox gall and bilirubin calcium in certain proportion are mixed and the pH of mixture is regulated to below 6.8 and directional off axis rotated to form gallstone, and the gallstone is rest cultured, taken out and dried. The said process can obtain ox gallstone product with the same characteristics, structure, component and medicinal effect as high quality natural bezoar.
Description
One, technical field
The invention belongs to the pharmaceutical technology field, particularly a kind of manufacture method of raw material of Chinese medicine medicine.
Two, background technology
Calculus Bovis is a Calculus Bovis, is precious Chinese crude drug.It has clear away heart-fire, eliminating phlegm, have one's ideas straightened out, function such as cool liver, endogenous wind stopping, heat-clearing and toxic substances removing.The many famous and precious Chinese patent medicines of China, as: cow-bezoar bolus for resurrection, LIUSHEN WAN etc. all is that principal agent is formulated with the Calculus Bovis.But natural gallstone formation rate only is 1～2% in the cattle body, and therefore, the medicine source is in great shortage, costs an arm and a leg, dependence on import, has not only spent a large amount of foreign exchanges, and has restricted China's traditional medicine industrial expansion.
Since 1956, throw oneself into artificial Calculus Bovis's research of the medical worker of China, the artificial Calculus Bovis was applied in the Chinese patent medicine prescription in 1971, and part has been alleviated the imbalance between supply and demand of natural Calculus Bovis.But this artificial Calculus Bovis mainly is bilirubin and a large amount of starch of cholic acid, Hyodeoxycholic Acid and denier grinds the powder that sieves and get into powder; it or not Calculus Bovis; its composition, structure, content and known various microcomponent are all too wide in the gap with natural Calculus Bovis; do not possess character, structure, composition, the content characteristics of natural Calculus Bovis, clinical efficacy is far away from natural Calculus Bovis.
Since the seventies in last century, the medical worker on ground such as Yunnan Province of China, Guangdong, Inner Mongol, the experimentation of Calculus Bovis cultivating has obtained success in the cattle on the hoof gallbladder.But need to carry out laparotomy ventrotomy and this operation more complicated to the cattle body, and 2-3 just can cut open the belly the taking-up nylon mesh, scrape pale brown color attachment.Gallstone formation rate is low, and output is little, and content of bilirubin is low, the quality instability.And this product is not a Calculus Bovis, but is deposited on pale brown lust pigment calcium granule and mucus on the nylon mesh.Scrape the formed powder in dry back.Therefore, can not large-scale production.
The beginning of the seventies in last century, the Cai Hongjiao doctor of Genneral Surgery of hospital of attached Tongji University of Wuhan Tongji Medical Univ. (being the present inventor) studies in reason and mechanism that human cholelithiasis is formed, and on the basis of succeeing, further carry out cultivating in the cattle gallbladder bile research of Calculus Bovis, obtained success and declared the patent of invention (application number 89100505.6) of " making the method for Calculus Bovis outside a kind of prosthesis " on January 25th, 1989 with this principle.The Calculus Bovis that utilizes this method to make, its outward appearance, structure, composition, content and natural Calculus Bovis are basic identical.But be limited to condition at that time, particularly version in 1985 " stipulate cholic acid, the content of bilirubin of Calculus Bovis, and therefore, the bilirubin of the Calculus Bovis that it is made, cholic acid equal size are lower by Chinese pharmacopoeia.Moreover, said method is except that the Fel Bovis seu Bubali of prescription, and how the preparation calcium bilirubinate replaces with Fel Sus domestica, and bile promptly can Cheng Shi without bacterial fermentation.But its Calculus Bovis of making contains Hyodeoxycholic Acid therefore.Nineteen ninety (" one one of Chinese pharmacopoeia nineteen ninety version) is regulation clearly: Calculus Bovis is a Calculus Bovis, contains bilirubin more than 35%, and cholic acid is more than 7%, and has stipulated corresponding discriminating and detection method.Therefore the Calculus Bovis made from said method can not satisfy the new " medicinal requirements that Chinese pharmacopoeia is stipulated Calculus Bovis.
Three, Fa Ming content
The present invention is on the basis of ZL89100505.6 patent of invention; according to new " Chinese pharmacopoeia require develop a kind of can large-scale production, can satisfy the new " method at the external making Calculus Bovis of animal cattle (being In vitro cultured Calculus Bovis) of Chinese pharmacopoeia requirement again.
