CN1082409A - The extracting method of medical natural hirudiu - Google Patents
The extracting method of medical natural hirudiu Download PDFInfo
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- CN1082409A CN1082409A CN 92107076 CN92107076A CN1082409A CN 1082409 A CN1082409 A CN 1082409A CN 92107076 CN92107076 CN 92107076 CN 92107076 A CN92107076 A CN 92107076A CN 1082409 A CN1082409 A CN 1082409A
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- Prior art keywords
- hirudo
- hours
- hirudin
- solution
- slurry
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- 238000000034 method Methods 0.000 title claims abstract description 12
- 241000237903 Hirudo Species 0.000 claims abstract description 18
- 102000007625 Hirudins Human genes 0.000 claims abstract description 17
- 108010007267 Hirudins Proteins 0.000 claims abstract description 17
- WQPDUTSPKFMPDP-OUMQNGNKSA-N hirudin Chemical compound C([C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC=1C=CC(OS(O)(=O)=O)=CC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CCCCN)NC(=O)[C@H]1N(CCC1)C(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H]1NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(O)=O)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H]2CSSC[C@@H](C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(=O)N[C@H](C(NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N2)=O)CSSC1)C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=2C=CC(O)=CC=2)NC(=O)[C@@H](NC(=O)[C@@H](N)C(C)C)C(C)C)[C@@H](C)O)CSSC1)C(C)C)[C@@H](C)O)[C@@H](C)O)C1=CC=CC=C1 WQPDUTSPKFMPDP-OUMQNGNKSA-N 0.000 claims abstract description 17
- 229940006607 hirudin Drugs 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000002002 slurry Substances 0.000 claims abstract description 8
- 239000010413 mother solution Substances 0.000 claims abstract description 6
- 238000001556 precipitation Methods 0.000 claims abstract description 6
- 238000000502 dialysis Methods 0.000 claims abstract description 4
- 238000004090 dissolution Methods 0.000 claims abstract description 4
- 238000012869 ethanol precipitation Methods 0.000 claims abstract description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 14
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 6
- 239000000385 dialysis solution Substances 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 4
- 239000000243 solution Substances 0.000 claims description 4
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 238000004821 distillation Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000012362 glacial acetic acid Substances 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 2
- 238000004108 freeze drying Methods 0.000 claims description 2
- 238000010438 heat treatment Methods 0.000 claims description 2
- 239000012535 impurity Substances 0.000 claims description 2
- 239000011265 semifinished product Substances 0.000 claims 2
- 239000000284 extract Substances 0.000 abstract description 5
- 238000001291 vacuum drying Methods 0.000 abstract description 4
- 230000018044 dehydration Effects 0.000 abstract description 3
- 238000006297 dehydration reaction Methods 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 238000000605 extraction Methods 0.000 abstract description 2
- 239000000047 product Substances 0.000 abstract description 2
- 239000007795 chemical reaction product Substances 0.000 abstract 1
- 239000004615 ingredient Substances 0.000 abstract 1
- 238000005406 washing Methods 0.000 abstract 1
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000013543 active substance Substances 0.000 description 2
- 239000012043 crude product Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000012856 packing Methods 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N acetone Substances CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000011149 active material Substances 0.000 description 1
- 230000002429 anti-coagulating effect Effects 0.000 description 1
- 239000003146 anticoagulant agent Substances 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 238000001641 gel filtration chromatography Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 239000002893 slag Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The extracting method of medical natural hirudiu relates to the extraction of animal effective ingredient.The present invention adopts and extract hirudin from whole Hirudo, regulate pH value to 4.4-4.6 after earlier the Hirudo slurry being heated, the re-adjustment pH value to 6-7 of lowering the temperature again, centrifuge dehydration then, residue normal saline cyclic washing, mother solution dialysis back is used water dissolution with 95% above ethanol precipitation behind the precipitation vacuum drying, dialysis once more, last vacuum drying obtains elaboration.The hirudin process of extracting with the present invention is easy, and end product quality is qualified, and yield brings up to 4/1000ths, thereby greatly reduces product cost.
Description
The extracting method of medical natural hirudiu relates to the extraction of animal active ingredient.
