Studies On The Stability Of Anthocyanins Extracted From Blueberry And Purification Technology

As a functional beneficial material from plant,Anthocyanins,which are widely spreaded in tissues of varieties plants,are a group of flavonoid is composed with six kind of anthoyanidins combining with several monosaccharide or disaccharide and is of some biological effects,such as antioxidation,anticancer,free radical scavenger.At present,with studies on anthocyanin has been intensifying and people are more and more familiar to its properties at its every aspect, as well as its study achievements are increasingly accepected the aproval from medicine and coplasticizer, the anthocyanin resource get a rapid development. Although, the process of its development can't get a breakthrough with its development contrained by equippment, technology,the origin of raw materials and storage lacking of better mode as well as its own unstability. Now,domestic blueberry anthocyanin exploiting statuation at a low level in purity at a property of 25% yet, there also exists a situation that their purity just can reach 36% with a method of multiple speration using varieties of resins abroad. So there can have a lager blank in anthocyanin purification and speration for us to discuss. High speed count-current chromatography and high preparative performance liquid chromatography are applied to sperate blueberry anthocyanins,and building on its achievement we can undertake it strucrure identification and pharamcologic studies, and it will be a current of anthocyanin resouce develepment.In this study,blueberry juice concentration,which perchased from Shangzhi biological techology corperation,was used as a meterial. On base of full discussion the stability of blueberry anthocyanin,the resin AB-8, D101, LS300, LS300B, LS303, LS306, DM11, HP-20, HP2MGL absorbtion and desorbtion were tested, and the optimum resin was selected with its optimum process parameters were abtained. The high preparative performance liquid chromatography was applied to sperate blueberry anthocyanins. The main reseach results of this paper were obtained as follows:1.studies on stability of blueberry anthocyanin:①the stability of blueberry anthocyanin was tested with differe pH,the result shows the blueberry anthocyanin are very stable.and at pH3.0 the lost of blueberry anthocyanin is lower than any other pH. ②the stability of blueberry anthocyanin was tested under the temperature at 4℃, 25℃,40℃, and the result shows the blueberry anthocyanin will be very stable at 4℃.③pH,temperature,time orthogonal experiment was tested,the result shows the optuimun of blueberry anthocyanin stability is pH3.0,at temperature 4℃and 1.5h, and the sequence of the main factors of the effct to its stability is:temperature>pH> time.④the effection with different ion on blueberry anthocyanin stability:the result was discovered that oxidant Fe3++,Cu2+ have a sharp effect on blueberry anthocyanin stability. Reductant and K+ have some protection from its anthocyanin loss.2. Studies on separating of blueberry anthocyanin by macroporous resins:the material is blueberry concentrate juice, which has a anthocyanin purity of 1.24mg/g,Resin HP2MGL was selected from LS300, LS300B, LS303, LS306, DM11, D101, AB-8, HP-20, HP2MGL after comparing the absorption capacity at various pH and desorbtion effect with different concentration of alcohol.with a best Adsorbability 82.56%, desorpability 95.79%.The reslut of Static adsorption and desorption show that, the sequence of the main factors of the effct to separating of flavonoids from lotus leaves are:loading concentration> elution volume> elution flow rate>density of echohol>loading flow rate.An optuimum purification parameter with loading concentration 0,75g/ml,, ethanol concentration 50%, elution flow rate is 3.0 BV/h elution volume 6.0 BV,loading flow rate 3.5BV/h was built and applied to seperate blueberry anthocyanin.The result was obtained that purity is 28.9±0.3%, better than the report. And this method has an advantage of rapid,non-polution and non-toxic charactericsitc,and will has some good reference value for its industrical production.3. Studies on purification of blueberry anthocyanin by HSCCC:Under the conditions that solvent system is n-butanol-TBME-acetonitrile-0.2%TFA(1.5:4:1: 5), the rotational speed is 850 r/min, flow rate of immobilityphase is 20 mL/min, flow rate of mobile phase is 2.0 mL/min, detection wavelength is at wavelength of 280 nm, and three peak was seperated with three kind of meterial seperated.