CN104231043B - The method that high-speed countercurrent chromatography purifies Nosiheptide - Google Patents
The method that high-speed countercurrent chromatography purifies Nosiheptide Download PDFInfo
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- CN104231043B CN104231043B CN201410396680.3A CN201410396680A CN104231043B CN 104231043 B CN104231043 B CN 104231043B CN 201410396680 A CN201410396680 A CN 201410396680A CN 104231043 B CN104231043 B CN 104231043B
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- nosiheptide
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- countercurrent chromatography
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Abstract
The invention discloses a kind of method of extraction purification Nosiheptide.Method is using Nosiheptide mycelium as raw material, tetrahydrofuran is used as Extraction solvent ultrasonic extraction 2 times, extract solution elder generation peroxidating aluminium short column, collected post liquid, most of solvent is fallen in concentration, and gained extraction cream is separated using high-speed countercurrent chromatography, UV-detector on-line checking, target component is collected, is dried under reduced pressure to obtain the Nosiheptide of high-purity.Nosiheptide is purified using this method, simple to operate, short preparation period, solvent is recyclable, and overall recovery can reach 80%, and the Nosiheptide content of acquisition is not less than 95%, and highest content can reach 99%, and tool has great advantage compared with conventional method.
Description
Technical field
The present invention relates to a kind of method of extraction purification Nosiheptide, is purified more particularly to a kind of high-speed countercurrent chromatography
The method of Nosiheptide.
Background technology
Nosiheptide is animal specific antibiotic, and its Antibacterial mechanism is to suppress bacterioprotein synthesis and play antibacterial and make
With.Bacterium is not likely to produce drug resistance to Nosiheptide, with other antibiotic also without obvious cross resistance.Nosiheptide is to gram sun
Property bacterium, particularly staphylococcus have higher activity.Nosiheptide small toxicity, mixed feeding administration seldom absorb in animal alimentary canal,
Thus remained less in animal product, be primarily to facilitate growth of animals or poultry.
Nosiheptide molecular formula：
Molecular formula：C51H43N13O12S6
Molecular weight：1222.37
Character：This product is light yellow green brown or yellowish green brown powder, there is characteristic odor
Existing to take wet underwater welding, then obtain product by separation of solid and liquid, it is low that technical problem essentially consists in overall recovery,
Only 50%-60%, Nosiheptide decomposes more in extraction process, obtain Nosiheptide product content it is not high can only be to 70%.It is molten
Agent consumption is bigger, and environmental pollution is serious.
The content of the invention
It is contemplated that the defects of overcoming prior art and deficiency, there is provided a kind of side of efficient extraction purification Nosiheptide
Method, obtain the Nosiheptide fine work of high-purity.
The purpose of the present invention is achieved through the following technical solutions：
The method that high-speed countercurrent chromatography purifies Nosiheptide, methods described are carried out as follows：(1) with the dry bacterium of Nosiheptide
Filament is raw material, uses tetrahydrofuran to be extracted several times using ultrasonic wave extraction mode for Extraction solvent；(2) step is merged
Suddenly (1) extract solution, by extract solution peroxidating aluminium short column, post liquid was collected；(3) the post liquid concentration of crossing of step (2) is extracted
Cream, gained extraction cream are separated using high-speed countercurrent chromatography, UV-detector on-line checking, collect target component, decompression
It is dried to obtain the Nosiheptide of high-purity.
Preferably, step (3) the high speed counter current chromatography separation application is：Take ethyl acetate, methanol, stone
Oily ether and water are sufficiently mixed according to the ratio of volume ratio 1-5: 1-7: 1-3: 1-7, stratification, take phase to fill high speed adverse current chromatogram
Pipe, main frame is rotated with 900-1100rpm, lower phase is pumped into and does mobile phase, after reaching dynamic equilibrium, flow rate of mobile phase is arranged to 1-
5ml/min, with flowing phased soln extraction cream sample introduction, composition is collected according to collection of illustrative plates.
Preferably, post processing extraction solvent consumption is 3-5 times/time of the dry mycelia weight of Nosiheptide in the step (1), extraction
Number is 2-3 times, crosses when solvent is fallen in the concentration of post liquid and concentrates 12-20 times.
Preferably, the aluminum oxide in the step (2) is neutral alumina, granularity is 200-300 mesh.
Preferably, using being concentrated in vacuo, temperature is less than 50 degrees Celsius for concentration in the step (2).
Preferably, in the step (3) target component pressure be dried to obtain Nosiheptide drying temperature be less than 50 degrees Celsius,
Vacuum is less than 1000Pa.