Nearly one over 100 years, and people have affirmed that from being stung by Hirudo to inhale the phenomenon that The blood streamed down of back the Hirudo body contains anticoagulative substance.John B, professor Haycrafft of Wales country medical college find, the doctor trematodiasis is pharyngeal to have a heat-proof water solublity anticoagulant active material, F.Frany had separated purer similar active substance in 1903, and C, Jacoby were with its called after hirudin (hirudin) in 1904.Long-term next, owing to extract the resource-Hirudo of hirudin, though it is wide to distribute, rareness is concentrated, is difficult to again catch, and therefore can't put into production; Hirudin is again at the active substance that many special roles are medically arranged, state's exterior-applied liquid medicine-acetone mixed solution extracts, ethanol precipitation, cation exchange column and gel filtration chromatography method of purification extract refining hirudin, but this method complex steps, reagent cost an arm and a leg; Also have some major companies to drop into a huge sum of money, with the analog of gene engineering method recombinant natural hirudin, therefore the hirudin price used of every gram reagent up to 80,000 dollars about, and the home and abroad does not supply pharmaceutical hirudin at present.
Not only method is easy for the extracting method of the present invention's development, and reagent is cheap, and has improved the yield of hirudin.
The technical scheme that reaches the foregoing invention purpose is: integral body was smashed pulp to pieces after Hirudo was cleaned impurity such as removing earth, the Hirudo slurry is heated to 60-80 ℃, and kept 2-3 minute, the glacial acetic acid of adding 10% is adjusted pH value at 4.4-4.6 in the Hirudo slurry after heating, be cooled to 25-30 ℃ immediately, add 10% Na again
2CO
3Adjust pH value at 6-7, Hirudo is starched centrifugal collection mother solution, the normal saline lixiviate of residue adding 0.9% 2-3 time, collect whole lixiviating solution, with this mother solution dialysis 36 hours, changed water once every 12 hours, dialysis solution is with the ethanol precipitation more than 95%, adding 95% above ethanol earlier, to make its content be 61%, placed 12 hours, the supernatant that inclines, it is 85% that the ethanol that adds same concentration again makes its content, placed 24 hours, obtain hirudin precipitation, the precipitation cold drying gets crude product, and crude product was dialysed 16 hours after with an amount of second distillation water dissolution again, the sterilization of dialysis solution sucking filtration, elaboration is made in lyophilization.
The general hirudin that extracts is to get the head half to smash the back to pieces and extract, and the whole Hirudo that the present invention adopts, can make full use of like this Hirudo in the plain capsule of 11 couple of the intravital storage food of Hirudo divide must hirudin, thereby can improve the yield of hirudin greatly.
With 1 kilogram of Hirudo, all to smash to pieces and be muddy, the Hirudo slurry is heated to 75 ℃, keeps 3 minutes, with 10% glacial acetic acid accent pH value to 4.5, on the rocksly then is cooled to 30 ℃, uses 10%Na
2CO
3Transfer pH value to 6.5, the centrifuge dehydration of Hirudo slurry, collect mother solution, residue is washed with a small amount of normal saline, stir centrifuge dehydration, repeat washery slag once, collect three times solution, the bag filter of packing into was dialysed 36 hours, changed water once in per 12 hours, dialysis solution adds 95% above ethanol makes concentration reach 61%, precipitates 12 hours, collects the supernatant, the ethanol that adds same concentration again makes concentration reach 85%, precipitate 48 hours, collecting precipitation, vacuum drying under 30 ℃ of temperature conditions, remove the ethanol that contains, the exsiccant precipitation bag filter of packing into after with 100ml second distillation water dissolution was dialysed 16 hours, changed water once in 8 hours, dialysis solution gets 4.1 gram finished products with G6 funnel sucking filtration sterilization, filtrate vacuum drying.
The medical natural hirudiu process of extracting with the inventive method is simple, and the yield of hirudin brings up to 4/1000ths, thereby greatly reduces day production cost of trematodiasis element.
