Preparation method of 11alpha-hydroxyl-18-normethisterone
A methylnandrolone, hydroxyl technology, applied in the field of microbiology, can solve the problems of complicated purification, low volume/time efficiency, and many types of by-products, and achieves a simple separation and purification method, excellent mass transfer effect, and excellent dissolution rate. Effect
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[0032] The preparation method of 11α-hydroxy-18-methylnandrolone in one embodiment includes the following steps S110-S130:
[0033] S110, pretreating 18-methylnandrolone.
[0034] Specifically, crush 18-methylnandrolone to 100-200 meshes, add defoamer and water, and inactivate it by incubating at 60°C-90°C for 1-3 hours.
[0035] Wherein, the defoamer is a polyether type defoamer. Preferably, the defoamer is polyphenylene ether (PPE).
[0036] Further, the dosage of 18-methylnandrolone relative to the Metarhizium anisopliae is 5g / L-20g / L, and the dosage of the antifoaming agent relative to the Metarhizium anisopliae is 0.1g / L-0.5g / L.
[0037] Further, the amount (volume) of water added is 4-8 times the mass of 18-methylnandrolone.
[0038] Crush 18-methylnandrolone and mix it with defoamer and water, heat it at 60°C-90°C for 1-3 hours, not only can kill more than 99% of the miscellaneous bacteria, reduce the risk of bacterial contamination, but also can Promote its dissolu...
Embodiment 1
[0079] Embodiment 1: strain screening
[0080] 1. Test bacteria
[0081] Rhizopus nigericans, Aspergillus ochraceus, Metarhizium anisopliae
[0082] 2. Medium
[0083] Metarhizium anisopliae
[0084] Slant medium: potato cubes 300g / L, glucose 20g / L, agar 15g / L, pH natural.
[0085] Liquid medium: corn steep liquor 10g / L-20g / L, glucose 10g / L-40g / L, pH=5.5.
[0086] Rhizopus niger
[0087] Slant medium: potato cubes 300g / L, glucose 20g / L, agar 15g / L, pH natural.
[0088] Liquid medium: corn steep liquor 10g / L-20g / L, glucose 10g / L-40g / L, pH=5.5.
[0089] Ochra
[0090] Slant medium: potato cubes 300g / L, glucose 20g / L, agar 15g / L, pH natural.
[0091] Liquid medium: corn steep liquor 10g / L-20g / L, glucose 10g / L-40g / L, pH=5.5.
[0092] 3. Test method
[0093] Pick a ring of various bacteria and inoculate them on the slant medium, culture at 26°C-29°C for 6-7 days, after the spores grow, wash the spores with sterile water to make 2-9×10 8 The spore suspension per ml was us...
Embodiment 2
[0101] (1) Preparation of Metarhizium anisopliae bacteria liquid:
[0102] With reference to the cultivation method of the seed liquid in Example 1, the seed liquid was obtained.
[0103] Fermentation medium: corn steep liquor 10g / L-20g / L, glucose 10g / L-40g / L, pH value 3.8-5.5.
[0104] Inoculate 600ml seed liquid into 5.4L fermentation medium, at 30°C, 150rpm, air flow 0.2Nm 3 / h, tank pressure 0.05MPa fermentation culture for 24 hours, to obtain Metarhizium anisopliae bacterium liquid.
[0105] (2) Pretreatment of 18-methylnandrolone:
[0106] Dissolve 30g of 18-methylnandrolone crushed to 100-200 meshes and 3g of PPE in water, inactivate by incubating at 60°C-90°C for 1-3 hours, and cool for later use.
[0107] (3) Microbial transformation:
[0108] The above-mentioned pretreated 18-methylnandrolone was mixed with 6L of Metarhizium anisopliae at 30°C, the stirring speed was 150rpm, and the air flow rate was 0.2Nm 3 / h at a tank pressure of 0.05MPa for 24 hours to obtai...
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