CN103665145A - Salting-in method for purification of chorionic gonadotrophin - Google Patents
Salting-in method for purification of chorionic gonadotrophin Download PDFInfo
- Publication number
- CN103665145A CN103665145A CN201310648004.6A CN201310648004A CN103665145A CN 103665145 A CN103665145 A CN 103665145A CN 201310648004 A CN201310648004 A CN 201310648004A CN 103665145 A CN103665145 A CN 103665145A
- Authority
- CN
- China
- Prior art keywords
- chorionic gonadotrophin
- high salt
- ethanol
- ratio
- supernatant liquor
- 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
- 229960004407 chorionic gonadotrophin Drugs 0.000 title claims abstract description 18
- 238000000034 method Methods 0.000 title claims abstract description 15
- 238000000746 purification Methods 0.000 title claims abstract description 7
- 150000003839 salts Chemical class 0.000 claims abstract description 30
- 238000003756 stirring Methods 0.000 claims abstract description 19
- 238000000605 extraction Methods 0.000 claims abstract description 16
- 239000000047 product Substances 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 10
- 239000012043 crude product Substances 0.000 claims abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- 239000006228 supernatant Substances 0.000 claims description 14
- 239000007788 liquid Substances 0.000 claims description 10
- 238000001291 vacuum drying Methods 0.000 claims description 5
- 239000002244 precipitate Substances 0.000 claims description 4
- 239000012535 impurity Substances 0.000 abstract description 2
- 230000003993 interaction Effects 0.000 abstract description 2
- 238000001035 drying Methods 0.000 abstract 2
- 238000012869 ethanol precipitation Methods 0.000 abstract 2
- 239000012530 fluid Substances 0.000 abstract 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 2
- 239000012362 glacial acetic acid Substances 0.000 description 2
- 229940088597 hormone Drugs 0.000 description 2
- 239000005556 hormone Substances 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 229960004249 sodium acetate Drugs 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- DPDMMXDBJGCCQC-UHFFFAOYSA-N [Na].[Cl] Chemical compound [Na].[Cl] DPDMMXDBJGCCQC-UHFFFAOYSA-N 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 239000003163 gonadal steroid hormone Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000000938 luteal effect Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001817 pituitary effect Effects 0.000 description 1
- 230000003169 placental effect Effects 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000002381 testicular Effects 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 210000002993 trophoblast Anatomy 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Reproductive Health (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a salting-in process for purification of chorionic gonadotrophin. The process is as below: adding a prepared high salt extract into a crude product solution, stirring fully, standing for overnight, and carrying out ethanol precipitation and drying to obtain a high salt extraction powder; adding a low salt reverse extraction fluid to the high salt extraction powder; and carrying out ethanol precipitation and drying to obtain a chorionic gonadotrophin intermediate. The process through the interaction of high-low salt reverse extraction effectively removes impurities in the chorionic gonadotrophin crude product while better guaranteeing titer of the product, so as to greatly improve the quality of the product.
Description
Technical field
The present invention relates to biological technical field, utilize the interaction of high-less salt back suction, the impurity in chorionic gonadotrophin crude product is removed better, reach the effect that improves product purity.
Background technology
Chorionic gonadotrophin is gonad-stimulating hormone medicine.Be from zygote implantation the 1st day, by placental trophoblast syncytium, secreted, in the time of 60-70 days, peak.After this decline gradually, about minute 4 days puerperiums, disappear.To women, luteal function can be promoted and maintain, Folliculogenesis and maturation can be promoted.To the male sex, can make pituitary function deficiency person's testis produce male sex hormone, impel the growth of testicular descent and male secondary sex characters.
First nineteen twenty-seven Zondek and Ascbheim find that gravid woman exists gonad-stimulating hormone in urinating, and Brown in 1936 and Venning find at pregnant 60-80 days, and in urine, chorionic gonadotrophin secretion reaches climax.
Our company, when purification chorionic gonadotrophin crude product, adds high salt, low salts solution by priority, under low temperature environment, chorionic gonadotrophin crude product is further purified to intermediate.In this process, by controlling the pH value of crude product solution, under slant acidity environment, reduce temperature simultaneously, height salts solution can fully be purified chorionic gonadotrophin crude product solution, reach the object that chorionic gonadotrophin crude product is purified, thereby obtain the higher intermediate of tiring.This method is compared with general method of purification, not only can obtain the intermediate of high-titer, and does not affect the yield of chorionic gonadotrophin, and the removal of high less salt is also very convenient, greatly reduces cost.
