CN103059126A - Method for purifying menotrophins by ammonium acetate - Google Patents
Method for purifying menotrophins by ammonium acetate Download PDFInfo
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- CN103059126A CN103059126A CN2012105980026A CN201210598002A CN103059126A CN 103059126 A CN103059126 A CN 103059126A CN 2012105980026 A CN2012105980026 A CN 2012105980026A CN 201210598002 A CN201210598002 A CN 201210598002A CN 103059126 A CN103059126 A CN 103059126A
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- ammonium acetate
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Abstract
The invention discloses a method for purifying menotrophins by ammonium acetate. The method is characterized in that under the condition of a low temperature, a menotrophin crude product ( with the effective constituents lower than 10%) is extracted by an ammonium acetate solution, subjected to ethanol precipitation, and finally dehydrated with acetone, so that a menotrophin extract is obtained. In the technology, over 80% of impurities in the menotrophin crude product are removed; the menotrophin extract is adsorbed and purified through a resin column in subsequent processes, the usage amount of resin is saved, and the production cost is reduced; and hidden dangers that the body of the resin column is blocked, and even a danger of swelling and cracking of the resin column body appears due to the fact the menotrophin crude product has excessive impurities in an operation process are avoided.
Description
Technical field
The present invention relates to biological technical field, utilize Menotropins protein to be dissolved in characteristic in the finite concentration salts solution, adopt ammonium acetate solution purification Menotropins under the cold condition, the ethanol precipitation is at last with the acetone Menotropins purification thing that dewaters to get.
Background technology
Menotropins (HMG) is by the merocrine a kind of glycoprotein gonad-stimulating hormone of people's pituitary body frontal lobe, generally is to make through extracting purifying the urine of women after climacteric or climacteric.Women's content of HMG between menstrual period fluctuates to some extent, but lower, only have the women to enter in climacteric or more than ten years afterwards, (60-100iu/L urine) peaks the content side of HMG in its urine, and can maintain on the stable level, HMG contains follicular stimulating hormone (FSH) and lutropin (LH), approximate 1: 1 of its contained ratio.Clinically be mainly used in treating the women because of functional infertility due to not ovulating; The male sex is because of the low caused spermacrasia of hormonal readiness or vigor deficiency.In addition, also be used for the treatment of diseases such as women's amenorrhoea, menoxenia, the imbalance of men and women's sexual function.
Nineteen twenty-eight Ascbheim has at first reported from the climacteric women urine and has found active substance, though report is many afterwards, but to understanding and the preparation of HMG, be the Donimi since the sixties in 20th century really, and form gradually that more normalized technique prepares.China begins one's study from 20 century 70s, formally enters clinical experimental stage the eighties, begins a large amount of productions the nineties.
In actual production, because impurity component too much (effective constituent is less than 10%) in the Menotropins crude product if the Menotropins crude product can increase the resin usage quantity directly by the post adsorption and purification, because resin price is expensive, causes production cost to raise.And in the adsorption operations process, impurity easily stops up cylinder, even the danger of cylinder spalling may occur.The present invention is directed to these problems, technological method with ammonium acetate purification Menotropins crude product has been proposed, Menotropins after the purification has removed out 80% impurity, the usage quantity of in follow-up consonance resin absorption process, not only having saved resin, avoid the danger of cylinder spalling, also saved the time of whole operation.
Summary of the invention
The concrete grammar of the ammonium acetate purification Menotropins that the present invention proposes may further comprise the steps:
1) preparation of 5%-20% ammonium acetate solution: the 50g-200g ammonium acetate is dissolved in the ethanolic soln of 1000ml 70%, namely gets the ammonium acetate solution of 5%-20%.
2) with the Menotropins crude product in 1g-3g: the 15ml-35ml ratio adds in the ammonium acetate solution of 5%-20%, stirs 0.5-4.5 hour under 0-20 ℃ of condition, and is centrifugal, leaves and takes supernatant liquor.
3) centrifugal slag is in 1g-3g: the 15ml-30ml ratio adds the ammonium acetate solution of 5%-20%, stirs 0.5-4.5 hour under 0-20 ℃ of condition, and is centrifugal, leaves and takes supernatant liquor.
4) merge twice supernatant liquor, in 1ml-2ml: the 1.5ml-4.5ml ratio adds 95% ethanol, stirs 30-60 minute, sets low under warm 0-20 ℃ and leaves standstill 10-24 hour.
5) centrifugal, the collecting precipitation thing, centrifugal with the proper amount of acetone dehydration, throw out is equipped with Vanadium Pentoxide in FLAKES makes drying in the vacuum drying oven of water-retaining agent, get Menotropins purification thing.
The invention discloses the technological method of ammonium acetate purification Menotropins, its advantage and positively effect are that the Menotropins crude product is through ammonium acetate solution dissolving, ethanol precipitation, after the acetone dehydration, can effectively remove the impurity component in the Menotropins crude product, make it in the plastic resin treatment process, save resin demand, reduce production costs, can also avoid in addition the hidden danger that too much causes stopping up cylinder even cylinder spalling danger occurs because of Menotropins crude product impurity.
Specific embodiment
1) preparation of 7% ammonium acetate solution: the 70g ammonium acetate is dissolved in the ethanolic soln of 1000ml70%, namely gets 7% ammonium acetate solution.
