CN101088514A - Bee venom refining process - Google Patents
Bee venom refining process Download PDFInfo
- Publication number
- CN101088514A CN101088514A CNA2006100469560A CN200610046956A CN101088514A CN 101088514 A CN101088514 A CN 101088514A CN A2006100469560 A CNA2006100469560 A CN A2006100469560A CN 200610046956 A CN200610046956 A CN 200610046956A CN 101088514 A CN101088514 A CN 101088514A
- Authority
- CN
- China
- Prior art keywords
- precipitate
- venenum apis
- bee venom
- alcohol
- apis
- 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.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims abstract description 13
- 238000007670 refining Methods 0.000 title claims abstract description 11
- 239000003659 bee venom Substances 0.000 title abstract description 7
- 239000002244 precipitate Substances 0.000 claims abstract description 15
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 13
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000012535 impurity Substances 0.000 claims abstract description 6
- 239000000706 filtrate Substances 0.000 claims abstract description 4
- 238000004108 freeze drying Methods 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000007791 liquid phase Substances 0.000 claims abstract description 4
- 239000000725 suspension Substances 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims abstract description 3
- 241000241413 Propolis Species 0.000 claims abstract description 3
- 239000000908 ammonium hydroxide Substances 0.000 claims abstract description 3
- 229940069949 propolis Drugs 0.000 claims abstract description 3
- 238000000746 purification Methods 0.000 claims description 5
- 239000000284 extract Substances 0.000 claims description 4
- 239000012141 concentrate Substances 0.000 claims description 3
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 3
- 230000018044 dehydration Effects 0.000 claims description 3
- 238000006297 dehydration reaction Methods 0.000 claims description 3
- 229960004756 ethanol Drugs 0.000 claims description 3
- 238000012869 ethanol precipitation Methods 0.000 claims description 3
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 claims description 3
- 230000001476 alcoholic effect Effects 0.000 claims description 2
- 238000004821 distillation Methods 0.000 claims description 2
- 239000012153 distilled water Substances 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 238000000638 solvent extraction Methods 0.000 claims description 2
- 108010036176 Melitten Proteins 0.000 abstract description 5
- VDXZNPDIRNWWCW-JFTDCZMZSA-N melittin Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(N)=O)CC1=CNC2=CC=CC=C12 VDXZNPDIRNWWCW-JFTDCZMZSA-N 0.000 abstract description 5
- 238000001962 electrophoresis Methods 0.000 abstract description 4
- 238000001914 filtration Methods 0.000 abstract 1
- 235000012907 honey Nutrition 0.000 abstract 1
- 238000002386 leaching Methods 0.000 abstract 1
- 230000001376 precipitating effect Effects 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 239000002574 poison Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 241000040710 Chela Species 0.000 description 1
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000002435 venom Substances 0.000 description 1
- 231100000611 venom Toxicity 0.000 description 1
- 210000001048 venom Anatomy 0.000 description 1
Landscapes
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The bee venom refining process includes the following steps: dissolving coarse bee venom in alcohol to form suspension, filtering to eliminate propolis, honey and other impurity, centrifuging the filtrate and eliminating liquid phase, dewatering the precipitate with anhydrous alcohol, leaching the precipitate in water at 10+/-2 deg.c for 12-15 hr, precipitating the leached liquid with alcohol, eliminating water solution of alcohol, extracting the precipitate in ammonium hydroxide and n-butanol solution, distilling the extracted liquid, concentrating and freeze drying to obtain refined bee venom. The refined bee venom may be used in different clinical research and application fields, including preparing electrophoresis level melittin.
Description
Technical field
The present invention relates to the process for purification of Apis poison, particularly utilize refining Venenum apis to extract the refining of its Venenum apis of melittin.
Background technology
Apis poison (Bee Venom is called for short Venenum apis) is to be stored in the malicious capsule after being produced by bee venom, a kind of venom that when the Apis irriate, goes out from the chela faller gill, and Venenum apis is the compounding substances of a kind of pharmacological and biochemical activity high complexity, and extensive use is medically being arranged.But the clinical practice of Venenum apis is still very limited up till now, and reason mainly is that the composition of Venenum apis is numerous, refining purifying technique complexity, and cost is higher; Highly finished product still can not satisfy the clinical application requirements that waits special dimension, particularly can not satisfy the specification requirement of producing electrophoresis level melittin.
Summary of the invention
The objective of the invention is to overcome the deficiency that prior art exists, a kind of process for purification of Venenum apis is provided, the refining Venenum apis that this method obtains, the specification requirement that can satisfy clinical research and application and extract electrophoresis level melittin.
Process for purification provided by the invention is that thick Venenum apis is dissolved in impurity such as the propolis in the removal Venenum apis, Mel in the alcoholic solution, adopts the solvent extraction method again, obtains refining Venenum apis, and feature of the present invention is:
1) thick Venenum apis is added 95% ethanol and make suspension, filter and remove impurity, liquid phase is removed in the centrifugal back of filtrate, and precipitate adds the dehydrated alcohol dehydration, again the precipitate that proposes is added entry in 10 ± 2 ℃ of following lixiviate 12～15hr;
2) lixiviating solution ethanol precipitation is removed ethanol water, extracts in ammonium hydroxide that the precipitate adding is 0.15～0.2 mole and the butanol solution, and reject precipitate, distillation extraction liquid, concentrate carry out lyophilization and promptly obtain refining Venenum apis.