Technical scheme of the present invention is as follows:
1. prepare the fermentation Fel Bovis seu Bubali:
(1) content of taurine in the natural Fel Bovis seu Bubali of adjusting makes it reach medicinal requirements.
(2) with escherichia coli, Bacillus proteus or enterococcal single bacterium liquid or its composite bacterial solution in right amount routinely inoculation method be inoculated in the natural Fel Bovis seu Bubali, natural Fel Bovis seu Bubali is carried out bacterial fermentation handles.
2. utilize the fermentation Fel Bovis seu Bubali to prepare compound calcium bilirubinate:
(1) takes out quantitative fermentation Fel Bovis seu Bubali, add the saturated calcium salt soln of clarification that 1-4 doubly measures, it is stirred, be heated to and boil, leaching reddish brown precipitation thing.
(2) in the reddish brown precipitation thing, add an amount of water it is stirred, washs, leave standstill back elimination supernatant, add an amount of bilirubin, cholic acid according to medicinal requirements again, behind deoxycholic acid and the inorganic salt, to its stir, mixing (can promptly with or drying for standby).
3. under 15 ℃ of-37 ℃ of conditions, utilize fermentation Fel Bovis seu Bubali and compound calcium bilirubinate at cattle in vitro cultivation Calculus Bovis:
(1) preparation lithogenic bile: the fermentation Fel Bovis seu Bubali that takes out quantitative volume, ratio in fermentation Fel Bovis seu Bubali amount 500～1500ml/kg adds compound calcium bilirubinate (compound calcium bilirubinate calculates by dried weight), add an amount of aqueous solution again, after fully stirring, leave standstill.
(2) cultivate Calculus Bovis:
1. regulate becoming the stone Fel Bovis seu Bubali to carry out PH, make its pH value below 6.8 with acid solution;
2. lithogenic bile is carried out the directed rotation of off-axis, treat that the spherical calculus of its class forms after, stop the rotation, leave standstill cultivation, take out calculus and carry out dried.
The mechanism that the present invention forms Calculus Bovis is: utilize the method for above-mentioned physics, biological and chemical, change the physics and chemistry of Fel Bovis seu Bubali, thereby formed one-tenth stone Fel Bovis seu Bubali.Lithogenic bile forms the albumen hydrogen bond through overregulating PH under the mild temperature condition, Partial Protein is out of shape, is solidified, and forms the core of calculus.The activity centre of core is because electrostatic attraction, makes calcium bilirubinate salt particle in the lithogenic bile and bilirubin high polymer granule to center deposition.Macromolecule organic confrontation deposit seed plays bridging action, and promptly the function of linear polymer is netted absorption layer by layer to particulate adsorption, increases gradually, thereby forms Fel Bovis seu Bubali red pigment calcium calculus.
According to the present invention at the Calculus Bovis of cattle in vitro cultivation; it is basic identical that its calculus mechanism and natural Calculus Bovis form mechanism; its character, structure, composition and even aminoacid; micronutrient levels is consistent with natural Calculus Bovis, its calmness, convulsion, bring down a fever, hypotensive effect is consistent with natural Calculus Bovis with intensity antibiotic, antiinflammation.Process conditions of the present invention are complete, and the active ingredient of its product and stable content thereof meet the existing " regulation of Chinese pharmacopoeia fully.Become stone efficient height, can carry out large-scale industrial production.Its production cost is significantly less than natural Calculus Bovis, has good social benefit and economic benefit.
Four, the specific embodiment
1. embodiment one:
(1) obtaining of natural Fel Bovis seu Bubali: take out fresh bovine bile, by the requirement of high-quality Calculus Bovis, regulate content of taurine and make it reach 7%, insert in the disinfection tank, cryopreservation is standby.
(2) preparation fermentation Fel Bovis seu Bubali: Fel Bovis seu Bubali is placed in the fermentation tank, inoculates the ratio of 100ml thalline solution, the composite bacterial solution of escherichia coli and Bacillus proteus is seeded in the Fel Bovis seu Bubali by every 10000ml Fel Bovis seu Bubali, ferment 80 hours standby.