2) 400g Menotropins crude product is added in 7% the ammonium acetate solution of 15L, under 10 ℃ of conditions, stirred 3 hours.Centrifugal, leave and take supernatant liquor.
3) 7% of centrifugal slag adding 8L ammonium acetate solution stirred 2 hours under 10 ℃ of conditions.Centrifugal, leave and take supernatant liquor.
4) merge twice supernatant liquor, add 95% ethanol of 40L, stirred 50 minutes, set low 10 ℃ of lower standing over night of temperature.
5) centrifugal, the collecting precipitation thing, centrifugal with the proper amount of acetone dehydration, throw out is equipped with Vanadium Pentoxide in FLAKES makes drying in the vacuum drying oven of water-retaining agent, get Menotropins purification thing 120g.
Advantage of the present invention and positively effect
120g Menotropins purification thing directly passes through the consonance resin absorption again through follow-up consonance resin absorption operation with 400g Menotropins crude product, has carried out the simultaneous test that operating time, cost, finished product are tired, and the results are shown in following table:
By relatively, sufficient proof the advantage of the present invention with respect to conventional art, not only slightly better aspect tiring, also removed impurity, saved the resin demand in the follow-up consonance operation, save the time of whole operation, thereby reduce production costs.
The above only is preferred embodiment of the present invention, is not to be the restriction of the present invention being made 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, foundation technical spirit of the present invention of not breaking away from still belongs to the protection domain of technical solution of the present invention to any simple modification, equivalent variations and remodeling that above embodiment does.
Claims (8)
1. the method for an ammonium acetate purification Menotropins, it is characterized in that, under cold condition, use ammonium acetate solution to extract the Menotropins crude product, afterwards the supernatant liquor that obtains used the ethanol precipitation, at last with the gained throw out with the acetone Menotropins purification thing that dewaters to get.
2. method according to claim 1 is characterized in that, may further comprise the steps:
(1) the Menotropins crude product is dropped in the ammonium acetate solution, under cold condition, stir, centrifugal, leave and take supernatant liquor;
(2) centrifugal slag adds ammonium acetate solution again, stirs under cold condition, and is centrifugal, leaves and takes supernatant liquor;
(3) merge twice supernatant liquor, add 95% ethanol, stir, set low under the warm environment, leave standstill;
(4) centrifugal, the collecting precipitation thing, centrifugal with the acetone dehydration, with throw out vacuum-drying, get Menotropins purification thing.
3. method according to claim 2 is characterized in that, the ratio of Menotropins crude product and ammonium acetate solution is 1g-3g: 15ml-35ml in the step (1); Cold condition is: 0-20 ℃; Churning time: 0.5-4.5 hour.
4. method according to claim 2 is characterized in that, the ratio of centrifugal slag and ammonium acetate extracting solution is 1g-3g: 15m1-30ml in the step (2); Cold condition: 0-20 ℃; Churning time: 0.5-4.5 hour.
5. method according to claim 2 is characterized in that, the volume ratio of supernatant liquor and 95% ethanol is 1-2: 1.5-4.5 in the step (3); Low temperature environment temperature: 0-20 ℃; Time of repose: 10-24 hour.
6. one of any described method is characterized in that according to claim 1-5, and described ammonium acetate solution is by preparing in the ethanolic soln that the 50g-200g ammonium acetate is dissolved in 1000ml70%.
7. one of any described method is characterized in that according to claim 1-6, and the Menotropins purification thing for preparing prepares the Menotropins finished product through the resin absorption operation again.
8. the Menotropins for preparing according to one of any described method of claim 1-7.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105980026A CN103059126A (en) | 2012-12-30 | 2012-12-30 | Method for purifying menotrophins by ammonium acetate |
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|---|---|---|---|
| CN2012105980026A CN103059126A (en) | 2012-12-30 | 2012-12-30 | Method for purifying menotrophins by ammonium acetate |
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| CN103059126A true CN103059126A (en) | 2013-04-24 |
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| CN2012105980026A Pending CN103059126A (en) | 2012-12-30 | 2012-12-30 | Method for purifying menotrophins by ammonium acetate |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497129A (en) * | 2014-12-23 | 2015-04-08 | 青岛康原药业有限公司 | Technology for purifying urofollitropin |
| CN105399813A (en) * | 2015-11-26 | 2016-03-16 | 青岛康原药业有限公司 | Method for extracting and purifying high-valence HMG (human menopausal gonadotropin) |
-
2012
- 2012-12-30 CN CN2012105980026A patent/CN103059126A/en active Pending
Non-Patent Citations (4)
| Title |
|---|
| 《生化药物杂志》 19901231 顾庆华 "人尿促性腺激素提取及部分纯化" 第46-47页 1-8 , 第2期 * |
| 《生化药物杂志》 19911231 顾庆华 "人尿促滤泡素和促黄体素分离提取" 第59-61页 1-8 , 第1期 * |
| 顾庆华: ""人尿促性腺激素提取及部分纯化"", 《生化药物杂志》 * |
| 顾庆华: ""人尿促滤泡素和促黄体素分离提取"", 《生化药物杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104497129A (en) * | 2014-12-23 | 2015-04-08 | 青岛康原药业有限公司 | Technology for purifying urofollitropin |
| CN105399813A (en) * | 2015-11-26 | 2016-03-16 | 青岛康原药业有限公司 | Method for extracting and purifying high-valence HMG (human menopausal gonadotropin) |
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Application publication date: 20130424 |