CN1712410A - Extraction of 4-androstenedione from oil phase of fermenting liquid - Google Patents
Extraction of 4-androstenedione from oil phase of fermenting liquid Download PDFInfo
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- CN1712410A CN1712410A CN 200410025163 CN200410025163A CN1712410A CN 1712410 A CN1712410 A CN 1712410A CN 200410025163 CN200410025163 CN 200410025163 CN 200410025163 A CN200410025163 A CN 200410025163A CN 1712410 A CN1712410 A CN 1712410A
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Abstract
Extraction of 4-androstenedione from oil phase of fermentation liquor achieves higher product purity, repeat use, no environmental pollution and short process route.
Description
Technical field:
The invention belongs to the vegetable chemistry technical field.Be specifically related to extract the oil phase (vegetables oil) from fermented liquid the method for 4-Androstenedione 4-Androstadiendione (4-AD).
Background technology:
Plant sterol is under the situation of media at oil phase (vegetables oil), after special fermentation using bacteria bio-transformation, plant sterol can be converted into 4-AD, because 4-AD has stronger lipophilicity, so mainly be present in the oil phase media in the fermented liquid.
Summary of the invention:
Technical problem to be solved by this invention is to overcome the weak point in the existing technology of extracting 4-AD, designs a kind of extract yield height, extracting method low in the pollution of the environment.
The invention provides the method for extracting 4-Androstenedione (4-AD) a kind of oil phase from fermented liquid (vegetables oil), this method comprises the following steps:
(1) oil phase of collection and extractive fermentation liquid:
In fermented liquid, add an amount of salt and make water and the layering of oil phase part in the fermented liquid, collection oil phase part, with the alcohol extraction of 1: 1 (V/V) 4-5 time, combining extraction liquid;
(2) extract 4-Androstenedione crude product I:
The extraction liquid of step (1) reduction vaporization under 45 ℃, 0.095Mpa condition is reclaimed ethanol, when small amount of crystal occurs, stops to concentrate, be cooled to 5 ℃ of crystallizations 8 hours, centrifugal collection crystal, crude product I;
(3) purifying makes the pure product of 4-Androstenedione:
With 1: 15 (W/V) acetone solution crude product I, filter, reduction vaporization reclaims acetone under 25 ℃, 0.095Mpa condition, stops to concentrate when small amount of crystal occurs, and is cooled to 5 ℃ of crystallizations 8 hours, centrifugal collection crystal, crude product II; With W/V ethyl acetate dissolving in 1: 20, add 0.1% W/V activated carbon stirring decolouring 1 hour, filter, reduction vaporization stops to concentrate when a small amount of crystallization is separated out, and is cooled to 5 ℃ of crystallizations 8 hours, the centrifugal crude product III that gets; With crude product III with 1: 25 W/V dissolve with methanol, add the 0.1%W/V activated carbon and stir decolouring 1 hour, filter, reduction vaporization to the time small amount of crystalline stop to concentrate when separating out, be cooled to 5 ℃ of crystallizations 8 hours, centrifugal, use the small amount of methanol washing crystal, 45 ℃, 0.095Mpa vacuum-drying 8 hours pure product 4-AD.
In addition, (V/V) ethyl acetate dissolving that the oil phase part of method steps of the present invention (1) behind alcohol extraction used 1: 2 again, standing demix 8 hours, supernatant liquor is handled back reduction vaporization under 45 ℃-60 ℃, 0.095Mpa condition after filtration and is reclaimed ethyl acetate, get clarifying refiltered oil, this oil can be recycled and reused for production.
Method of the present invention has following characteristics:
1. with the method for salt in fermented liquid, make fermented liquid water and oil phase layering, and make the 4-Androstenedione that is partially dissolved in aqueous portion all be dissolved in the oil phase;
2. be dissolved in 4-Androstenedione in the oil phase with the big organic solvent extraction of polarity, the 4-Androstenedione that is dissolved in the oil phase part is transferred in the extraction liquid;
3. use the solvent dissolution process crude product crystallization repeatedly of opposed polarity, make that the impurity of opposed polarity is effectively separated.Extraction liquid filters through concentrating, and cooling after time crude product crystallization of winning, with the crystallization of non-polar organic solvent dissolving crude product, is removed segment polarity impurity after filtering; Filtered liquid is through concentrating, cooling, after the crude product crystallization for the second time, with little this crude product crystallization of organic solvent dissolution of polarity, remove partial impurities after filtration; Filtered liquid is through concentrating, cooling, after the crude product crystallization for the third time, with the big organic solvent dissolution crude product crystallization of polarity, after decolorizing with activated carbon filters, remove part non polar impurities and pigment; Filtered liquid is through concentrating, and cooling gets pure product crystallization.
