The Extraction And Purification Of Cyclic Adenosine Phosphate From The Jujube

China is quite rich in jujube,whose planting area and output account for 99% of the area and production in the world.Jujube contains abundant nutrients,such as polysaccharide,polyphenol compounds,organic acids,alkaloids,flavonoids,cycle adenosine phosphate,etc.Jujube possess a variety of health care function,in addition to protecting liver,inhibiting cancer,and antioxidant.This paper was mainly carried out to detect,extract and purify cAMP in ZanHuang jujube.Jujube concentrated solution of cAMP was obtained by fermentation method.This study provided a new direction for Hebei ZanHuang jujube processing,improved the comprehensive economic benefits of the jujube,and drove the jujube industry economy in Hebei.The cAMP detection method was established by HPLC.Phosphate-methanol elution system was adopted with gradient elution way and the UV detection wavelength was259 nm,The HPLC Separation column was ODS column.The flow rate was 0.6 mL/min.The standard curve of cAMP was y = 79786 x.The correlation coefficient was 0.9996,and the accuracy of this method and the stability of the RSD value was less than 1%.The quantitative of limited was identified as 0.039 mg/L.This paper respectively adopted ultrasonic assisted extraction,microwave assisted extraction,enzymatic extraction and hot water extraction methods.By comparing the results analysis,hot water extraction method was used.For extracting time,temperature,material liquid ratio runing single factor experiments,According to the results of single factor experiment analysis,designed three factors three levels orthogonal experiment,According to the results of orthogonal experiment,Finally the optimum extraction conditions for cAMP : extraction temperature was 70℃,extraction time was 1.5 h,extraction of material liquid ratio was 1:20.Under optimal extraction conditions taking extraction of series of experiments,Determining the extraction of jujube was leaching at a time.CAMP extraction rate reached 94%.The extraction process was completed.Preliminary purification used macroporous resin.Through the choice of macroporous resin type,DA-1 type macroporous resin was finally determined,and studied the macroporous resin regeneration.Results showed that DA-1 type macroporous resin possessed a good reproducibility and could be reused.In addition,to determine eluent,acid solution,alkali solution,30% ethanol solution were experimented respectively and finally,30% ethanol solution was determined.the liquid ofsample and quantity of resin was carried out in accordance with the 5:1 loading,the loading number for 3 times.Preliminary purification could remove most of the sugar and total phenolics,and cAMP sample was obtained with the purity 5.18%.The preliminary purification eluent of macroporous resin was conducted with decolorization,such as alkali extraction and acid precipitation,PVPP adsorptive decolorization,diatomite load chitosan decoloring method,copper ion-ammonia decoloring method,polyphenol protein complex decoloring,activated carbon adsorption decolorizing,and alumina column decoloring methods.Eventually,alumina decoloring was determined as the decolorization method.Through the alumina decoloring,the color of the solution turned from yellow to colorless.The purity of cAMP was 37.33%.To further purify the sample,silica gel column was adopted and 0.4 g solid sample was loaded into the silica gel column with the methanol as elution solution and the flow rate 0.1 ml/min.Finally purity of 2 mL cAMP samples collected reached 50.87% by fractional collections methods.Sugar of cAMP extracs from jujube was removed by the fermentation method and the concentration of cAMP reached 800-1000 mg/L by enrichment.