Beneficial effects of the present invention：The present invention is to the extraction purification efficiency high of Nosiheptide, and the rate of recovery of Nosiheptide is high, that west
The purity of peptide is high.
Embodiment
Technical scheme is described further below by specific embodiment, so reagent in embodiment
It is conventional reagent purchased in market.
Nosiheptide mycelia, Zhejiang Province Huineng animal drugs Co., Ltd's fermentation obtain.
Short column of neutral alumina：Huadong Medicine Co., Ltd..
High-speed counter-current chromatograph, Jiangyin Countercurrent Technology Co., Ltd., model OptiChrome-1000.
Embodiment 1
The method that high-speed countercurrent chromatography purifies Nosiheptide, methods described are carried out as follows：Nosiheptide mycelium dries
After dry, 1000g (Nosiheptide content 10%) is taken, adds 3 times of amount (3000ml) tetrahydrofuran solvent ultrasonic wave extractions 15 minutes, mistake
Filter, then measure (3000ml) tetrahydrofuran ultrasonic wave extraction 15 minutes with 3 times, filtering, merging filtrate, extract solution crosses neutral alumina
Short column (granularity 200-300 mesh), cross 12 times of post liquid reduced vacuum concentration and obtain extraction cream to 500ml, thickening temperature is less than or waited
In 50 degrees Celsius；
Take ethyl acetate, methanol, petroleum ether, water is sufficiently mixed according to 1: 3: 1: 3 ratios, stratification, takes phase to fill
High speed adverse current chromatogram pipe, main frame is rotated with 900rpm, lower phase is pumped into and does mobile phase, after reaching dynamic equilibrium, flow rate of mobile phase is set
3ml/min is set to, extraction cream sample introduction is dissolved according to volume ratio 10: 1 with mobile phase, composition is collected according to collection of illustrative plates；
Reclaim reagent, it is dried under reduced pressure, vacuum drying vacuum is 800Pa, 45 degrees Celsius of temperature, until solvent evaporated obtains
To Nosiheptide 83g, content 97.3%, the rate of recovery 80.75%.
Embodiment 2
The method that high-speed countercurrent chromatography purifies Nosiheptide, methods described are carried out as follows：Nosiheptide mycelium dries
After dry, 1000g (Nosiheptide content 10%) is taken, adds 5 times of amount (5000ml) tetrahydrofuran solvent ultrasonic wave extractions 15 minutes, mistake
Filter, then measure (5000ml) tetrahydrofuran ultrasonic wave extraction 15 minutes with 5 times, filtering, merging filtrate, extract solution crosses neutral alumina
Short column (granularity 200-300 mesh), cross 202 times of post liquid reduced vacuum concentration and obtain extraction cream to 500ml, thickening temperature is less than or waited
In 50 degrees Celsius.
Take ethyl acetate, methanol, petroleum ether, water is sufficiently mixed according to 1: 5: 1: 5 ratios, stratification, takes phase to fill
High speed adverse current chromatogram pipe, main frame is rotated with 1100rpm, lower phase is pumped into and does mobile phase, after reaching dynamic equilibrium, flow rate of mobile phase is set
5ml/min is set to, extraction cream sample introduction is dissolved according to volume ratio 10: 1 with mobile phase, composition is collected according to collection of illustrative plates.
Reclaim reagent, it is dried under reduced pressure, vacuum drying vacuum is 800Pa, 45 degrees Celsius of temperature, until solvent evaporated,
Obtain Nosiheptide 88.3g, content 98.1%, the rate of recovery 86.62%.
Embodiment 3
The method that high-speed countercurrent chromatography purifies Nosiheptide, methods described are carried out as follows：Nosiheptide mycelium dries
After dry, 1000g (Nosiheptide content 10%) is taken, adds 5 times of amount (5000ml) tetrahydrofuran solvent ultrasonic wave extractions 30 minutes, mistake
Filter, then measure (3000ml) tetrahydrofuran ultrasonic wave extraction 30 minutes with 3 times, filtering, merging filtrate, extract solution crosses neutral alumina
Short column (granularity 200-300 mesh), cross 16 times of post liquid reduced vacuum concentration and arrive 500ml, thickening temperature is less than or equal to 50 degrees Celsius.