This method can be made with extra care the Venenum apis product by mass production, and craft science is reasonable, not only can satisfy the needs of more areas such as clinical research and application, can also satisfy the specification requirement of preparation electrophoresis level melittin.
Below by technology diagram and embodiment the present invention is described in detail.
Embodiment:
Get thick Venenum apis 100g, add 500ml95% ethanol and make suspension, jolting 4hr filters insoluble matter impurity such as removing the Apis sting; Remove liquid phase behind filtrate centrifugal (3000r/min) 20min, precipitate adds the dehydration of 300ml dehydrated alcohol, uses the 300mlPH=6.4 distilled water in 10 ± 2 ℃ of following lixiviate 12～15hr, reject precipitate the precipitate that proposes again, the lixiviating solution ethanol precipitation is removed ethanol water; Dissolve in precipitate adding 6000ml0.15 moles of hydrogen amine-oxides and the 4000ml butanol solution, jolting 2hr carries out liquid-solid separation, the reject precipitation, and extract distills the recovery n-butyl alcohol, and concentrate carries out lyophilization and promptly obtains refining Venenum apis.
Claims (2)
1, a kind of process for purification of Venenum apis is that thick Venenum apis is dissolved in impurity such as the propolis in the removal Venenum apis, Mel in the alcoholic solution, adopts the solvent extraction method again, obtains refining Venenum apis, and feature of the present invention is:
1) thick Venenum apis is added 95% ethanol and make suspension, filter and remove impurity, liquid phase is removed in the centrifugal back of filtrate, and precipitate adds the dehydrated alcohol dehydration, again the precipitate that proposes is added entry in 10 ± 2 ℃ of following lixiviate 12～15hr;
2) lixiviating solution ethanol precipitation is removed ethanol water, extracts in ammonium hydroxide that the precipitate adding is 0.15～0.2 mole and the butanol solution, and reject precipitate, distillation extraction liquid, concentrate carry out lyophilization and promptly obtain refining Venenum apis.
2, in accordance with the method for claim 1, it is characterized in that lixiviate is 6.4 with the PH of distilled water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100469560A CN101088514A (en) | 2006-06-16 | 2006-06-16 | Bee venom refining process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006100469560A CN101088514A (en) | 2006-06-16 | 2006-06-16 | Bee venom refining process |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101088514A true CN101088514A (en) | 2007-12-19 |
Family
ID=38942113
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2006100469560A Pending CN101088514A (en) | 2006-06-16 | 2006-06-16 | Bee venom refining process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101088514A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140314871A1 (en) * | 2012-01-04 | 2014-10-23 | Republic Of Korea (Management:Rural Development Administration) | Method for purifying bee venom on mass scale |
| US20150306025A1 (en) * | 2012-12-20 | 2015-10-29 | South China Sea Institute Of Oceanology, Chinese Academy Of Sciences | Bee Venom Composition with Effects of Protecting and Beautifying Lip |
| CN105596608A (en) * | 2014-10-17 | 2016-05-25 | 北京久颐蜂科技有限公司 | Bee venom essence |
| CN105878612A (en) * | 2014-10-17 | 2016-08-24 | 北京久颐蜂科技有限公司 | Bee venom essence and preparation method thereof |
| CN107056913A (en) * | 2017-03-21 | 2017-08-18 | 济南大学 | A kind of method for preparing melittin |
-
2006
- 2006-06-16 CN CNA2006100469560A patent/CN101088514A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140314871A1 (en) * | 2012-01-04 | 2014-10-23 | Republic Of Korea (Management:Rural Development Administration) | Method for purifying bee venom on mass scale |
| US9233129B2 (en) * | 2012-01-04 | 2016-01-12 | Republic Of Korea (Management: Rural Development Administration) | Method for purifying bee venom on mass scale |
| US20150306025A1 (en) * | 2012-12-20 | 2015-10-29 | South China Sea Institute Of Oceanology, Chinese Academy Of Sciences | Bee Venom Composition with Effects of Protecting and Beautifying Lip |
| US10085935B2 (en) * | 2012-12-20 | 2018-10-02 | South China Sea Institute Oceanology, Chinese Academy Of Sciences | Bee venom composition with effects of protecting and beautifying lip |
| CN105596608A (en) * | 2014-10-17 | 2016-05-25 | 北京久颐蜂科技有限公司 | Bee venom essence |
| CN105878612A (en) * | 2014-10-17 | 2016-08-24 | 北京久颐蜂科技有限公司 | Bee venom essence and preparation method thereof |
| CN107056913A (en) * | 2017-03-21 | 2017-08-18 | 济南大学 | A kind of method for preparing melittin |
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|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |