CN102584772A - Method for preparing ochratoxin B - Google Patents
Method for preparing ochratoxin B Download PDFInfo
- Publication number
- CN102584772A CN102584772A CN2011100008048A CN201110000804A CN102584772A CN 102584772 A CN102584772 A CN 102584772A CN 2011100008048 A CN2011100008048 A CN 2011100008048A CN 201110000804 A CN201110000804 A CN 201110000804A CN 102584772 A CN102584772 A CN 102584772A
- Authority
- CN
- China
- Prior art keywords
- ochratoxin
- methanol
- extract
- stream part
- stream
- 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
Images
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a method for preparing ochratoxin B, wherein the ochratoxin B is obtained by separating and purifying the rice culture of ochratoxin high-yield strains through applying antiphase medium-pressure preparation chromatography, gel LH-20 column chromatography, antiphase high-pressure preparation chromatography and other technologies. The technology is simple, the yield is improved, the cost is reduced, and the industrial production is greatly benefited.
Description
Technical field:
The present invention relates to the method for separation and purification ochratoxin B (OTB) from the rice culture of ochratoxin superior strain.
Background technology:
Ochratoxin (Ochratoxins) be one type to the healthy deleterious fungus metabolite of humans and animals; Mainly under the adapt circumstance condition, produce by Aspergillus ochraceus (Aspergillus) and mould (Penicillium); Substruction is that Isocoumarin >97 is connected to the verivate on β-phenylalanine(Phe), and 7 types of A, B, C, D etc. are arranged.
At present; The problem of ochratoxin contaminated food products and Chinese medicinal materials is on the rise; In order to guarantee the supply of the required ochratoxin standard substance of check and analysis, obtain a large amount of, highly purified ochratoxin, we have selected the Aspergillus ochraceus of high yield to carry out the rice cultivation; Therefrom separation of pure dissolves highly purified ochratoxin B, for the further investigation of this mycotoxins provides the foundation.
Summary of the invention:
The method that the purpose of this invention is to provide separation and purification ochratoxin B from the rice culture of ochratoxin superior strain.
Technical scheme of the present invention comprises following method steps successively:
1. screening superior strain
2. separation and purification ochratoxin B
Embodiment:
The following embodiment of institute helps those skilled in the art to understand the present invention better, but does not limit the present invention in any way.
1. screening superior strain
Working foundation according to our early stage; Investigate the influence of culture condition (temperature, humidity, illumination, pH value, carbon source, nitrogenous source etc.), optimize culture condition, the superior strain of screening product ochratoxin toxigenic bacterium strain growth and product poison amount; Cultivate through rice, obtain culture.
2. separation and purification ochratoxin B
Suppress technology such as being equipped with chromatogram, gel LH-20 column chromatography and anti-phase high pressure preparative hplc in the application anti-phase ochratoxin in this culture is separated and purifying, set up the preparation method of ochratoxin B.
Rice culture (2kg) to the ochratoxin superior strain is pulverized, the ethanolic soln heating and refluxing extraction with 80% 4 times, and each extraction time is 2 hours; United extraction liquid is evaporated to when not having the alcohol flavor; Change in the separating funnel, adopt equal-volume ETHYLE ACETATE to extract, extract four times after; Combining extraction liquid, concentrating under reduced pressure obtains acetic acid ethyl ester extract.This extract is dissolved in the methyl alcohol, removes by filter insolubles, after filtrating concentrates, adopt in the anti-phase compacting to be equipped with chromatogram and separate; With 30%, 50%, 70% and 100% methanol/water solution gradient elution, each stream part of thin layer inspection merges stream part of containing blue-fluorescence, behind the stream part thickening filtration after the merging successively; Adopt gel LH-20 column chromatography to separate, methanol-eluted fractions obtains containing the bullion of toxin; This bullion is carried out reversed phase high efficiency liquid phase preparative hplc separate, each stream part of thin layer inspection is reclaimed solvent; Obtain white solid, after recrystallizing methanol, obtain clear crystal 2mg.
