CN105439847A - Separation purification method for natural abscisic acid - Google Patents
Separation purification method for natural abscisic acid Download PDFInfo
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- CN105439847A CN105439847A CN201510972969.XA CN201510972969A CN105439847A CN 105439847 A CN105439847 A CN 105439847A CN 201510972969 A CN201510972969 A CN 201510972969A CN 105439847 A CN105439847 A CN 105439847A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/43—Separation; Purification; Stabilisation; Use of additives by change of the physical state, e.g. crystallisation
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C51/00—Preparation of carboxylic acids or their salts, halides or anhydrides
- C07C51/42—Separation; Purification; Stabilisation; Use of additives
- C07C51/47—Separation; Purification; Stabilisation; Use of additives by solid-liquid treatment; by chemisorption
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Abstract
A separation purification method for natural abscisic acid belongs to the field of biological fermentation and production of biochemical pesticides. The method comprises performing plate-frame filtration on an abscisic acid fermentation liquid to obtain a plate-frame filtrate, then processing the filtrate through an ultrafiltration membrane and a condensation membrane, so as to obtain a concentrate, acidifying the condensate, then performing extraction, pretreating the extraction liquid, then adding an eluent, adsorbing impurities through adsorption column chromatography, performing reduced-pressure concentration on the chromatography liquid, adding a non-polar solvent into the concentrate, filtering and drying, so as to obtain the abscisic acid product. The method is simple and convenient, abscisic acid extracted from the fermentation liquid is high in purity and high in yield, and product residue in the mother liquid is less.
Description
Technical field
The present invention relates to biological fermentation---biochemical pesticides production field, specifically belongs to a kind of separation purification method of PBI 58.
Background technology
Dormin is a kind of Endogenous hormones with comprehensive physiological function, one of five large plant hormones, it is the important growth regulator of plant, it is the key factor of balance plant endogenous hormones and the metabolism of related growth active substance, there is the ability promoting plant Balance Absorption water, fertilizer and coordinate internal metabolism, can the root/be preced with and nourish and grow and reproductive growth of Effective Regulation plant, to quality, the output improving farm crop, there is vital role.Dormin is an extraordinary product of market outlook, and agriculture production has broad application prospects, and can produce huge economic benefit and social benefit.Current dormin price is commercially still very high, 2005,98% PBI 58 (article number A4906-1MG), 1 milligram of price of Sigma Co., USA is Renminbi 3600 yuan, and be no matter at home or world market, dormin product all will be in the situation that supply falls short of demand for a long time.
At present, conventional dormin production method is biological fermentation process, this method reduce the production cost of PBI 58, but there is the low defect of yield, how high-level efficiency, extracting from fermented liquid of low cost obtain highly purified PBI 58 and become research emphasis.
Summary of the invention
The present invention is directed to the deficiency that current PBI 58 purification efficiency is low, cost is high, provide a kind of yield high, the separation purification method of the PBI 58 that cost is low.
Concrete technical scheme of the present invention is as follows.
A separation purification method for PBI 58, comprises following steps successively:
(1) dormin fermentation liquor Plate Filtration obtains sheet frame filtrate;
(2) sheet frame filtrate ultrafiltration membrane treatment, obtains ultrafiltration and concentration liquid and ultrafiltration dialysis liquid;
(3) ultrafiltration dialysis liquid is concentrated by concentrated film, obtains membrane concentration liquid;
(4) membrane concentration liquid extracts after acidifying, and extraction liquid adds eluent after pretreatment, after adsorpting column chromatography adsorbing contaminant, obtain chromatographic solution;
(5) chromatographic solution concentrating under reduced pressure, crystallisation by cooling, filtration, drying obtain dormin product.
In step (1), sheet frame filtrate pH is adjusted to 4.8-7.0.
In step (2), the molecular weight cut-off of ultra-filtration membrane is 3000-30000.
In step (3), concentrated film is reverse osmosis membrane or nanofiltration membrane or both combinations.
In step (3), the molecular weight cut-off of concentrated film is 50-200.
In step (4), the extraction agent of extraction is one or more in ethyl ester class, butyl ester class, isopentyl ester class.Extraction liquid pre-treatment is 60-120mg/ml for extraction liquid is concentrated into concentration.Eluent is preferably one or more in chloroform, methylene dichloride, sherwood oil, normal hexane.