The concrete grammar of the molten method purification of the salt chorionic gonadotrophin that the present invention proposes, comprises the following steps:
1, the preparation of high salt extraction liquid: 0.1mol/L sodium-acetate, 0.015mol/L Calcium Chloride Powder Anhydrous, 0.5mol/L sodium-chlor, adds purified water stirring and dissolving, drips Glacial acetic acid, adjusts pH to 4.0-5.1; Then add ethanol, making ethanol final concentration is 50%, is uniformly mixed, and puts 5-19 ℃ of preservation.
2, by raw product 1:(16-23 by weight) ratio is slowly added to high salt extraction liquid, stirs 5 hours standing over night under solution temperature 5-19 ℃ condition.
3,1:(1.3-4.2 by volume in supernatant liquor) ratio adds-10 ℃ of following ethanol, adds, and continue to stir 30 minutes while stirring, and sets low warm 5-19 ℃ standing over night.
4, centrifugal in whizzer, abandoning supernatant, throw out is put the dry behaviour of vacuum in vacuum drying oven, the dried high salt extraction dry powder that is.
5, the preparation of less salt back suction liquid: 0.1mol/L sodium-acetate adds purified water stirring and dissolving, drips Glacial acetic acid, adjusts pH to 3.9-5.0; Then add ethanol, making ethanol final concentration is 50%, is uniformly mixed, and puts 5-19 ℃ of preservation.
6, by high salt extraction dry powder by weight 1:24-33 ratio slowly add less salt back suction liquid, under solution temperature 5-19 ℃ condition, stir 3 hours, centrifugal.
7, in supernatant liquor, by volume 1:1.3-4.2 ratio adds-10 ℃ of following ethanol, adds while stirring, and continues to stir 30 minutes, sets low warm 5-19 ℃ standing over night.
8, centrifugal in whizzer, abandoning supernatant, throw out is put the dry behaviour of vacuum in vacuum drying oven, the dried middle product of chorionic gonadotrophin that are.
Specific embodiment
Embodiment 1
(1) get raw product 700g, add the high salt extraction liquid of 14LpH5.1, under 10 ℃ of conditions of low temperature, stir 5 hours standing over night;
(2) centrifuging and taking supernatant liquor, adds the ratio of 3L ethanol in 1L supernatant liquor, add in advance while stirring 95% ethanol that is chilled to-10 ℃, and continues to stir 30 minutes, sets low 10 ℃ of standing over night of temperature;
(3) precipitate centrifugal collection, abandoning supernatant, throw out is put the dry behaviour of vacuum in vacuum drying oven, after being dried, is high salt extraction dry powder, and 50.4g weighs.
(4) get high salt extraction dry powder, slowly add the less salt back suction liquid of 1.5LpH4.8, under 10 ℃ of conditions of low temperature, stir 3 hours, centrifugal, collect supernatant liquor;
(5) in supernatant liquor, add while stirring the ethanol that 6L is chilled to-10 ℃ in advance, and continue to stir 30 minutes, set low 10 ℃ of standing over night of temperature;
(6) precipitate centrifugal collection, abandoning supernatant, throw out is put in vacuum drying oven the dry behaviour of vacuum, the product that are after dry in the middle of the chorionic gonadotrophin 37.8g that weighs.Be that 700g raw product can obtain the middle product of 37.8g.
The above, be only preferred embodiment of the present invention, is not the present invention to be done to the restriction of other form, and any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the equivalent embodiment of equivalent variations.But every technical solution of the present invention content that do not depart from, any simple modification, equivalent variations and the remodeling above embodiment done according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
Claims (9)
1. with the molten method purification of salt chorionic gonadotrophin, comprise the following steps:
(1) raw product is added to high salt extraction liquid, after reaching certain temperature, stirs standing over night;
(2) in supernatant liquor, add ethanol, while stirring, add, standing over night, precipitates centrifugal collection, after being dried, is high salt extraction dry powder;
(3) high salt extraction dry powder is added to less salt back suction liquid, after reaching certain temperature, stir, centrifugal;
(4) get supernatant liquor, add ethanol, add while stirring, standing over night, precipitates centrifugal collection, and vacuum-drying obtains the middle product of chorionic gonadotrophin.
2. method according to claim 1, is characterized in that, in step (1), high salt extraction liquid adds in the ratio of the 1:16-23 of crude product weight.
3. method according to claim 1, is characterized in that, in step (1), solution temperature is 5-19 ℃.
4. method according to claim 1, is characterized in that, in step (2), in the ratio of the volume ratio 1:1.3-4.2 of supernatant liquor, adds ethanol.
5. according to described in claim 1 and 4, it is characterized in that, ethanol temperature is controlled at below-10 ℃.