4. with the oil phase after the organic solvent dissolution processing extraction, make refiltered oil reach the purpose that to apply mechanically repeatedly.
Method of the present invention and existing technology relatively see Table 1:
Table 1 the technology of the present invention level and former technology level catalog data
| The technology of the present invention level | Former state of the art | ||||||
| Weight/kg | Product content % | Product is analysed pure % | Weight/kg | Product content % | Product is analysed pure % | ||
| The extraction level | Ferment oil * | ????12 | ????12 | ||||
| A crude product | ????20 | ????52.5 | ????10.5 | ????15.6 | ?????48 | ????7.5 | |
| The secondary crude product | ????15 | ????65.5 | ????9.8 | ????9.84 | ?????62 | ????6.1 | |
| Three crude products | ????11.75 | ????80 | ????9.4 | ????7.8 | ?????74.4 | ????5.8 | |
| Finished product | ????8.12 | ????98.5 | ????8 | ????5.86 | ?????95.6 | ????5.6 | |
| Final product quality | Fusing point | ??????????171.5～172.3℃ | ??????????????168.1～170.5℃ | ||||
| 1,4-Androstenedione content | ??????????0.16％ | ??????????????1.10％ | |||||
| Specific rotation | ??????????188.00％ | ??????????????180.00％ | |||||
| Yield (analysing pure) * | ??????????36.40％ | ??????????????25.50％ | |||||
| Purity | ??????????98.5 | ??????????????95.6％ | |||||
Method operational path of the present invention is short, product purity is high, refiltered oil can reuse, and reduces environmental pollution, suitable scale suitability for industrialized production.
*Analyse pure products in the ferment oil: meaning that plant sterol transfers to through fermentation is the total mass behind the AD.
*Yield (analysing pure): qualified product through analyse pure after with the ratio of the plant sterol weight that drops into, refer to weight yield.
Illustrate: above data are the average data of 5 experimental results.The present invention improves nearly 11 percentage points than former technology on extract yield as can be seen on the data.Product quality is from purity, fusing point, than the product that all is better than former technology on outstanding and the by product content.
Embodiment:
(1) 30kg salt is added in the 500L fermented liquid, stirred 30 minutes, static layering 8 hours is collected oil phase part 120L.The gradation extraction is 5 times in extractor, all uses the alcohol extraction of 1: 1 (V/V) at every turn, merges the about 610L of extraction liquid 5 times, (the raffinate about 110L of oil processing to be recycled).With 610L extraction liquid concentrating under reduced pressure in concentration tank, crystallisation by cooling, centrifuging gets the about 20kg of crude product; With 300L acetone solution 20kg crude product, filter, concentrating under reduced pressure in concentration tank, crystallisation by cooling, centrifuging gets the about 15kg of crude product; With 300L ethyl acetate dissolving 15kg crude product, add the 0.3kg activated carbon, stir decolouring 1 hour, filter, concentrating under reduced pressure in concentration tank, crystallisation by cooling, centrifuging gets the about 12kg of crude product; With 300L dissolve with methanol 12kg crude product, add the 0.3kg activated carbon, stir decolouring 1 hour, filter, concentrating under reduced pressure in concentration tank, crystallisation by cooling, the centrifuge washing crystallization, in vacuum drying oven 45 ℃, 0.095Mpa vacuum-drying 8 hours the about 8kg of pure product 4-AD;
(2) with the about 110L of raffinate oil 220L ethyl acetate stirring and dissolving, standing demix 8 hours discards the lower sediment part, supernatant liquor after filtering under 45 ℃-60 ℃, 0.095Mpa condition reduction vaporization reclaim ethyl acetate, get the about 80L of clarifying refiltered oil, this oil can be recycled and reused for production.