Claims (1)
1, a kind of extracting method of medical natural hirudiu is used the normal saline lixiviate, the invention is characterized in:
(1) with behind the impurity such as the clean removal of Hirudo earth, integral body is smashed into muddy to pieces,
(2) the Hirudo slurry is heated to 60-80 ℃, and keeps 2-3 minute,
(3) glacial acetic acid of adding 10% is adjusted pH value at 4.4-4.6 in the slurry of the Hirudo after heating, and is cooled to 25-30 ℃ immediately, adds 10%Na again
2CO
3Adjust pH value at 6-7,
(4) the Hirudo slurry is through centrifugal collection mother solution, and whole lixiviating solution are collected in the normal saline lixiviate successively of residue adding 0.9% 2-3 time,
(5) the mother solution dialysis is 36 hours, changes water once every 12 hours,
(6) dialysis solution is with 95% above ethanol precipitation, and adding 95% ethanol earlier, to make its content be 61%, placed 12 hours, and the supernatant that inclines, it is 85% that the ethanol that adds same concentration again makes its content, places 24 hours, obtains the hirudin precipitation,
(7) will precipitate cold drying and get semifinished product,
(8) semifinished product was dialysed 16 hours with an amount of second distillation water dissolution again, the sterilization of dialysis solution sucking filtration, and the solution lyophilization gets elaboration.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92107076 CN1082409A (en) | 1992-07-28 | 1992-07-28 | The extracting method of medical natural hirudiu |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 92107076 CN1082409A (en) | 1992-07-28 | 1992-07-28 | The extracting method of medical natural hirudiu |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1082409A true CN1082409A (en) | 1994-02-23 |
Family
ID=4942641
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 92107076 Pending CN1082409A (en) | 1992-07-28 | 1992-07-28 | The extracting method of medical natural hirudiu |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1082409A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102603890A (en) * | 2012-03-23 | 2012-07-25 | 韶关市颐林泉生物科技有限公司 | Method for extracting hirudin |
| CN102952189A (en) * | 2012-10-19 | 2013-03-06 | 旌德县振亚水产养殖有限公司 | Hirudin extraction bottle and hirudin extraction method |
| CN103417580A (en) * | 2013-09-04 | 2013-12-04 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN103784477A (en) * | 2014-02-08 | 2014-05-14 | 杭州桐君堂生物科技有限公司 | Preparation process of high-activity Hirudo micropowder for direct oral administration |
| CN105076586A (en) * | 2014-05-15 | 2015-11-25 | 钱如力 | A Polypeptide lucid ganoderma tea and preparation method thereof |
| CN106387309A (en) * | 2016-08-31 | 2017-02-15 | 西安若翰生物饲料有限公司 | High-efficient bio-fermented feed |
| CN107337729A (en) * | 2017-07-25 | 2017-11-10 | 重庆多普泰制药股份有限公司 | A kind of thick extracting method of hirudin |
| CN113073093A (en) * | 2021-04-25 | 2021-07-06 | 山东中医药大学 | Leech fibrinolytic active protein and preparation method thereof |
| CN116725928A (en) * | 2023-08-14 | 2023-09-12 | 广东尚品汇生物科技有限公司 | Anti-allergy acne-removing traditional Chinese medicine composition, and preparation method and application thereof |
-
1992
- 1992-07-28 CN CN 92107076 patent/CN1082409A/en active Pending
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102603890A (en) * | 2012-03-23 | 2012-07-25 | 韶关市颐林泉生物科技有限公司 | Method for extracting hirudin |
| CN102952189A (en) * | 2012-10-19 | 2013-03-06 | 旌德县振亚水产养殖有限公司 | Hirudin extraction bottle and hirudin extraction method |
| CN103417580A (en) * | 2013-09-04 | 2013-12-04 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN103417580B (en) * | 2013-09-04 | 2015-07-22 | 南宁市净雪皇生物工程有限公司 | Method for preparing poecilobdella manillensis medium-high activity anticoagulant crude product |
| CN103784477A (en) * | 2014-02-08 | 2014-05-14 | 杭州桐君堂生物科技有限公司 | Preparation process of high-activity Hirudo micropowder for direct oral administration |
| CN103784477B (en) * | 2014-02-08 | 2016-08-31 | 杭州桐君堂生物科技有限公司 | High activity is directly administered orally the manufacturing process of Hirudo micropowder |
| CN105076586A (en) * | 2014-05-15 | 2015-11-25 | 钱如力 | A Polypeptide lucid ganoderma tea and preparation method thereof |
| CN106387309A (en) * | 2016-08-31 | 2017-02-15 | 西安若翰生物饲料有限公司 | High-efficient bio-fermented feed |
| CN107337729A (en) * | 2017-07-25 | 2017-11-10 | 重庆多普泰制药股份有限公司 | A kind of thick extracting method of hirudin |
| CN113073093A (en) * | 2021-04-25 | 2021-07-06 | 山东中医药大学 | Leech fibrinolytic active protein and preparation method thereof |
| CN116725928A (en) * | 2023-08-14 | 2023-09-12 | 广东尚品汇生物科技有限公司 | Anti-allergy acne-removing traditional Chinese medicine composition, and preparation method and application thereof |
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