6. method according to claim 1, is characterized in that, in step (3), less salt back suction liquid adds in the ratio of the 1:24-33 of high salt extraction dry powder weight.
7. method according to claim 1, is characterized in that, in step (3), solution temperature is 5-19 ℃.
8. method according to claim 1, is characterized in that, in step (4), in the ratio of the volume ratio 1:1.3-4.2 of supernatant liquor, adds ethanol.
9. according to described in claim 1 and 8, it is characterized in that, ethanol temperature is controlled at below-10 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310648004.6A CN103665145A (en) | 2013-11-27 | 2013-11-27 | Salting-in method for purification of chorionic gonadotrophin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310648004.6A CN103665145A (en) | 2013-11-27 | 2013-11-27 | Salting-in method for purification of chorionic gonadotrophin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103665145A true CN103665145A (en) | 2014-03-26 |
Family
ID=50304005
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310648004.6A Pending CN103665145A (en) | 2013-11-27 | 2013-11-27 | Salting-in method for purification of chorionic gonadotrophin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103665145A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497128A (en) * | 2014-12-23 | 2015-04-08 | 青岛康原药业有限公司 | Method for extracting chorionic gonadotrophin from crude product via low-salt extraction method |
| CN105330734A (en) * | 2015-11-13 | 2016-02-17 | 山东众山生物科技有限公司 | Menotrophin purifying method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302818A (en) * | 2000-12-12 | 2001-07-11 | 上海惠海生化制品厂 | Human chorionic gonadotropin and its preparing process |
-
2013
- 2013-11-27 CN CN201310648004.6A patent/CN103665145A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1302818A (en) * | 2000-12-12 | 2001-07-11 | 上海惠海生化制品厂 | Human chorionic gonadotropin and its preparing process |
Non-Patent Citations (2)
| Title |
|---|
| 张日华: "高效价人绒毛膜促性腺激素的制备新方法", 《中国药科大学学报》 * |
| 徐桢琦: "人绒毛膜促性腺激素的精制、去热原工艺", 《沈阳药科大学学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497128A (en) * | 2014-12-23 | 2015-04-08 | 青岛康原药业有限公司 | Method for extracting chorionic gonadotrophin from crude product via low-salt extraction method |
| CN105330734A (en) * | 2015-11-13 | 2016-02-17 | 山东众山生物科技有限公司 | Menotrophin purifying method |
| CN105330734B (en) * | 2015-11-13 | 2018-11-23 | 山东众山生物科技有限公司 | A kind of method of purification of menotropins |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102885855B (en) | Production method for preparing medicine for treating peptic ulcer by periplaneta americana extract | |
| CN102038254A (en) | Chrysanthemum drink and preparation method thereof | |
| CN102432629A (en) | Method for refining ceftriaxone sodium crude product | |
| CN103665145A (en) | Salting-in method for purification of chorionic gonadotrophin | |
| CN103833052A (en) | Production method of food-grade potassium chloride | |
| CN101456822A (en) | Novel process for extracting threonine | |
| CN102225973A (en) | Production method for refined heparin sodium | |
| CN103193880B (en) | Preparation method of HCG (Human Chorionic Gonadotropin) crude product | |
| CN104095176A (en) | Honey clarifying method | |
| CN103553901B (en) | Preparation method of soluble calcium citrate | |
| CN103734365A (en) | Manufacturing process for dual-calcium-fortified low-fluorine brick tea | |
| CN102796189A (en) | Preparation method of protamine sulfate | |
| CN102898546A (en) | Method for extracting low-potency heparin sodium | |
| CN103666964B (en) | A kind of kidney healthful tonic wine and preparation method thereof | |
| CN105801687A (en) | Improved medicinal high-purity insulin extraction technology | |
| CN104530216A (en) | Method for extracting chorionic gonadotrophin from crude product by virtue of high-concentration salt solution extraction method | |
| CN106589108A (en) | Method for improving stability of chorionic gonadotrophin | |
| CN105085660A (en) | Method for extracting chorionic gonadotrophin crude product from large-volume pregnant woman urine on basis of ultrafiltration concentration | |
| CN102898492B (en) | Technique for extracting cholesterol from Antarctic krill | |
| CN103739700A (en) | Method using sodium benzoate for extraction of chorionic gonadotrophin | |
| CN108142829A (en) | A kind of production method of malt selenium | |
| CN101550144A (en) | Preparation technique for mezlocillin | |
| CN102302768A (en) | Method for preparing brain protein hydrolysate oral liquid | |
| CN104497128A (en) | Method for extracting chorionic gonadotrophin from crude product via low-salt extraction method | |
| CN103012581A (en) | Albumin preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140326 |