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Method for separating purified 11 Alpha-hydroxy canrenone

A technology for the separation and purification of hydroxycanrenone, which is applied in the fields of organic chemistry and steroids, can solve the problems of cumbersome purification steps of 11α-hydroxycanrenone, and achieve the effect of high purity, simple procedure steps and simple purification steps

Inactive Publication Date: 2014-03-26
JIANGSU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This requires a large amount of toxic organic solvents, and the purification steps of 11α-hydroxycanrenone after extraction are cumbersome

Method used

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  • Method for separating purified 11 Alpha-hydroxy canrenone

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Wash the rhizopus slant covered with spores for 7 days with 15 mL of sterile water to obtain a spore suspension. Add 1mL of spore liquid into a 500mL Erlenmeyer flask containing 50mL of fresh sterile medium. After cultivating at 25°C and 220r / min for 48h, add canrenone so that the concentration of canrenone in the fermentation broth is 1g / L, continue After 48 hours of cultivation and transformation, the transformation rate was measured to be 91.3%, and the transformation was stopped. Among them: Rhizopus slant medium components include (g / L): glucose 22.0, yeast extract 20.0, soybean peptone 20.0, agar 30.0, adjust the pH of the medium to 5.0 with 10% phosphoric acid solution; liquid transformation medium group Include (g / L): glucose 22.0, yeast extract 16.5, corn steep liquor 33.0.

[0020] (2) Centrifuge the converted fermentation broth at 1000rpm for 30 minutes, dry the mycelium at 40°C, and crush it to 200 mesh.

[0021] (3) Extraction. Soak the pulverized my...

Embodiment 2

[0027] (1) Wash the slanted surface of Aspergillus ochrae that has been cultured for 5 days and is covered with spores with 5 mL of sterile water to obtain a spore suspension. Add 3mL of spore liquid into a 500mL Erlenmeyer flask containing 150mL of fresh sterile medium. After cultivating at 30°C and 150 r / min for 24 hours, add canrenone so that the concentration of canrenone in the fermentation broth is 5g / L. After continuing to cultivate and transform for 96 hours, the transformation rate was measured to be 96.2%, and then the transformation was stopped. Among them: Ochra slant medium components include (g / L): glucose 20.0, yeast extract 20.0, soybean peptone 20.0, agar 20.0, adjust the pH of the medium to 6.0 with a 10% phosphoric acid solution; liquid transformation medium group Include (g / L): glucose 20.0, yeast extract 25.0, corn steep liquor 2.0.

[0028] (2) After the transformed fermentation broth is vacuum filtered, the mycelium is dried at 100°C and crushed to 50 m...

Embodiment 3

[0035] (1) Wash the rhizopus slant covered with spores for 5 days with 10 mL of sterile water to obtain a spore suspension. Add 2mL of spore liquid into a 500mL Erlenmeyer flask containing 100mL of fresh sterile medium. After cultivating at 28°C and 200 r / min for 36 hours, add canrenone so that the concentration of canrenone in the fermentation broth is 5g / L. After continuing to cultivate and transform for 84 hours, the transformation rate was measured to be 93.2%, and then the transformation was stopped. Among them: Rhizopus slant medium components include (g / L): glucose 22.0, yeast extract 20.0, soybean peptone 20.0, agar 30.0, adjust the pH of the medium to 5.0 with a 10% phosphoric acid solution; liquid transformation medium group Include (g / L): glucose 22.0, yeast extract 16.5, corn steep liquor 33.0.

[0036] (2) Centrifuge the converted fermentation broth at 6000rpm for 10 minutes, take the mycelium, dry it at 80°C, and crush it to 100 mesh.

[0037] (3) Extraction. ...

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Abstract

The invention relates to a method adopting macroporous resin to separate purified 11 Alpha-hydroxy canrenone. The method comprises the following steps: mycelium separation, drying for crushing, heating methanol for extraction, macroporous resin column adsorption separation and purification, pressurizing for continuous chromatography and concentrating drying. The method has the advantages of simple technological process, low cost and high benefits, chromatographic columns can be activated and regenerated, and the method is suitable for industrially separating and purifying 11 Alpha-hydroxy canrenone. A methanol solution is used for heating extraction, so that the extraction ratio of 11 Alpha-hydroxy canrenone is higher. AB-8 macroporous resin is adopted for continuous chromatography under the driving of a high-pressure constant flow pump, and finally purer 11 Alpha-hydroxy canrenone is obtained.

Description

Technical field [0001] The patent of the present invention involves the use of large-hole resin to separate 11α-hydroxylidone, which belongs to the field of extraction and purification of microbial transformation products. Background technique [0002] 11α-hydroxylcanlidone is a bodied derivative formed after the reaction of Cumitone hydroxylated.Eprite has a broad market prospect as a new drug for treating hypertension and other cardiovascular diseases.The method of microorganisms to convert Cantidone synthesis 11α-hydroxylidone has the advantages of strong specificity, mild response conditions, high catalytic efficiency, less by-products, low cost, and less pollution. [0003] Due to the extremely low solubility of 11α-hydroxylkone in water, 11α-hydroxylkone, which is synthesized by microbial fermentation, is mainly adsorbed on the surface of the mycelium. It is difficult to separate the physical methods of filtering, centrifugation or extraction.At the same time, there are sti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07J21/00
Inventor 管国强黄达明孙文敬崔凤杰崔鹏景钱静亚
Owner JIANGSU UNIV
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