| Blueberry, also known as bilberry, basic nutrition of which are fructose, protein, fat, amino acids, pterostilbene, superoxide dismutase, rich in anthocyanins. Blueberry anthocyanins have hypoglycemic, anti-inflammatory, anti-tumor, proliferation and absorption rate is high in the small intestine. Majiang of Guizhou province introduced blueberries from the beginning of the 2000, then, establishment of a 13.33 hm2 demonstration park in 2003. To 2008, a total of 30 kinds of blueberry were introduced from 4 major varieties. But there are less research of Guizhou blueberry, this paper mainly identificate kinds of anthocyanin and investigate the process for extraction and purification of Majiang rabbit eye.Furturemore, In vitro antioxidant activity of anthocyanins from different procedure was determined. The innovation points are showing as bleow:1. In order to gain the extraction rate of the Blueberry anthocyanin, the single factor experiment and the response surface Box-Behnken test were designed to investigate the effect of different acid, temperature, time, ethanol volume fraction, ratio of solid-to-solvent, extraction times with Ultrasound-assisted Ethanol Extraction Technology. The optimal extraction conditions of blueberry anthocyanins were extracted 3 times at 40℃, and ultrasonic time was 20 min. The ratio of material to liquid was 1:6. The solvent was 95% ethanol which contains 0.3% HCl(V/V). Finally, we got 187.92 mg anthocyanins 100 g blueberry. The 15 m L concentrate was extracted 4 times with 30 m L ethyl acetate, and the content of anthocyanins decrease from 187.92 mg/100 g to 135.32 mg/100 g blueberry. The AB-8 macroporous resin show stronger adsorption rate and desorption rate. The 15 m L concentrate which extracted with ethyl acetate was absolutely absorbed at 1.5 h. Then, used 25 m L distilled water to remove fructose. Finally, the 70. m L 95% ethanol which contained 0.3% HCl( V/V) was used to elute anthocyanins. The content of blueberry anthocyanins decreases from 135.32 mg/100 g to 95.83 mg/100 g.2. A systematic method for anthocyanin identification using tandems mass spectrometry(MS/MS) coupled to high-performance liquid chro-matography(HPLC) was developed. Scan for the precursor ions of commonly found anthocyanidins(cyanidin, delphinidin, malvidin, pelargonidin, petunidin, and peonidin) using LC/MS/MS on a triple quadrupole instrument allows for the specific determination of each category of anthocyanins. Further characterization of each anthocyanin was performed using MS/MS selected reaction monitoring(SRM). ESI/MS/MS coupled with HPLC-PDA identificated 18 anthocyanins of Majiang Rabbit eyes blueberries( Gardenblue), of which delphinidin-3-galactoside, delphinidin-3-glucoside, cyanidin-3-galactoside, delphinidin-3-arabinoside, cyanidin-3-glucoside, cyanidin-3-arabinoside, petunidin-3-glucoside, peonidin-3-galactoside, petunidin-3-arabinoside, peonidin-3-glucoside, malvidin-3-galactoside, peonidin-3-arabinoside, malvidin-3-glucoside, cyanidin, malvidin-3-arabinoside, petunidin, Peonidin, malvidin, respectively. Across this genotypes( Gardenblue), the malvidin type constituted 36.94% of the total anthocyanins, where the proportions of cyanidin and delphinidin were 26.76%, 16.69%, respectively. The highest content of individual anthocyanin was the malvidin-3-glucoside(15.86%). However, the commercial blueberry just contains four individual anthocyanins: cyanidin-3-glucoside, peonidin-3-glucoside, petunidin, cyanidin.3. Anthocyanin(λmax = 470–580) might interfere with the absorbance of DPPH?(λmax = 515 nm) in the colorimetric method leading to the failure in detection of small changes in the DPPH? absorbance. HPLC method was specific for DPPH? and was based on the reduction in DPPH? peak area according to different rention time. The antioxidant activity blueberry was achieved by HPLC method, in terms of 1,1-diphenyl-2-picrylhydrazyl radical(DPPH?) scavenging capacity was evaluated. Subsequently, the predominant active anthocyanins were screened from Majinag rabbit eye(Gardenblue) was : Cyanidin, Petunidin, Peonidin, Malvidin. The decreases of PAs ranged from high of delphinidin-3-arabinoside(22.38%), Delphinidin-3-galactoside(21.62%), Delphinidin-3-glucoside(22.24%), to a low of 4.04% for Cyanidin-3-glucoside. The results indicated that Majinag blueberry(Gardenblue) show stronger antioxidant activities than commercial blueberry. The DPPH? free radical scavenging rate of Majinag rabbit eye blueberry(Gardenblue) reached to a top of 85.74% at 1 mg/m L, while the commercial blueberry was 80.64% at 1.5 mg/m L. After the crude of Majinag rabbit eye blueberry(Gardenblue) exetracted by ethyl acetate, the DPPH? free radical scavenging rate increase from 75.27% to 84.93%. However, it drops to 74.16% when purified by AB-8 macroporous resin. |
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