Chemical Modification And Biological Transformation Of Plant Steroid

19-hydroxy androstendione(C19-OH-AD)is a kind of important pharmaceutical intermediate,which is mainly synthesized by chemical synthesis method.This process is complex,polluted and the yield is low.So,the synthesis of 19--hydroxy-androstendione and its related technology were studied by using the methods of chemical synthesis and microbial transformation.First,plant sterol materials synthesis 6,19-epoxy acetyl intermediates after three step reactions:acetyl,addition and oxidation,then we can obtain 6,19-epoxy androstenedione by microbial transformation,6,19-epoxy androstenedione synthesis 19 hydroxyl androstene Dione after ring opening reaction.Specific research results are as follows:Acetylation:plant sterols as raw material,toluene as solvent,acetic anhydride as acetylation reagent.Suitable synthesis conditions are: reaction temperature is 90 ?,reaction time 3h,raw material ratio is m(sterols): m(acetic anhydride)= 1:2.16 and solvents added amount is 9 times(quality)of phytosterols,the yield can reach 92.5% by TLC analysis.Addition reaction:acetone as the solvent,acetylation product as raw material,calcium hypochlorite and acetic acid as additive agent.Suitable synthesis conditions: reaction temperature is 20 ?,reaction time 45 minute,feeding mode is added in batches,the ratio of raw materials is M(acetyl sterols):m(calcium hypochlorite):m(30% acetic acid)=1:0.5:1.21,solvent amount is 24 times(quality)of acetyl sterol.The yield was 60.2% by TLC analysis.Cyclic oxidation reaction: addition product as the raw material,cyclohexane as the solvent,the chloride butyl two as ring oxidation reagent.Suitable process conditions are:reaction temperature is 80 ?,reaction time 4.5 hours,400 W light reaction,raw material ratio is m(chlorine alcoholate): m(chlorinated butyl acyl imine)= 1:0.5,amount of solvent is 27times(quality)of chlorine alcoholization.The yield was 85.12% by TLC analysis.The high performance liquid chromatography(HPLC)operating conditions of Microbialtransformation: C18 alkyl silane bonded reversed-phase column.The mobile phase is V(methanol): V(acetonitrile): V(water)=35:35:30;sample size is 20 ? L;volume flow is0.8ml / min;column temperature is room temperature;UV detector wavelength is 242 nm.The appropriate process conditions for microbial transformation: the amount of bottled liquid is 50ml/500 ml.Substrate addition amount is 2g/L,inoculation amount is 8%,fermentation temperature 30 degrees Celsius,fermentation time 72 h,the yield can reach more than 95%.Opening ring reaction: microbial transformation products as raw materials,ethanol as solvent,zinc and acetic acid as open-loop reagent,optimum conditions of the reaction:reaction temperature is 80 ?,reaction time is 45 min,the ratio of raw materials is M(6,19 of epoxy androstene two ketone): m(acetic acid): m(Zn)=1: 0.63:1.21 and solvents added amount is 12 times(quality)of microbial conversion product,by liquid chromatography analysis,the yield can reach as high as 60%.