Ochratoxin B (Ochratoxin B)
[1], molecular formula is C
20H
19NO
6, molecular weight is 369.ESI-MS：m/z?370[M+H]
+，392[M+Na]
+，761[2M+Na]
+；
1H?NMR(CDCl
3，600MHz)δ：8.32(1H，d，J＝7.8Hz，H-7)，7.21-7.29(5H，m，H-17～21)，6.83(1H，d，J＝8.4Hz，H-6)，4.99(1H，m，H-3)，4.76(1H，td，J＝6.0，4.2Hz，H-14)，3.32(1H，dd，J＝14.4，5.4Hz，H-15b)，3.19(1H，J＝13.8，6.6Hz，H-15a)，2.99(1H，m，H-4b)，2.97(1H，m，H-4a)，1.55(3H，d，J＝6.0Hz，H-11)；
13C?NMR(CDCl
3，150MHz)δ：173.6(C-22)，170.3(C-1)，164.2(C-12)，160.4(C-9)，143.9(C-5)，138.9(C-16)，136.4(C-7)，129.5(C-17，21)，128.6(C-18，20)，127.1(C-19)，119.2(C-6)，118.7(C-8)，108.8(C-10)，76.4(C-3)，54.3(C-14)，37.7(C-15)，34.7(C-4)，20.7(C-11)。
Instrument and reagent
Bruker AV-600 type NMR spectrometer with superconducting magnet; LTQ Orbitrap XL mass spectrograph; Germany LUMTECH liquid chromatograph K-1001; Thin layer plate GF
254(factory of Haiyang Chemical Plant, Qingdao article); Reagent is analytical pure (Beijing Chemical Plant).
The thin layer condition
Developping agent: 10: 1: 0.05 (v: v: v) of chloroform/methanol/formic acid; Detect wavelength 254nm and 365nm.
Chromatographic condition
Chromatographic column: Kromasil MZ-C
18(10 μ m, 250 * 10mm); Moving phase: methanol, gradient elution; Flow velocity: 2ml/min; Column temperature: normal temperature; Detect wavelength: 333nm.
Reference
[1]M.W.Bredenkamp，J.L.M.Dillen，P.H.van?Rooyen，P.S.Steyn.Crystal?Structures?and?Conformational?Analysis?of?Ochratoxin?A?and?B：Probing?the?Chemical?Structure?causing?Toxicity.J.CHEM.SOC.PERKIN?TRANS.II，1989，1835-1839.
Description of drawings
The structure of Fig. 1-ochratoxin B
Fig. 2-separation process
The hydrogen spectrum of Fig. 3-ochratoxin B
The carbon spectrum of Fig. 4-ochratoxin B
Claims (1)
1. the preparation method of an ochratoxin B, it is characterized in that: the rice culture to the ochratoxin superior strain is pulverized, and with 80% ethanolic soln heating and refluxing extraction 4 times, each extraction time is 2 hours; United extraction liquid is evaporated to does not have the alcohol flavor, changes in the separating funnel, and equal-volume ETHYLE ACETATE extracts, extract four times after; Combining extraction liquid, concentrating under reduced pressure obtains acetic acid ethyl ester extract, and this extract is dissolved in the methyl alcohol, removes by filter insolubles; After filtrating concentrates, adopt in the anti-phase compacting to be equipped with chromatogram and separate, successively with 30%, 50%, 70% and 100% methanol/water solution gradient elution, each stream of thin layer inspection part; Stream part to containing blue-fluorescence merges, and behind the stream part thickening filtration after the merging, adopts gel chromatography separation, methanol-eluted fractions; Obtain containing the bullion of toxin, this bullion is carried out anti-phase high pressure preparative hplc separate, each stream part of thin layer inspection merges the back and reclaims solvent; Obtain white solid, after recrystallizing methanol, obtain ochratoxin B (OTB).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100008048A CN102584772A (en) | 2011-01-05 | 2011-01-05 | Method for preparing ochratoxin B |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011100008048A CN102584772A (en) | 2011-01-05 | 2011-01-05 | Method for preparing ochratoxin B |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102584772A true CN102584772A (en) | 2012-07-18 |
Family
ID=46474097
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011100008048A Pending CN102584772A (en) | 2011-01-05 | 2011-01-05 | Method for preparing ochratoxin B |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102584772A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604674A (en) * | 2013-11-21 | 2014-02-26 | 四川理工学院 | Method for extracting and purifying ochratoxin A from yeast for making hard liquor |
