Studies On The Characteristics Of Anthocyanin Accumulation And High Purity Preparation Technology Of Purple Buds From Tea Plants

Anthocyanins is a class 2-phenyl benzo pyran basic mother nucleus structure of plant secondary metabolites,antioxidant,scavenging free radicals and so on a variety of biological activity,is one of hotspots in the field of natural products,the development and utilization in recent years.The occurrence of chlorophyll in the buds of tea trees is mainly the anti-stress physiological manifestations of anthocyanin content caused by tea germ plasm,developmental characteristics and growth environment conditions.In traditional tea processing,tea products processed from purple sprouts are purple on the quality factors such as dry tea color,tea soup color and leaf bottom,and are considered to be unsuitable for processing fresh leaves of tea leaves and are mostly wasted.With deep research on the anthocyanins of the purple bud tea tree and the enhancement of health awareness,the high-value utilization of tea anthocyanins has attracted more and more attention in the industry.At present,the researches on anthocyanins in tea mainly focus on the breeding of purple bud tea cultivars,the regulation of anthocyanin metabolism,the preparation of crude extracts,the study of biological activity,the development of healthy products,etc,but the enrichment characteristics of anthocyanins in tea plants Mechanisms,anthocyanin component analysis methods,and preparation of anthocyanin high purity products have yet to be further explored.This study with Zixuan9803,Hongyafoshou and An73 three purple flower tea tree varieties?or lines?as the research object,the new tip in establishing a purple flower tea tree anthocyanins HPLC analysis method,on the basis of proven the purple flower tea tree anthocyanins enrichment characteristics,created the purple flower tea tree high anthocyanins refinement preparation technology,for the purple flower tea tree of anthocyanins provides scientific basis for the development and utilization of high value.The main results are as follows:The purple bud tea Zijuan,Zixuan9803,Hongyafoshou and An73 shoots were used as research objects.Using three plant anthocyanins?cyanidin-3-O-glucoside,pelargonin-3-glucoside,and delphinin-glucoside?as external standards,an HPLC method for simultaneous detection of various anthocyanins in purple bud tea tree was established.That is:it was extracted with 80%ethanol and refluxed,the tea is degreased with a 1:1:2 mixture of n-hexane:ethyl acetate:water.A C18 column was accompanied by acetonitrile and 0.2%phosphoric acid ultra-pure water as the mobile phase.Gradient,the organic phase elution gradient:0-55min,10%-40%,flow rate of 0.8 mL/min,column temperature 30°C,detection wavelength 520nm.Each anthocyanins can be effectively separated,The correlation coefficient of the standard curve was?0.99,The linear range is 5⁷5.00?g/L.The precision and repeatability of the method are good.The relative standard deviation of the stability?10%,The recovery rate is 75-88%.Seven major anthocyanin components in four kinds of purple bud tea samples were effectively separated.The method gets the characteristics of rapidity,sensitivity,accuracy,and low cost.At the same time,it is an analysis method that can be used to simultaneously detect and analyze various anthocyanin components in the purple bud tea.One bud and three leaves from three easy-purple bud tea varieties—HongYa FouFuShou,An 73 and ZiXuan 9803 were used as research materials.The effects of different tea varieties,different parts and different growing seasons on the enrichment characteristics of anthocyanins in purple bud tea plant were analyzed by UV-Vis spectroscopy and high performance liquid chromatography?HPLC?to provide reference to the high value utilization of purple tea buds and anthocyanins.The results showed that the effects of different tea varieties,different parts and different growing seasons on the enrichment of total anthocyanins have significant differences?P<0.05?.The enrichment capacity of different varieties showing a trend of ZiXuan 9803>An 73>HongYaFouShou,and different parts from strong to weak was the first leaf>the second leaf>bud>the third leaf.Besides,the seasonal changes of anthocyanin in three tea varieties with late spring,early summer,middle summer,late summer,early autumn and autumn showed the tendency of first increasing and then decreasing.The highest anthocyanins content of ZiXuan 9803 was 16.01mg/g and appeared in middle summer,An 73 was12.18mg/g and appeared in late summer,HongYaFouShou was 7.8mg/g and appeared in the early summer.The buds of Camellia sinensis cultivar?lines?were selected from the original bud of zixuan9803,and were successively degreased by acidic ethanol extraction and organic solvent extraction.After cation exchange resin decontamination,high-speed counter-current chromatography and preparative high performance liquid chromatography,5 were obtained.A high-purity tea tree violet buds anthocyanin component.To determine the optimum high-speed countercurrent solvent system ethyl acetate-n-butanol-acetonitrile-0.1%trifluoroacetic acid water?0.8:3.5:1.3:6,V/V?:and optimal preparative liquid chromatography conditions:C₁₈ column?20×250 mm,10?m?Preparative column,wavelength 280,mobile phase 1%acetic acid?A?-acetonitrile?B?gradient elution?0 to 55 min,10%to 40%B?,flow rate 5m L/min The amount is 0.8m L.Among them,the purity of the main unknown component 4 can reach 90%.According to the HPLC chromatograms of the literature and each component,it was presumed to be delphralate-3-O-?6-?E?-p-coumaric acid?pyran galactose Glycosides.