(3) prepare compound calcium bilirubinate:
1. get fermentation Fel Bovis seu Bubali 20000ml adding 22000ml and clarify saturated calcium oxide solution, stir, boil, after the cooling, take out the reddish brown precipitation thing.
2. the reddish brown precipitation thing adds pure water 10000ml, fully stirs, and leaves standstill hypsokinesis and removes supernatant, adds bilirubin to its content and reaches more than 45%, adds zinc sulfate 3g, magnesium sulfate 3g again, and stirring, mixing are standby.
(4) under 37 ℃ of room temperature conditions, cultivate medicinal Calculus Bovis:
1. be prepared into the stone Fel Bovis seu Bubali: get fermentation Fel Bovis seu Bubali 1000ml and join in the compound calcium bilirubinate of 2kg for preparing through above-mentioned steps (3), add water 3000ml again, stir into suspension, standby.
2. produce Calculus Bovis: a. and with dilute sulfuric acid lithogenic bile is carried out the PH adjusting, making its pH value is 5.8; B. to becoming the stone Fel Bovis seu Bubali to carry out the directed rotation of off-axis, treat that the spherical calculus of class forms after, stop the rotation, leave standstill cultivation, take out and carry out dried, promptly get the Calculus Bovis finished product of about 800g.
Press dry product and calculate, it is 13% that above-mentioned Calculus Bovis finished product contains cholic acid, and containing bilirubin is 36%, and other composition and content thereof and natural Calculus Bovis basically identical meet the existing " regulation of Chinese pharmacopoeia.
2. embodiment two:
(1) obtaining of natural Fel Bovis seu Bubali: when killing cattle, under the condition of sterile working, win gallbladder; select free from admixture, not comtaminated, golden yellow person for use, inclining bile in disinfection tank, by the requirement of high-quality Calculus Bovis; regulate taurine, make its content to 6%, cryopreservation is standby.
(2) preparation fermentation Fel Bovis seu Bubali: Fel Bovis seu Bubali is placed in the fermentation tank, and the ratio in every 10000ml Fel Bovis seu Bubali inoculation 200ml thalline solution is seeded to escherichia coli in the Fel Bovis seu Bubali, ferments 96 hours, and is standby.
(3) prepare compound calcium bilirubinate:
1. get fermentation Fel Bovis seu Bubali 20000ml adding 60000ml and clarify saturated lime water, stir, boil, be cooled to 50 ℃, take out the reddish brown precipitation thing.
2. the reddish brown precipitation thing adds cold boiled water 10000ml, stirs, leaves standstill 30 minutes, and the supernatant that inclines adds bilirubin to its content and reaches more than 45%, adds cholic acid 120g again, deoxycholic acid 50g, magnesium sulfate 5g, and zinc sulfate 5g, stirring, mixing cool off standby.
(4) under 26 ℃ of room temperature conditions, cultivate medicinal Calculus Bovis:
1. be prepared into the stone Fel Bovis seu Bubali: get fermentation Fel Bovis seu Bubali 1000ml, join in the compound calcium bilirubinate of 1kg through above-mentioned steps (3) preparation, add the aqueous solution 3000ml that leaches in the compound calcium bilirubinate preparation process again, stir into suspension, make into the stone Fel Bovis seu Bubali.
2. produce Calculus Bovis: a. and regulate becoming the stone Fel Bovis seu Bubali to carry out PH with dilute hydrochloric acid, making its pH value is 6.B. utilize eccentric cranking machine (ZL99238345.5) to becoming the directed rotation of stone Fel Bovis seu Bubali, under the condition of 60 rev/mins of the speeds of rotation, rotated 20 minutes, promptly form the class round, leave standstill and cultivate after 4 hours, take out and carry out dried, promptly can be made into the Calculus Bovis finished product of about 800g.
Above-mentioned finished product is the class spherical form, and diameter 0.5～3cm, surperficial yellowish red color are to brownish red, and body is light, matter is crisp, and concentric laminated striation is arranged, and gas perfume (or spice), bitter in the mouth are then sweet, and refrigerant sense is arranged, and that chews is frangible, does not stick to one's teeth.Press dry product and calculate its cholic acid content 17%, content of bilirubin 36%, all the other compositions and even aminoacid, micronutrient levels and high-quality natural Calculus Bovis basically identical.Meet the existing " regulation of Chinese pharmacopoeia.