| CN104178535A (en) * | 2014-04-09 | 2014-12-03 | 上海市农业科学院 | Method for preparing and purifying ochratoxin A and ochratoxin B |
| CN115491316A (en) * | 2022-07-01 | 2022-12-20 | 上海市农业科学院 | Preparation method of concealed toxin T-2-3G |
-
2011
- 2011-01-05 CN CN2011100008048A patent/CN102584772A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103604674A (en) * | 2013-11-21 | 2014-02-26 | 四川理工学院 | Method for extracting and purifying ochratoxin A from yeast for making hard liquor |
| CN104178535A (en) * | 2014-04-09 | 2014-12-03 | 上海市农业科学院 | Method for preparing and purifying ochratoxin A and ochratoxin B |
| CN115491316A (en) * | 2022-07-01 | 2022-12-20 | 上海市农业科学院 | Preparation method of concealed toxin T-2-3G |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhu et al. | Penochalasin K, a new unusual chaetoglobosin from the mangrove endophytic fungus Penicillium chrysogenum V11 and its effective semi-synthesis | |
| CN109705188B (en) | Triterpenoid compound in exocarpium Juglandis Immaturum, and preparation method and application thereof | |
| CN106496171B (en) | Polycyclic polyisocyanate pentenyl phloroglucinol derivatives compound, preparation method and application with anti-tumor activity | |
| CN106589020B (en) | A method of extracting icariin from Herba Epimedii | |
| CN102584772A (en) | Method for preparing ochratoxin B | |
| CN103088083A (en) | Method for preparing anti-inflammation and anti-tumor activity sesquiterpene lactone compounds | |
| CN106243078B (en) | The preparation method and purposes of compound in Fu Kangshu | |
| CN105061545B (en) | Triterpene saponin componds and its preparation method and application in shiny-leaved yellowhorn | |
| CN110092809B (en) | Method for separating and extracting beta-sitosterol by using bacillus megaterium | |
| Ning et al. | Cytotoxic biotransformed products from triptonide by Aspergillus niger | |
| CN104140391B (en) | A kind of method of preparing lathyrol oxalic acid nicotinate that separates from moleplant seed | |
| Ma et al. | A New Peroxy‐multiflorane Triterpene Ester from the Processed Seeds of Trichosanthes kirilowii | |
| CN114621224B (en) | Maca alkaloid and preparation method and application thereof | |
| Yan et al. | New serratene triterpenoids from Palhinhaea cernua and their cytotoxic activity | |
| CN102532243B (en) | Method for simultaneously preparing multiflora rose glycoside and rose glycoside compounds | |
| CN101798294B (en) | Preparation method of anti-tumour medicine intermediate 10-deacetylbacctin III | |
| CN105420336B (en) | A method of Vanguerolic acid-A is converted by rotundic acid using microorganism | |
| Zhou et al. | Two compounds from the endophytic Colletotrichum sp. of Ginkgo biloba | |
| CN104004042B (en) | A kind of steroid compound and preparation method thereof and the application as antivirotic | |
| CN106565661B (en) | The method for isolating and purifying nujiangexanthoneA | |
| Liu et al. | Preparation of progenin III from total steroidal saponins of Dioscorea nipponica Makino using a crude enzyme from Aspergillus oryzae strain | |
| CN102321060A (en) | Method for extracting scopoletin from artemisia annua residues | |
| CN113024498A (en) | Ficus-japonica alkane sesterterpene compound, preparation method thereof, pharmaceutical composition and application thereof, and application of total extract of Ficus-japonica | |
| CN110938105A (en) | Extraction and separation method of active ingredients of agaricus tabularis | |
| CN102584849A (en) | Method for preparing aflatoxin B1 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | 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: 20120718 |