In step (4), the sorbent material of adsorpting column chromatography can be one or more in aluminum oxide, silica gel, Magnesium Silicate q-agent.
Concentrating under reduced pressure described in step (5), its vacuum tightness can be preferably-0.08 ~-0.095Mpa, and chuck circulating water temperature can be preferably 45 ~ 60 DEG C, and material temperature can be preferably 30 ~ 40 DEG C.
In step (5), chromatographic solution is through concentrating under reduced pressure, promoting that mother liquor separates out powder again, increasing a productive rate of dormin product in concentrated solution by adding non-polar solvent.Non-polar solvent is preferably hexane, sherwood oil, trichloromethane, tetracol phenixin, and its ratio is 0.7 ~ 1.0 times of chromatography concentrated solution volume.
Beneficial effect of the present invention is: method of the present invention is simple, convenient, extracts that dormin purity is high, yield is high from fermented liquid, and in mother liquor, product residue is few.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
embodiment 1.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 4.9, be the ultrafiltration membrane treatment of 3000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, the nanofiltration membrane that ultrafiltration dialysis liquid is 100 by molecular weight cut-off concentrates, obtain concentrated solution, concentrated solution adds ethyl ester extraction after acidifying, extraction liquid is concentrated into 60mg/ml, then methylene dichloride is added, through silica gel column chromatography adsorbing contaminant, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.084Mpa, chuck circulating water temperature is 60 DEG C, material temperature is 40 DEG C, crystallisation by cooling, filter, drying obtains the dormin product that content is 96%.
embodiment 2.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 5.5, be the ultrafiltration membrane treatment of 5000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, the nanofiltration membrane that ultrafiltration dialysis liquid is 150 by molecular weight cut-off concentrates, obtain concentrated solution, concentrated solution adds butyl ester extraction after acidifying, extraction liquid is concentrated into 70mg/ml, then sherwood oil is added, through alumina column chromatography adsorbing contaminant, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.09Mpa, chuck circulating water temperature is 58 DEG C, material temperature is 38 DEG C, the hexane of 0.8 times is added after concentrated end, crystallisation by cooling, filter, drying obtains the dormin product that content is 95.2%.
embodiment 3.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 6.0, be the ultrafiltration membrane treatment of 10000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, the reverse osmosis membrane that ultrafiltration dialysis liquid is 50 by molecular weight cut-off concentrates, obtain concentrated solution, concentrated solution adds Isoamyl Acetate FCC extraction after acidifying, extraction liquid is concentrated into 80mg/ml, then chloroform is added, through Magnesium Silicate q-agent Adsorption impurity, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.092Mpa, chuck circulating water temperature is 45 DEG C, material temperature is 30 DEG C, the trichloromethane of 0.7 times is added in concentration process, crystallisation by cooling, filter, drying obtains the dormin product that content is 97%.
embodiment 4.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 6.5, be the ultrafiltration membrane treatment of 30000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, ultrafiltration dialysis liquid by molecular weight cut-off be 50 reverse osmosis membrane and molecular weight be 200 nanofiltration membrane concentrate, obtain concentrated solution, nanofiltration concentrated solution adds Isoamyl Acetate FCC extraction after acidifying, extraction liquid is concentrated into 100mg/ml, then normal hexane is added, through Magnesium Silicate q-agent Adsorption impurity, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.093Mpa, chuck circulating water temperature is 50 DEG C, material temperature is 33 DEG C, crystallisation by cooling, after filtering, filtrate adds the tetracol phenixin of 0.9 times, combination drying obtains the dormin product that content is 95%.
embodiment 5.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 7.0, be the ultrafiltration membrane treatment of 10000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, ultrafiltration dialysis liquid by molecular weight cut-off be 50 reverse osmosis membrane and molecular weight be 150 nanofiltration membrane concentrate, obtain concentrated solution, nanofiltration concentrated solution adds butyl ester extraction after acidifying, extraction liquid is concentrated into 90mg/ml, then methylene dichloride is added, through silica gel column chromatography adsorbing contaminant, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.088Mpa, chuck circulating water temperature is 54 DEG C, material temperature is 34 DEG C, after concentrated, filtrate adds the sherwood oil of 0.8 times, crystallisation by cooling, combination drying obtains the dormin product that content is 97.5%.