3. embodiment three:
(1) obtaining of natural Fel Bovis seu Bubali: take out fresh bovine bile, insert in the disinfection tank, cryopreservation is standby.
(2) preparation fermentation Fel Bovis seu Bubali: Fel Bovis seu Bubali is placed in the fermentation tank, and the ratio in 10000ml Fel Bovis seu Bubali inoculation 300ml thalline solution is seeded to enterococcus in the Fel Bovis seu Bubali, ferments 92 hours, and is standby.
(3) prepare compound calcium bilirubinate:
1. get fermentation Fel Bovis seu Bubali 29600ml, filter adding 88800ml and clarify saturated calcium oxide solution, stir, boil, the reddish brown precipitation thing is taken out in the cooling back.
2. the reddish brown precipitation thing adds water and fully stirs, and leaves standstill that hypsokinesis goes supernatant to add bilirubin to its content again to reach more than 45%, add cholic acid 100g, deoxycholic acid 40g, zinc sulfate 3g, magnesium sulfate 3g then, stir, mixing.
(4) under 15 ℃ of room temperature conditions, cultivate medicinal Calculus Bovis:
1. be prepared into the stone Fel Bovis seu Bubali: get fermentation Fel Bovis seu Bubali 1250ml and join in the compound calcium bilirubinate of 1kg for preparing through above-mentioned steps (3), add the aqueous solution 3000ml that leaches in the compound calcium bilirubinate preparation process again, stir into suspension, standby.
2. produce Calculus Bovis: (a) with dilute hydrochloric acid lithogenic bile is carried out PH and regulate, making its pH value is 5.6; (b) to becoming the stone Fel Bovis seu Bubali to carry out the directed rotation of off-axis, treat that the spherical calculus of class forms after, stop the rotation, leave standstill cultivation, take out and carry out dried, promptly make the Calculus Bovis finished product of about 1000g.
Press dry product and calculate, the cholic acid content of above-mentioned Calculus Bovis finished product is 15%, and content of bilirubin is 36.5%, and other composition and content thereof and natural Calculus Bovis basically identical meet the existing " regulation of Chinese pharmacopoeia.
Claims (1)
1. method of utilizing natural Fel Bovis seu Bubali at the medicinal Calculus Bovis of animal cattle in vitro cultivation is characterized in that:
(1) preparation fermentation Fel Bovis seu Bubali:
1. regulate the content of taurine in the natural Fel Bovis seu Bubali, make it reach medicinal requirements;
2. with single bacterium liquid or its composite bacterial solution of escherichia coli, enterococcus, Bacillus proteus
Be inoculated in the natural Fel Bovis seu Bubali, natural Fel Bovis seu Bubali carried out bacterial fermentation handle;
(2) utilize the fermentation Fel Bovis seu Bubali to prepare compound calcium bilirubinate:
1. take out quantitative fermentation Fel Bovis seu Bubali, add the saturated calcium salt soln of clarification of 1～4 times of amount, it is stirred, is heated to and boils, leaching reddish brown precipitation thing;
2. in the reddish brown precipitation thing, add an amount of water it is stirred, washs, leave standstill back elimination supernatant, again according to behind medicinal requirements adding an amount of bilirubin, cholic acid, deoxycholic acid and the inorganic salt, stirring, mixing;
(3) under 15 ℃～37 ℃ conditions, utilize fermentation Fel Bovis seu Bubali and compound calcium bilirubinate at cattle in vitro cultivation Calculus Bovis:
1. prepare lithogenic bile: take out the fermentation Fel Bovis seu Bubali of quantitative volume, add compound calcium bilirubinate, add an amount of aqueous solution again, leave standstill after fully stirring in the ratio of its 500～1500ml/kg.