Take ethyl acetate, methanol, petroleum ether, water is sufficiently mixed according to 1: 3: 1: 3 ratios, stratification, takes phase to fill
High speed adverse current chromatogram pipe, main frame is rotated with 1000rpm, lower phase is pumped into and does mobile phase, after reaching dynamic equilibrium, flow rate of mobile phase is set
5ml/min is set to, extraction extraction cream sample introduction is dissolved according to volume ratio 10: 1 with mobile phase, composition is collected according to collection of illustrative plates.
Reclaim reagent, it is dried under reduced pressure, vacuum drying vacuum is 800Pa, 45 degrees Celsius of temperature, until solvent evaporated,
Obtain Nosiheptide 85g, content 95.3%, the rate of recovery 81%.
Embodiment described above is a kind of preferable scheme of the present invention, not the present invention is made any formal
Limitation, there are other variants and remodeling on the premise of without departing from the technical scheme described in claim.
Claims (1)
1. the method that high-speed countercurrent chromatography purifies Nosiheptide, it is characterised in that：Methods described is carried out as follows：(1) with
The dry mycelium of Nosiheptide is raw material, uses tetrahydrofuran to be extracted several times using ultrasonic wave extraction mode for Extraction solvent；
(2) combining step (1) extract solution, by extract solution peroxidating aluminium short column, post liquid was collected；(3) step (2) is crossed into the concentration of post liquid
Obtain extraction cream, gained extraction cream separated using high-speed countercurrent chromatography, UV-detector on-line checking, collect target into
Point, it is dried under reduced pressure to obtain the Nosiheptide of high-purity；
Extraction solvent volumetric usage is 3-5 times/time of the dry mycelia weight of Nosiheptide in the step (1), extraction time 2-3
It is secondary, cross when solvent is fallen in the concentration of post liquid and concentrate 12-20 times；
Aluminum oxide in the step (2) is neutral alumina, and granularity is 200-300 mesh, and using being concentrated in vacuo, temperature is small for concentration
In 50 degrees Celsius；
Step (3) the high speed counter current chromatography separation application is：Take ethyl acetate, methanol, petroleum ether and water according to
Volume ratio 1-5:1-7:1-3:1-7 ratios are sufficiently mixed, stratification, take phase to fill high speed adverse current chromatogram pipe, with 900-
1100rpm rotates main frame, is pumped into lower phase and does mobile phase, and after reaching dynamic equilibrium, flow rate of mobile phase is arranged to 1-5ml/min, uses
Phased soln extraction cream sample introduction is flowed, composition is collected according to collection of illustrative plates；
In the step (3) target component pressure be dried to obtain Nosiheptide drying temperature be less than 50 degrees Celsius, vacuum is less than
1000Pa。
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105420319A (en) * | 2015-12-24 | 2016-03-23 | 山东胜利生物工程有限公司 | Preparation method of pure nosiheptide |
| CN106317174A (en) * | 2016-08-25 | 2017-01-11 | 宁夏泰益欣生物科技有限公司 | Preparing method of crude nosipeptide |
| CN106632593A (en) * | 2016-12-22 | 2017-05-10 | 甘肃汇能生物工程有限公司 | Extraction method of nosiheptide fine powder |
| CN113388001B (en) * | 2020-03-11 | 2023-04-07 | 深圳翰宇药业股份有限公司 | Method for purifying octreotide acetate by high-speed counter-current chromatography |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4384043A (en) * | 1981-09-02 | 1983-05-17 | American Cyanamid Company | Process for producing the antibiotic nosiheptide |
| CN1840684A (en) * | 2006-01-26 | 2006-10-04 | 浙江工商大学 | Process for fermentation preparation of nosiheptide by using streptomycete |
-
2014
- 2014-08-13 CN CN201410396680.3A patent/CN104231043B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4384043A (en) * | 1981-09-02 | 1983-05-17 | American Cyanamid Company | Process for producing the antibiotic nosiheptide |
| CN1840684A (en) * | 2006-01-26 | 2006-10-04 | 浙江工商大学 | Process for fermentation preparation of nosiheptide by using streptomycete |
Non-Patent Citations (1)
| Title |
|---|
| 高速逆流色谱研究进展;戴德舜等;《分析化学评述与进展》;20010525;第29卷(第5期);586-591 * |
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Address after: Changan city Jiaxing town Zhejiang city of Haining province (314400 agricultural areas) Chunchao Road No. 3 Applicant after: ZHEJIANG ESIGMA BIOTECHNOLOGY CO., LTD. Address before: Changan city Jiaxing town Zhejiang city of Haining province (314400 agricultural areas) Chunchao Road No. 3 Applicant before: Zhejiang Huineng Animal Medicine Co., Ltd. |
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