embodiment 6.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 5.5, be the ultrafiltration membrane treatment of 10000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, the nanofiltration membrane that ultrafiltration dialysis liquid is 150 by molecular weight cut-off concentrates, obtain concentrated solution, nanofiltration concentrated solution adds ethyl ester extraction after acidifying, extraction liquid is concentrated into 110mg/ml, then sherwood oil is added, through alumina column chromatography adsorbing contaminant, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.09Mpa, chuck circulating water temperature is 55 DEG C, material temperature is 35 DEG C, crystallisation by cooling, after filtering, filtrate adds the sherwood oil of 0.7 times, combination drying obtains the dormin product that content is 95.8%.
embodiment 7.
Dormin fermentation liquor Plate Filtration obtains sheet frame filtrate, pH is adjusted to 4.8, be the ultrafiltration membrane treatment of 10000 with molecular weight cut-off, obtain ultrafiltration dialysis liquid, ultrafiltration dialysis liquid by molecular weight cut-off be 50 reverse osmosis membrane and molecular weight be 150 nanofiltration membrane concentrate, obtain concentrated solution, nanofiltration concentrated solution adds ethyl ester extraction after acidifying, extraction liquid is concentrated into 100mg/ml, then chloroform is added, through silica gel column chromatography adsorbing contaminant, the chromatographic solution concentrating under reduced pressure obtained, vacuum tightness is-0.092Mpa, chuck circulating water temperature is 48 DEG C, material temperature is 32 DEG C, crystallisation by cooling, drying obtains the dormin product that content is 95%.
Claims (10)
1. a separation purification method for PBI 58, is characterized in that: comprise following steps successively:
(1) dormin fermentation liquor Plate Filtration obtains sheet frame filtrate;
(2) sheet frame filtrate ultrafiltration membrane treatment, obtains ultrafiltration and concentration liquid and dialyzate;
(3) ultrafiltration dialysis liquid is concentrated by concentrated film, obtains membrane concentration liquid;
(4) membrane concentration liquid extracts after acidifying, and extraction liquid adds eluent after pretreatment, after adsorpting column chromatography adsorbing contaminant, obtain chromatographic solution;
(5) chromatographic solution concentrating under reduced pressure, crystallisation by cooling, filtration, drying obtain dormin product.
2. the separation purification method of a kind of PBI 58 according to claim 1, is characterized in that in step (1), sheet frame filtrate pH is adjusted to 4.8 ~ 7.0.
3. the separation purification method of a kind of PBI 58 according to claim 1, is characterized in that the molecular weight cut-off of ultra-filtration membrane in step (2) is 3000 ~ 30000.
4. the separation purification method of a kind of PBI 58 according to claim 1, to it is characterized in that in step (3) that concentrated film is reverse osmosis membrane or nanofiltration membrane or both combinations, molecular weight cut-off is 50 ~ 200.
5. the separation purification method of a kind of PBI 58 according to claim 1, it is characterized in that extraction adopts in step (4) extraction agent is one or more in ethyl ester class, butyl ester class, isopentyl ester class, in step (4), extraction liquid pre-treatment is for be concentrated into 60 ~ 120mg/ml by extraction liquid.
6. the separation purification method of a kind of PBI 58 according to claim 1, is characterized in that the sorbent material of adsorpting column chromatography in step (4) is one or more in aluminum oxide, silica gel, Magnesium Silicate q-agent.
7. the separation purification method of a kind of PBI 58 according to claim 1, is characterized in that eluent in step (4) is one or more in chloroform, methylene dichloride, sherwood oil, normal hexane.
8. the separation purification method of a kind of PBI 58 according to claim 1, is characterized in that the concentrating under reduced pressure described in step (5), and its vacuum tightness is-0.08 ~-0.095Mpa, and chuck circulating water temperature is 45 ~ 60 DEG C, and material temperature is 30 ~ 40 DEG C.