2. cultivate Calculus Bovis: a. and be adjusted to the biliary PH of stone, make its pH value below 6.8 with acid solution; B. lithogenic bile is carried out the directed rotation of off-axis, treat that the spherical calculus of its class forms after, stop the rotation, leave standstill cultivation, take out calculus then and carry out dried.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011335505A CN1164276C (en) | 2001-10-12 | 2001-10-12 | Method of in vitro culturing medicine ox gallstone with natural ox gall |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011335505A CN1164276C (en) | 2001-10-12 | 2001-10-12 | Method of in vitro culturing medicine ox gallstone with natural ox gall |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1347700A true CN1347700A (en) | 2002-05-08 |
| CN1164276C CN1164276C (en) | 2004-09-01 |
Family
ID=4671911
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB011335505A Expired - Fee Related CN1164276C (en) | 2001-10-12 | 2001-10-12 | Method of in vitro culturing medicine ox gallstone with natural ox gall |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1164276C (en) |
Cited By (13)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101855027A (en) * | 2007-11-13 | 2010-10-06 | 麦克德米德股份有限公司 | Be used to control the composition and the method for galvanic corrosion in printed circuit boards |
| CN102302517A (en) * | 2011-08-25 | 2012-01-04 | 济南阿科普生物技术开发有限公司 | Method for producing calculus bovis with natural quality |
| 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 |
| CN102423320A (en) * | 2011-12-21 | 2012-04-25 | 武汉健民大鹏药业有限公司 | Method for preparing fermented oxgall by using oxgall |
| CN102429926A (en) * | 2011-12-21 | 2012-05-02 | 武汉健民大鹏药业有限公司 | Method for preparing composite calcium bilirubinate utilizing fermented oxgall |
| CN102429927A (en) * | 2011-12-21 | 2012-05-02 | 武汉大鹏药业有限公司 | Method for preparing ox gal stone |
| CN102512451A (en) * | 2011-12-21 | 2012-06-27 | 武汉健民大鹏药业有限公司 | Method for preparing cattle gall-stone by utilizing fermented oxgall and composite calcium bilirubinate |
| CN102512457A (en) * | 2011-12-21 | 2012-06-27 | 武汉健民大鹏药业有限公司 | Method for preparing composite calcium bilirubinate from ox gall |
| CN103316046A (en) * | 2013-06-09 | 2013-09-25 | 宁波优美肽生物科技有限公司 | Enzymatic production process for culturing calculus bovis in vitro |
| CN103690563A (en) * | 2013-12-08 | 2014-04-02 | 王昌勇 | Method for cultivating bezoar in vitro |
| CN114533763A (en) * | 2022-02-17 | 2022-05-27 | 肖德煊 | Novel method for culturing calculus bovis electric calculi in vitro |
| CN115851497A (en) * | 2022-09-16 | 2023-03-28 | 山东中医药大学 | Bovine bile resistant type bezoar transformation bacterium and application thereof |
| CN116059251A (en) * | 2023-03-07 | 2023-05-05 | 济南七贤堂生物科技有限公司 | Method for in vitro cultivation of natural quality bezoar by adopting biological enzyme |
-
2001
- 2001-10-12 CN CNB011335505A patent/CN1164276C/en not_active Expired - Fee Related
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101855027A (en) * | 2007-11-13 | 2010-10-06 | 麦克德米德股份有限公司 | Be used to control the composition and the method for galvanic corrosion in printed circuit boards |
| 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 |
| CN102302517A (en) * | 2011-08-25 | 2012-01-04 | 济南阿科普生物技术开发有限公司 | Method for producing calculus bovis with natural quality |
| CN102512457B (en) * | 2011-12-21 | 2013-06-19 | 武汉健民大鹏药业有限公司 | Method for preparing composite calcium bilirubinate from ox gall |
| CN102429927A (en) * | 2011-12-21 | 2012-05-02 | 武汉大鹏药业有限公司 | Method for preparing ox gal stone |
| CN102512451A (en) * | 2011-12-21 | 2012-06-27 | 武汉健民大鹏药业有限公司 | Method for preparing cattle gall-stone by utilizing fermented oxgall and composite calcium bilirubinate |
| CN102512457A (en) * | 2011-12-21 | 2012-06-27 | 武汉健民大鹏药业有限公司 | Method for preparing composite calcium bilirubinate from ox gall |
| CN102423320A (en) * | 2011-12-21 | 2012-04-25 | 武汉健民大鹏药业有限公司 | Method for preparing fermented oxgall by using oxgall |
| CN102429926A (en) * | 2011-12-21 | 2012-05-02 | 武汉健民大鹏药业有限公司 | Method for preparing composite calcium bilirubinate utilizing fermented oxgall |
| CN103316046B (en) * | 2013-06-09 | 2014-12-17 | 宁波优美肽生物科技有限公司 | Enzymatic production process for culturing calculus bovis in vitro |
