| Excessive reactive oxygen species(ROS)in the human body could damage bioactive molecules such as proteins,lipids and DNA,accelerate the body's aging,and induce multiple diseases.Some studies have shown that pyroligneous acid contains a variety of bioactive components,which had obvious antioxidant activity that could protect the body from the damage caused by oxidative stress.According to the difference of chemical polarity and existing form of pyroligneous acid,the phenolic fractions of jujube shell pyroligneous acid were separated by emulsion liquid membrane,the antioxidant and hypoglycemic activities of pyroligneous acid were investigated by the method of in vitro enzymatic activity in this thesis.The results are as follows:(1)The preparation technology of emulsion liquid membrane was studied from the following two aspects: the stability of emulsion liquid membrane and the effect of removal phenol from pyroligneous acid.The effects of emulsification time,emulsification speed,the type and volume fraction of surfactant,the volume fraction of the carrier,the concentration of internal phase Na OH and the ratio of oil phase volume to internal phase volume on the stability of the emulsion were investigated.And the effects of the volume fraction of the carrier,the concentration of internal phase Na OH and the ratio of oil phase volume to internal phase volume on the removal of phenol were investigated.Colligating the stability of emulsion and the effect of phenol removal,the experiment conditions of various factors were determined.And the volume fraction of surfactant T154,the volume fraction of the carrier,the concentration of internal phase NaOH and the ratio of oil phase volume to internal phase volume were selected as four main factors to optimize by using BBD Response Surface Method.The results show that the optimum emulsification time might be 20 min,the emulsification speed might be 4500 r/min,and the stability of the emulsion prepared by the surfactant T154 might be better than HT154 and Span 80 under the same conditions.According to variance analysis,the effects of various factors on the rate of phenol removal in pyroligneous acid were D(the ratio of oil phase volume to internal phase volume)>C(the concentration of internal phase NaOH)>B(the volume fraction of TBP)>A(the volume fraction of T154)and factors of A and C,B and C,B and D had the main interaction.The optimum preparation conditions of emulsion liquid membrane might be that the concentration of surfactant T154 is 8%,the liquid carrier TBP is 3%,the concentration of internal phase Na OH is 0.6 mol/L,the ratio of oil phase volume to internal phase volume is 5:4.Under this condition,the stability of the emulsion could be better and the rate of phenol removal from pyroligneous acid could be 55.31%.(2)During refining of pyroligneous acid by emulsion liquid membrane,the effects of extraction time,stirring speed and the ratio of emulsion to water on phenol removal were mainly investigated,and the parameters were optimized by using BBD Response Surface Method.According to variance analysis,the effects of various factors on the rate of phenol removal in pyroligneous acid were C(the ratio of emulsion to water)>A(extraction times)>B(tirring speed),and factors of A and C,A and B had the main interaction.The optimum conditions of phenol removal could be that the extraction time is 25 min,the extra ction stirring speed is 300 r/min and the ratio of emulsion to water is 1:2.Under this condition,the rate of phenol removal from pyroligneous acid could be 59.41%.(3)The methods of DPPH,ABTS,FRAP,·OH,T-AOC and total reducing capacity were used to determine the antioxidant activity of pyroligneous acid.The results showed that pyroligneous acid had good capacity of DPPH,ABTS and ·OH scavenging,strong iron ion reduction/antioxidant activity,and has strong capacity of T-AOC and total reducing.With Vc equivalent representation,the capacity of DPPH free radical scavenging of the pyroligneous acid was 0.6178±0.0107 mg Vc/m L,the capacity of ABTS free radical scavenging was 1.7547±0.0136 mg Vc/mL,the value of FRAP was 0.8100±0.0132 mg Vc/mL,the capacity of ·OH scavenging was 1.9650±0.0300 mg Vc/mL,T-AOC was 2.4368±0.0946 mg Vc/mL and the capacity of total reducing was 1.1283±0.0235 mg Vc/mL;With Trolox equivalent representation,the capacity of DPPH free radical scavenging of the pyroligneous acid was 1.07113±0.0201 mg Trolox/mL,the capacity of ABTS free radical scavenging was 2.7349±0.0206 mg Trolox/mL,the value of FRAP was 1.6852±0.0225 mg Trolox/mL,the capacity of ·OH scavenging was 4.8853±0.0550 mg Trolox/mL,T-AOC was 6.7052±0.3568 mg Trolox/mL and the capacity of total reducing was 2.5363±0.0570 mg Trolox/mL.(4)The method of pNPG was used to determine the inhibitory activity of pyroligneous acid on ?-glucosidase.The results showed that pyroligneous acid had significant inhibitory effect on ?-glucosidase.The inhibition ability was above 95% in the range of 2 to 8 times dilution.And its 50% inhibition dilution multiple(IC50)was 15.771±0.428.It means that the sampling amount of pyroligneous acid was 4.06±0.11 ?L.(5)The method of DNS was used to determine the inhibitory activity of pyroligneous acid on ?-amylase.The results showed that pyroligneous acid had inhibitory effect on ?-amylase.When the sampling quantity was larger than 250 ?L,the inhibition ability on ?-amylase activity was above 95% and its 50% inhibition sampling(IC50)was 136±2 ?L. |
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