Claims (2)
1, a kind of method of extracting the 4-Androstenedione from plant sterol fermented liquid oil phase is characterized in that this method comprises the following steps:
(1) oil phase of collection and extractive fermentation liquid:
In fermented liquid, add an amount of salt and make water and the layering of oil phase part in the fermented liquid, collection oil phase part, with the alcohol extraction of 1: 1 (V/V) 4-5 time, combining extraction liquid;
(2) extract 4-Androstenedione crude product I:
The extraction liquid of step (1) reduction vaporization under 45 ℃, 0.095Mpa condition is reclaimed ethanol, when small amount of crystal occurs, stops to concentrate, be cooled to 5 ℃ of crystallizations 8 hours, centrifugal collection crystal, crude product I;
(3) purifying makes the pure product of 4-Androstenedione:
With 1: 15 (W/V) acetone solution crude product I, filter, reduction vaporization reclaims acetone under 25 ℃, 0.095Mpa condition, stops to concentrate when small amount of crystal occurs, and is cooled to 5 ℃ of crystallizations 8 hours, centrifugal collection crystal, crude product II; With 1: the dissolving of 20W/V ethyl acetate, add 0.1%W/V activated carbon stirring decolouring 1 hour, filter, reduction vaporization stops to concentrate when a small amount of crystallization is separated out, and is cooled to 5 ℃ of crystallizations 8 hours, the centrifugal crude product III that gets; With crude product III with 1: the 25W/V dissolve with methanol, add the 0.1%W/V activated carbon and stir decolouring 1 hour, filter, reduction vaporization to the time small amount of crystalline stop to concentrate when separating out, be cooled to 5 ℃ of crystallizations 8 hours, centrifugal, use the small amount of methanol washing crystal, 45 ℃, 0.095Mpa vacuum-drying 8 hours pure product 4-AD.
2, a kind of method of from plant sterol fermented liquid oil phase, extracting the 4-Androstenedione according to claim 1, it is characterized in that (V/V) ethyl acetate dissolving of using 1: 2 again of the oil phase part behind alcohol extraction of step (1) wherein, standing demix 8 hours, supernatant liquor is handled back reduction vaporization under 45 ℃-60 ℃, 0.095Mpa condition after filtration and is reclaimed ethyl acetate, get clarifying refiltered oil, this oil can be recycled and reused for production.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100251631A CN100457771C (en) | 2004-06-15 | 2004-06-15 | Extraction of 4-androstenedione from oil phase of fermenting liquid |
Applications Claiming Priority (1)
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|---|---|---|---|
| CNB2004100251631A CN100457771C (en) | 2004-06-15 | 2004-06-15 | Extraction of 4-androstenedione from oil phase of fermenting liquid |
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| Publication Number | Publication Date |
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| CN1712410A true CN1712410A (en) | 2005-12-28 |
| CN100457771C CN100457771C (en) | 2009-02-04 |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102432657A (en) * | 2011-11-03 | 2012-05-02 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting and purifying 4-androstenedione |
| CN102827913A (en) * | 2012-08-31 | 2012-12-19 | 湖南诺凯生物医药有限公司 | Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes |
| CN102936272A (en) * | 2012-03-27 | 2013-02-20 | 广东本科生物工程股份有限公司 | Process capable of circularly extracting AD and/or ADD from fermentation broth |
| CN103159816A (en) * | 2011-12-16 | 2013-06-19 | 山东方明药业集团股份有限公司 | Method of extracting 4- androstenedione from plant sterol fermentation liquor |
| CN103254261A (en) * | 2013-05-30 | 2013-08-21 | 浙江钱江生物化学股份有限公司 | Androstenedione purification method |
| CN103254262A (en) * | 2013-05-30 | 2013-08-21 | 浙江钱江生物化学股份有限公司 | Method for extracting androstenedione by adopting pipeline type continuous ultrasonic extraction technology |
| CN103360449A (en) * | 2013-07-24 | 2013-10-23 | 江苏久吾高科技股份有限公司 | Method for extracting androstenedione by use of membrane method |
| CN103613627A (en) * | 2013-11-20 | 2014-03-05 | 上海市农药研究所 | Method for recycling edible oil in androstenedione production process through biotransformation |