| CN103316046A (en) * | 2013-06-09 | 2013-09-25 | 宁波优美肽生物科技有限公司 | Enzymatic production process for culturing calculus bovis in vitro |
| 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 |
| CN114533763A (en) * | 2022-02-17 | 2022-05-27 | 肖德煊 | Novel method for culturing calculus bovis electric calculi in vitro |
| CN115851497A (en) * | 2022-09-16 | 2023-03-28 | 山东中医药大学 | Bovine bile resistant type bezoar transformation bacterium and application thereof |
| CN115851497B (en) * | 2022-09-16 | 2024-03-19 | 山东中医药大学 | Ox bile-resistant bezoar transformation strain and application thereof |
| CN116059251A (en) * | 2023-03-07 | 2023-05-05 | 济南七贤堂生物科技有限公司 | Method for in vitro cultivation of natural quality bezoar by adopting biological enzyme |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1164276C (en) | 2004-09-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1164276C (en) | Method of in vitro culturing medicine ox gallstone with natural ox gall | |
| CN100361583C (en) | Composite nano anti-biotic material and preparation method | |
| CN101760497B (en) | Microorganisms or/and biological enzyme extracting method for plant anthocyanin | |
| CN108503044A (en) | A kind of cultivation factory water body flocculation oxygenation sterilization water purification agent and preparation method thereof | |
| CN102429927B (en) | Method for preparing ox gal stone | |
| CN106377433B (en) | A kind of antibiotic property tooth root canal filling material and preparation method thereof | |
| CN103301151A (en) | Silver-iodide-doped bioactive glass as well as preparation method and application of silver-iodide-doped bioactive glass | |
| CN103262980A (en) | Selenium germanium chromium elements-enriched vitamin spirulina health-care rice flour | |
| CN113044995A (en) | Plant fermentation extract for freshwater aquaculture and preparation method thereof | |
| CN101711766B (en) | Medicine for treating white spots and rotten skin of turtle | |
| CN100340245C (en) | Bone strengthening oral liquor and its preparing method | |
| CN110272032A (en) | A kind of preparation method of magnesium doped hydroxyapatite drug bearing microsphere | |
| CN104825489A (en) | Silver-zinc powder-containing external gel and preparation method thereof | |
| CN115998962A (en) | Synthesis method of tissue-adhesive composite hydrogel and application of green sequence thereof in treatment of peri-implant inflammation | |
| CN113800650B (en) | Environment-friendly preparation for blue-green algae treatment and preparation method thereof | |
| CN107473359A (en) | A kind of functional mineral substance water and preparation method thereof | |
| CN102512451A (en) | Method for preparing cattle gall-stone by utilizing fermented oxgall and composite calcium bilirubinate | |
| CN107007633A (en) | A kind of probiotics polysaccharide composite formulation preparation method | |
| CN108464416A (en) | Natural toxin-expelling anti-oxidation health beverage | |
| CN113769162B (en) | Preparation method of polyvinylpyrrolidone iodine loaded biomimetic mineralized microspheres for treating infectious bone defects | |
| CN106893432A (en) | Zinc-rich selenium oyster shell whiting disease-prevention health nano inner wall coating | |
| CN110170007B (en) | Composition for inhibiting pigmentation in skin healing process and preparation method and application thereof | |
| RU2159630C1 (en) | Reparation and regeneration process stimulation agent | |
| CN106821861A (en) | Ferment zymoprotein toxin-expelling and face nourishing content in freckle cream | |
| CN1203878C (en) | Organic state micro element supplement agent |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: WUHAN JIANMIN DAPENG PHARMACEUTICAL CO., LTD. Free format text: FORMER OWNER: WUHAN DAPENG PHARMACEUTICAL INDUSTRY CO., LTD. Effective date: 20060303 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20060303 Address after: 430040 No. 15 Wu Nan Road, Dongxihu District, Hubei, Wuhan Patentee after: Wuhan key people Dapeng Pharmaceutical Co., Ltd. Address before: 430014 No. 102 Yanjiang Avenue, Han Han, Wuhan, Hubei Patentee before: Wuhan Dapeng Pharmaceutical Industry Co., Ltd. |
|
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20040901 Termination date: 20201012 |