Study On Distilling Pineleaf Essential Oil And Puerarea-Flavonoid With Enzymatic Hydrolysis

The present experiment was designed to study the effect of enzymatic hydrolysis on distilling and compare the substance distilled of common way and enzymatic hydrolysis by chromatography.Pineleaf and puerarea was hydrolyzed by multi-component enzymes consisted of Cellulase, Protease and Pectinase, which have very high activity and pineleaf essential oil and puerarea-flavonoid were distilled by general way also. The distilling rate was chosen as index to optimize the condition parameter of multi-component enzyme hydrolysis with single factor and orthogonal design,and the optimized group were obtained as following: temperature 50℃, pH value 4.5, time 6h, and additive quantity 4% .The results showed that the distilling rate of pineleaf essential oil and puerarea-flavonoid could be enhanced by 37.9% and 26.3% with the optimized hydrolysis condition parameter. The difference between two distilling ways is very prominent according T Student analysis (P<0.001). It indicated that the distilling rate of pineleaf essential oil and puerarea-flavonoid by multi-component enzyme was increased significantly in comparision with general way.Pineleaf and puerarea was hydrolysed by Aspergillus niger AN02, which produces Cellulase, Protease and Pectinase, which have very high activity and pineleaf essential oil and puerarea-flavonoid were distilled by general way also. The distilling rate was chosen as index to optimize the condition parameter of Aspergillus niger AN02 hydrolysis with single factor and orthogonal design,and the optimized group were obtained as following: temperature 40℃, pH value 4.5, time 6h, additive quantity 6%. The results showed that the distilling rate of pineleaf essential oil and puerarea-flavonoid could be enhanced by 34.3% and 21.1% with the optimized hydrolysis condition parameter. The distinction between two distilling ways is very prominent according T Student analysis (P<0.01). The results suggested that distilling rate of pineleaf essential oil and puerarea-flavonoid are increased prominently by Aspergillus niger AN02 at the optimal hydrolysis condition and that distillation by Aspergillus niger AN02 were more economic and easy than distillation by multi-component enzyme.The pineleaf essential oil and puerarea-flavonoid distilled were identified by chromatography. The results manifested that the TLC of pineleaf essential oil and puerarea-flavonoid distilled with hydrolysis and common way have no difference basically and that the layer dots of distillage and standard sample were lie in the same level. The results of Gas Chromatography suggested that some big wave crests in the GC of pine-oil standard sample are also appeared in the others GC at synchronization and that the three GC figures of distillage are resemblant in the whole. The results of High Performance Liquid Chromatography showed that the most wave crests in the HPLC of puerarea-flavonoid standard sample is also appeared in the others HPLC at synchronization and that the OD number of the most wave crests in the four figure of HPLC among 700 to 800 mAU. According to the results of chromatography, the distillages have same elements in comparision with the standard sample and the elements of distillages after hydrolysed have not been changed.