| CN103923144A (en) * | 2013-01-10 | 2014-07-16 | 上海医药工业研究院 | Androst-4-ene-3,17-dione polymorphism I, preparation method and application thereof |
| CN105175471A (en) * | 2014-06-06 | 2015-12-23 | 山东方明药业集团股份有限公司 | Method for extracting androstenedione crude product from two-liquid-phase system |
| CN110343139A (en) * | 2018-05-24 | 2019-10-18 | 石家庄学院 | The method of 4-AD is extracted from 4-AD industry waste liquor |
| CN112175033A (en) * | 2020-10-30 | 2021-01-05 | 湖南新合新生物医药有限公司 | Method for refining steroid intermediate mother liquor material |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB1092145A (en) * | 1964-06-02 | 1967-11-22 | Noda Inst For Scientific Res | Microbiological conversion of steroids |
| US4293644A (en) * | 1976-10-22 | 1981-10-06 | The Upjohn Company | Process for preparing androst-4-ene-3,17-dione |
| JPH06153987A (en) * | 1992-11-17 | 1994-06-03 | Nippon Kayaku Co Ltd | New production of 14alpha-hydroxy-4-androstene-3,17-dione |
| KR100432054B1 (en) * | 2001-05-11 | 2004-05-17 | (주)유진사이언스 | Microorganism having ability to convert sterol into Androst-4-ene-3,17-dione/Androsta-1,4-diene-3,17-dione and preparation method thereof |
| CA2474575A1 (en) * | 2002-02-01 | 2003-08-07 | Edward J. Herrington | Process for fermentation of phytosterols to androstadienedione |
-
2004
- 2004-06-15 CN CNB2004100251631A patent/CN100457771C/en not_active Expired - Fee Related
Cited By (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102432657A (en) * | 2011-11-03 | 2012-05-02 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting and purifying 4-androstenedione |
| CN103159816A (en) * | 2011-12-16 | 2013-06-19 | 山东方明药业集团股份有限公司 | Method of extracting 4- androstenedione from plant sterol fermentation liquor |
| CN103159816B (en) * | 2011-12-16 | 2015-09-30 | 山东方明药业集团股份有限公司 | A kind of method extracting 4-AD from phytosterol fermentation broth |
| CN102936272A (en) * | 2012-03-27 | 2013-02-20 | 广东本科生物工程股份有限公司 | Process capable of circularly extracting AD and/or ADD from fermentation broth |
| CN102936272B (en) * | 2012-03-27 | 2015-08-05 | 广东本科生物工程股份有限公司 | The technique of AD and/or ADD in a kind of extraction fermented liquid capable of circulation |
| CN102827913A (en) * | 2012-08-31 | 2012-12-19 | 湖南诺凯生物医药有限公司 | Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes |
| CN102827913B (en) * | 2012-08-31 | 2014-01-15 | 湖南成大生物科技有限公司 | Method for preparing 11 alpha-OH-ADD by mixed fermentation of microbes |
| CN103923144A (en) * | 2013-01-10 | 2014-07-16 | 上海医药工业研究院 | Androst-4-ene-3,17-dione polymorphism I, preparation method and application thereof |
| CN103254261A (en) * | 2013-05-30 | 2013-08-21 | 浙江钱江生物化学股份有限公司 | Androstenedione purification method |
| CN103254262A (en) * | 2013-05-30 | 2013-08-21 | 浙江钱江生物化学股份有限公司 | Method for extracting androstenedione by adopting pipeline type continuous ultrasonic extraction technology |
| CN103360449B (en) * | 2013-07-24 | 2015-03-25 | 江苏久吾高科技股份有限公司 | Method for extracting androstenedione by use of membrane method |
| CN103360449A (en) * | 2013-07-24 | 2013-10-23 | 江苏久吾高科技股份有限公司 | Method for extracting androstenedione by use of membrane method |
| CN103613627A (en) * | 2013-11-20 | 2014-03-05 | 上海市农药研究所 | Method for recycling edible oil in androstenedione production process through biotransformation |
| CN103613627B (en) * | 2013-11-20 | 2017-12-01 | 上海市农药研究所 | The recycling method of food-grade oil during bioconversion production androstenedione |
| CN105175471A (en) * | 2014-06-06 | 2015-12-23 | 山东方明药业集团股份有限公司 | Method for extracting androstenedione crude product from two-liquid-phase system |
| CN110343139A (en) * | 2018-05-24 | 2019-10-18 | 石家庄学院 | The method of 4-AD is extracted from 4-AD industry waste liquor |
| CN110343139B (en) * | 2018-05-24 | 2020-07-17 | 石家庄学院 | Method for extracting 4-androstenedione from industrial waste mother liquor of 4-androstenedione |
| CN112175033A (en) * | 2020-10-30 | 2021-01-05 | 湖南新合新生物医药有限公司 | Method for refining steroid intermediate mother liquor material |
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| Publication number | Publication date |
|---|---|
| CN100457771C (en) | 2009-02-04 |
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