Ultra-high Pressure Extraction Of Anthocyanins From ‘shuanghong' Vitis Amurensis Rupr,and Its Purification,component Analysis And The Development Of Anthocyanins Beverage

Vitis amurensis Rupr belonged to deciduous vine and mainly are planted in the northeast of China and inner mongolia area,they have characterized by strong cold-resistance and high anthocyanins content,they are widely used to brew wine,having the main varieties of ‘Shuanghong',‘Beibinghong',‘Shuangyou' and etc.‘Shuanghong' vitis amurensis was a hybrid with Tonghua No.3 as female parent and Shuangqing as male parent.With the increasing emphasis on health and owning to a variety of physiological functions,anthocyanins have attracted consumers' attention and love,therefore,extracting anthocyanins from ‘Shuanghong' vitis amurensis and developing new products are of great significance to the market and anthocyanins industry.Using ‘Shuanghong' vitis amurensis as raw material,in this study,ultra-high pressure technique and ethanol extraction method were applied for the extraction of anthocyanins,the anthocyanins were separated and purified by D101 macroporous resin,high performance liquid chromatography(HPLC)was used to analyze the components of anthocyanins.Finallly,we made the vitis amurensis juice beverage rich in anthocyanins with the grape juice and white sugar as raw material,the purified freeze-dried powder of anthocyanin and citric acid as auxiliary materials through adjusting and ultra-high pressure sterilization,the results were as follows:1.Ultra-high pressure technique was applied for the extraction of anthocyanins from ‘Shuanghong' vitis amurensis,the single factor tests were applied to inspect the effects of liquid ratio,pressure and pressure-holding time on the yield of anthocyanins,the response surface methodology was applied to optimize the extraction conditions.The results showed that the optimal extraction processing parameters were liquid/material ratio of 8.5:1,extraction pressure of 200 MPa and pressure treatment time of 2 min,under these conditions,the highest yield of anthocyanins was 0.304±0.007 g/100 g.2.Heat extraction method was applied for the extraction of anthocyanins from ‘Shuanghong' vitis amurensis,the single factor tests were applied to inspect the effects of liquid ratio,extraction temperature,extraction time on the yield of anthocyanins,the response surface methodology was applied to optimize the extraction conditions.The results showed that the optimal extraction processing parameters were liquid/material ratio of 5.5:1,extraction temperature of 55 ? and extraction time of 30 min,under these conditions,the highest yield of anthocyanins was 0.240±0.005 g/100 g.3.The results of comparison of two extraction methods were as follows:extraction yield of anthocyanins by ultra high pressure was 26.7%,higher than that obtained with the traditional heat extraction method and with short extraction time,the results showed that the extraction of anthocyanins by ultra high pressure was better.4.Five different macroporous resins(NKA-?,NKA-9,AB-8,D-101,HPD-100)were evaluated for their use to purify anthocyanins from ‘Shuanghong' vitis amurensis.Macroporous resin D101 was selected as the most suitable one by using static adsorption and desorption experiments and its optimum adsorption desorption temperature were 25 ?.The best dynamic adsorption and desorption parameters using D101 macroporous resin were as follows: the temperature was 25 ?,sample concentration of 22.5 mg/mL,sample loading volume of 8 BV(resin bed volume)the adsorption speed of 2 mL/min,the optimal concentration of eluent was 75 % ethanol-0.01 % hydrochloric acid,the elution flow rate was 2 mL/min,the desorption solution was 7 BV.Under these conditions,the purity of anthocyanins from ‘Shuanghong' vitis amurensis was 27.78±0.043 %,higher than the purity of anthocyanins products(25 %)on the market.The results showed that D101 macroporous resin had better purification effect on anthocyanins from ‘Shuanghong' vitis amurensis.5.HPLC method was used to analysis the components of anthocyanins from ‘Shuanghong' vitis amurensis,the results showed that the anthocyanins in ‘Shuanghong' vitis amurensis included five categories,the content of anthocyanins were ranked from high to low as follows: malvidin 99.91 mg/100 g,delphinidin 51.03 mg/100 g,cyanidin 19.30 mg/100 g,petunidin 13.54 mg/100 g,peonidin 3.00 mg/100 g.6.The vitis amurensis juice and white sugar were used as raw materials,the citric acid and purified freeze-dried powder of anthocyanins were used as auxiliary materials to get the vitis amurensis juice beverage rich in anthocyanins,the single factor tests and the orthogonal test were applied to determine the best formula of vitis amurensis anthocyanins beverage as follows: 20 % of vitis amurensis juice,13 % of soluble solids,0.23 % of freeze-dried powder of anthocyanins,0.34 % of total acid.The beverage was deep rose red,sweet and sour taste,with the unique aroma of grape and the best quality,the anthocyanins content was 80.31 mg/100 mL.7.In order to study the effect of different sterilization methods on the quality of beverage,and according to the best formulation,anthocyanin beverage was sterilized using the method of water bath sterilization and ultra-high pressure sterilization.The sensory score,the total number of colonies,color and anthocyanins retention rate were as evaluation index and the optimal sterilization conditions of ‘Shuanghong' anthocyanins beverage were determined.Under the condition of sterilization,the beverage was placed at 4? and 37?to do shelf life test.Using full trademark,half trademark and naked trademark to packaged beverage,the best packaging of beverage was determined through comparing the anthocyanins retention rate.The results showed that the sensory score was 97.83±0.29 and the preservation rate of anthocyanins was 95.63±1.15 %,the total bacteria was 0 CFU/mL in the beverage,which accorded with the national standard after 450 MPa ultra-high pressure sterilization for 13 min,the color difference was small before and after ultra-high pressure processing,which showed that the original color of the beverage was maintained after 450 MPa ultra-high pressure sterilization for 13 min.The preservation rate of anthocyanins using full trademark declined slowest with the extension of time,which was beneficial to the preservation of anthocyanins,the quality guarantee period of ‘Shuanghong' anthocyanins beverage could last as long as 270 days.