9. according to the separation purification method of any one PBI 58 described in claim 1 ~ 8, it is characterized in that in step (5), chromatographic solution is through concentrating under reduced pressure, non-polar solvent is added in concentrated solution, the adding procedure of non-polar solvent is in chromatographic solution concentration process or the concentrated end of chromatographic solution is rear or after filtering, the adding proportion of non-polar solvent is 0.7 ~ 1.0 times of chromatography concentrated solution volume.
10. the separation purification method of a kind of PBI 58 according to claim 9, is characterized in that described non-polar solvent is one or more in hexane, sherwood oil, trichloromethane, tetracol phenixin.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106905142A (en) * | 2017-04-18 | 2017-06-30 | 四川龙蟒福生科技有限责任公司 | A kind of isolation and purification method of S abscisic acids |
| CN107047552A (en) * | 2017-04-18 | 2017-08-18 | 四川龙蟒福生科技有限责任公司 | The recoverying and utilizing method of accessory substance in a kind of S abscisic acids production |
| WO2018192529A1 (en) * | 2017-04-18 | 2018-10-25 | 四川龙蟒福生科技有限责任公司 | Method for preparing s-abscisic acid |
| CN116730831A (en) * | 2023-07-03 | 2023-09-12 | 江西新瑞丰生化股份有限公司 | Extraction method of abscisic acid mother liquor |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1254508A (en) * | 1998-11-21 | 2000-05-31 | 昆明恒溢隆经贸有限责任公司 | Preparation method of natural bud inhibitor and application of natural bud inhibitor in axillary bud inhibition of flue-cured tobacco |
| CN1749401A (en) * | 2004-09-14 | 2006-03-22 | 中国科学院成都生物研究所 | A kind of extracting method of PBI 58 |
| CN1944385A (en) * | 2005-10-08 | 2007-04-11 | 中国科学院成都生物研究所 | Method for separating and extracting abscisic acid from fermented liquid by ionic exchanging and reversed phase chromatography |
| CN103274927A (en) * | 2013-06-08 | 2013-09-04 | 南京化工职业技术学院 | Purification method for nature abscisic acid |
| CN104817452A (en) * | 2014-12-19 | 2015-08-05 | 三达膜科技（厦门）有限公司 | Method for extracting abscisic acid from abscisic acid fermentation liquor |
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2015
- 2015-12-23 CN CN201510972969.XA patent/CN105439847A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1254508A (en) * | 1998-11-21 | 2000-05-31 | 昆明恒溢隆经贸有限责任公司 | Preparation method of natural bud inhibitor and application of natural bud inhibitor in axillary bud inhibition of flue-cured tobacco |
| CN1749401A (en) * | 2004-09-14 | 2006-03-22 | 中国科学院成都生物研究所 | A kind of extracting method of PBI 58 |
| CN1944385A (en) * | 2005-10-08 | 2007-04-11 | 中国科学院成都生物研究所 | Method for separating and extracting abscisic acid from fermented liquid by ionic exchanging and reversed phase chromatography |
| CN103274927A (en) * | 2013-06-08 | 2013-09-04 | 南京化工职业技术学院 | Purification method for nature abscisic acid |
| CN104817452A (en) * | 2014-12-19 | 2015-08-05 | 三达膜科技（厦门）有限公司 | Method for extracting abscisic acid from abscisic acid fermentation liquor |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106905142A (en) * | 2017-04-18 | 2017-06-30 | 四川龙蟒福生科技有限责任公司 | A kind of isolation and purification method of S abscisic acids |
| CN107047552A (en) * | 2017-04-18 | 2017-08-18 | 四川龙蟒福生科技有限责任公司 | The recoverying and utilizing method of accessory substance in a kind of S abscisic acids production |
| WO2018192529A1 (en) * | 2017-04-18 | 2018-10-25 | 四川龙蟒福生科技有限责任公司 | Method for preparing s-abscisic acid |
| CN116730831A (en) * | 2023-07-03 | 2023-09-12 | 江西新瑞丰生化股份有限公司 | Extraction method of abscisic acid mother liquor |
| CN116730831B (en) * | 2023-07-03 | 2024-04-12 | 江西新瑞丰生化股份有限公司 | Extraction method of abscisic acid mother liquor |
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Application publication date: 20160330 |