Studies On Separation Arid Purification For CAMP From Ziziphus Jujube

In this research, the dry and crushed jujube is selected as raw material. Researching on thecAMP(Cyclic adenosine-3’,5’-monophosphate) is analyzed by UPLC (Ultra Performance LiquidChromatography) method and the cAMP is extracted by the ultrasound extraction technology; researchingon the technology of efficient enzymatic hydrolysis and release cAMP technology and the method of theseparation and purification of cAMP. The results are as follow:1. The cAMP is analyzed by UPLC (Ultra Performance Liquid Cromatography) method. In this method,pH2-11，ZORBAX Extend C182.1×100mm1.8-Micron Rapid Resolution HD2.1×100mm1.8-Micron isused. The mobile phase is as follows: V(methanol):V(0.1mol/L phosphate buffer)=20:80, and a flow rate of1.0mL/min, temperature of column was25℃, quantity of injection is10uL, the ultraviolet detector wavelength is258nm. The result shows that the linear relation of the cAMP standard curve is good. The methodis simple, sensitive, and reproducible. It can rapidly determine the cAMP from Ziziphus jujube.2. The best ultrasound extraction technology of cAMP from Ziziphus jujuba Mill was researched. Singlefactor experiments to determine the parameter changes in the level range, ratio of solvents and material, theultrasonic parameters of time, the treatment mode, the ultrasonic power as independent variables tooptimize the extraction process parameters for cAMP from jujube by using central composite design andresponse surface analysis. The results showed that the optimum extraction conditions were ratio of solventsand material1:15.5, the ultrasonic power314W, the ultrasonic parameters of time11.7min, the treatmentmode1.08min. Under the optimum extraction conditions, the measured value was502.2ug/g, degree ofagreement reached96.80%.3. The polysaccharide from Ziziphus jujube has a great influence on extraction yield of cAMP. Theextraction yield of cAMP reaches the maximum value and then drops down. The liquid pH stability rangesfrom pH4.24to pH4.18.4. The jujube pigment and polysaccharide are absorbed by using AB-8macro-porous resin. The sampleliquid pH4, the sample liquid concentration20ug/mL, the sample liquid velocity2mL/min, the maximumsample liquid volume300mL.40%ethanol solution is selected as elution solution. The flow speedof elution2mL/min, the maximum elution solution volume160mL. The color of elution solutionis near colorless.5. The cAMP from Ziziphus jujube is separated and purificated by using D-290macro-porous resin.Single factor experiments to determine the parameter changes in the level range, the sample liquidconcentration, the sample liquid speed, concentration of eluent, speed of eluent as independent variables tooptimize the separation and purification process parameters for cAMP from jujube. The result shows thatthe optimum purification conditions are the sample liquid concentration400ug/mL, the sample liquidspeed2.5BV/h, concentration of eluent (0.1mol/L)HCl-(0.4mol/L)NH4Cl, speed of eluent2.5BV/h.Under the optimum purification conditions, the measured value was212.38ug/g, the purification rate is42.29%.