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SYNTHESIS OF ETHYL 3-HYDROXYBUTYRATE: A BRIEF STUDY ON THE STEREOSELECTIVE REDUCTION OF ETHYL ACETOACETATE
INTRODUCTION
This paper will conduct a brief discussion about the reduction of ethyl acetoacetate to ethyl 3-hydroxybutyrate. We have designed three individual experiments in which the reduction of ethyl acetoacetate is carried out using different reducing agents. In both experiment A and B, the reaction is carried out using sodium borohydride, while (L)-tartaric acid is introduced in B to produce a chiral environment. In experiment C, dried yeast is applied as a reductase for the starting molecule. Based on the experimental results we are looking into the effect of the reducing agent on the reaction and the product as well.
RESULTS AND DISCUSSIONS
In section A, the yield was unexpectedly low with a prolonged boil range (238~257℃ at atmospheric pressure). The major cause might have been the indiscriminate use of ethanol as the solvent. Removal of ethanol is required because it is impossible to extract the product from the aqueous layer by dichloromethane without eliminating this water-miscible organic solvent. In such step, although the temperature of the water bath was strictly controlled not exceeding 50℃, further dehydration of the product to give α,β-unsaturated ester cannot be prevented. Furthermore, as the evaporation of ethanol proceeds, the acidity of the solution becomes too strong to retain the ester group. In this sense, a variety of by-products, such as 3-hydroxybutyric acid, ethyl 2-butenoate, and 2-butenoic acid might have been present, while very little ethyl 3-hydroxybutyrate was preserved. Some water insoluble solvent, such as THF, dichloromethane, might be much promising substitute for ethanol, in that the evaporation before extraction can be avoided. Besides, since no chiral agent had been applied, the product should bear no optical rotation.
In section B, the yield was better than that in A with a satisfactorily constant boiling point under reduced pressure. The increased yield and purity should be attributed to the use of THF instead of ethanol as the solvent. The sample resembled an optical rotation value which indicates that 58.3% R enantiomer and 41.7% S enantiomer was present, given the specific rotation of each pure enantiomer. In the first stage of the reduction in experiment B, sodium borohydride first react with (L)-tartaric acid to form an intermediate which carries both chirality and the hydride group with the evolution of hydrogen gas. This intermediate then served to reduce ethyl acetoacetate preferentially from one side of the ketone group, which gave the above results.
In section C, the yield was the best among all three experiments. The boil range was 215~221℃ (converted to atmospheric pressure). Since an enzyme was applied to be the reducing agent, the reduction had to be carried out only if the orientation of the ketone group fits into the enzyme’s active site. In this sense, the stereoselectivity should be much better than that in section B. Optical rotation value indicates that the sample has a constitution of 83.1% S enantiomer to 16.9% R enantiomer, which corroborates the above reasoning. The IR spectrum has a strong absorption at 1735.8cm-1 and a wide absorption at 3446.1cm-1 with medium intensity which implies the presence of ketone and hydroxyl group. The NMR spectra: 4.19, m, 3H, 3.4, 1H, 2.47, m, 2H, 1.28, t, 3H, 1.23, d, 3H.
EXPERIMENTALS
Reduction of ethyl acetoacetate via non-chiral reagent: Experiment A
To a 100ml three-necked bottomed flask, containing a magnetic stirrer bar, add 1.5g (40mmol) sodium borohydride and 20ml ethanol, then cool the suspension to 0℃ using an ice bath. Add a solution of 5.0g (38mmol) ethyl acetoacetate in 12ml ethanol dropwise whilst stirring. The temperature should be maintained below 5℃ during the addition. Stir the solution under ice bath for an additional 15 minutes after the addition has been completed and stir the solution for a further 15 minutes at room temperature thereafter. Then cool the solution to 0℃ and neutralize the solution to weakly acidic by 1M hydrochloric acid (no suspension should be present). Stir the mixture for a further 10 minutes. Remove the ethanol on a rotary evaporator at 50℃ or below and dissolve the remaining product in 15ml dichloromethane. Remove the organic layer and extract the aqueous layer with 2×20ml dichloromethane. Dry the combined organic layer over anhydrous magnesium sulphate. Remove the solvent under reduced pressure to produce an oil, and 0.47g transparent liquid (A) is obtained by vacuum distillation at 82~95℃, 7mmHg. The overall yield is 9.3%.
Reduction of ethyl acetoacetate under chiral environment: Experiment B
To a 100ml three-necked round bottomed flask, containing a magnetic stirrer bar, add 50ml THF, 1.5g (40mmol) sodium borohydride and whilst stirring, 6.0g (40mmol) (L)-tartaric acid. Stir for 15 minutes and then cool the suspension to 0℃ using an ice bath. Add 1.32g (10.2mmol) ethyl acetoacetate dropwise and leave the solution to stir for an hour at room temperature. Then cool the reaction to 0℃ using an ice bath and neutralize the solution to weakly acidic with no presence of suspension. Stir the mixture at room temperature for 10 minutes. Remove the organic layer and extract the aqueous layer with 2×20ml ethyl acetate. Wash the combined organic layer with 40ml saturated sodium hydrocarbonate solution, 40ml brine, then dry over anhydrous magnesium sulphate. Remove the solvent under reduced pressure to produce an oil, and 0.32g transparent liquid (B) is obtained by vacuum distillation at 52℃ (the pressure unidentifiable because of technical problem). The overall yield is 23.8%, [α]D20= -7.63° (c=0.0278 , CHCl3).
Biocatalyzed reduction of ethyl acetoacetate: Experiment C
To a 500ml round bottomed flask add 1.95g (15mmol) ethyl acetoacetate, 250ml petroleum ether (60~90), 18ml distilled water and 22.5g dried yeast. Fit a mechanical stirrer and stir the reaction vigorously for 12 hours at room temperature. Decant the liquid from the yeast to obtain the organic layer and wash the yeast with 3×30ml ethyl acetate. Dry the combined organic layer over anhydrous magnesium sulphate. Remove the solvent under reduced pressure to produce an oil, and 1.05g transparent liquid (C) is obtained by vacuum distillation at 58~62℃, 5mmHg. The overall yield is 53.0%, [α]D20= +30.45° (c= , CHCl3), with IR spectra obtained.
REFERENCE
1. Dyson Perrins Lab, University of Oxford 2nd year organic practical course.
2. Lange's handbook of chemistry(15th ed.). John A. Dean. 1999.
Sample A was combined, and distilled together with Jinjing, Liu; sample C with Peng, Zhao.
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