| Androstenedione is an important intermediate of steroid hormone drugs,because of its stronger hydrophobicity,currently produced through soybean oil and water two-phase fermentation system by microbial transformation of phytosterols,lower extraction yield causing a lot of androstenedione residues in waste mother liquor.This study explores a new technology of recovery of androstenedione from the industrial waste mother liquor,mainly focusing on the organic solvent extraction,saponification oil removal,composite solvent and other process conditions.Applying liquid chromatography-mass spectrometry technology,combined with biological transformation pathway,the structure of main impurities in waste mother liquor has carried on the appraisal,main conclusions are as follows:(1)The solubilities of androstenedione in different organic solvent and soybean-oil saturated organic solvents were determined.Based on the equilibrium method,the solubilities of androstenedione in six organic solvents(methanol,acetonitrile,acetone,isopropyl alcohol,ethyl acetate and ethanol)and soybean-oil-saturated organic solvents were determined,with the temperature range from 253.15 K to 313.15 K.The results show that the solubility of androstenedione in the acetonitrile is the largest,the solubility decreases in a sequence of acetonitrile,acetone,ethyl acetate,methanol,ethanol,isopropyl alcohol.While solubilities in soybean-oil-saturated organic solvents were greater than those in the pure solvent solubility,respectively.Then the solubilities’ data were correlated with empirical equation,polynomial semi-empirical correlation model and the Apelblat equation,respectively,and all the correlation coefficients are above 98%.(2)The effect of different deoiling temperature and the extraction ratio of organic solvent-oil phase on the oil remove and recovery has been studied.And the optimum deoiling conditions was acetonitrile as extraction solvent,when a temperature of 253.15 K,and extraction volume ratio 2:1,the oil removal rate was 41.14%,the per liter waste mother liquor can recycle 1.82 g androstenedione.The optimal saponification conditions was determined by orthogonal experimental design method,that was adding 10% Na OH1.5g per liter solution and reacting at 343.15 K temperatures for 80 minutes,the recycled coarse products was 1.38 g,with the purity of 56.13%.(3)The purification process of coarse products was studied.Using single factor experiment,the recovery rate and purity of product as examining index,the compound solvent type,the proportion of compound solvent in organic solvent with n-hexane and the addition amount of compound solvent were determined.Subsequently applying orthogonal experimental scheme,the optimal refined process conditions was the compound solvent mixture of ethanol-hexane in a ratio of 1:3,2 times the amount of adding volume,under the condition of 278.15 K for purification.The recovery rate of the product was 52.07% and the purity was 83.52%.Finally,organic solvent crystallization method was used to improve the purity of the product,and the recovery rate of androidone was 33.46% and the purity reached 98.58%,which met the requirements of industrial production.(4)The composition and structure of 7 kinds of substances in waste mother liquor were demonstrated by using chromatography-mass spectrometry technology.Among them,component A can be identified to androsta-1,4-diene-3,17-dione as structural analogue of androstenedione;component B is targeted product androstenedione;component C may be the degradation products of androstenedione under the action of9α-hydroxylase;group D is a mixture,including three kinds of material,which are likely to androstenedione intermediate products of biological transformation;component E is corresponding to the substance of incompletely degraded side-chain of the plant sterol. |
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