Study On Extraction, Purification And Functional Characteristic Of Blueberry Anthocyanins

Blueberry is abundant and distributing over the country.Anthocyanins have been discussed in relation to a wide range of function characteristic such as vision improvement, antioxidant,antidecrepitude,antiulcer,antivirus and anticancer and so on.In this paper, Saintcloud blueberry with most anthocyanin content was selected after comparing anthocyanin content of 4 kinds of blueberry fruits.Anthocyanins were systematically studied from Saintcloud blueberry as material.It is very important to understand its bioactivities and use the blueberry resource.In this paper,anthocyanins were extracted from Saitcloud blueberry fruits.The optimum industrial process of extraction of blueberry anthocyanins were determined by solvent and enzyme method.Blueberry anthocyanins' conditions of absorption and desorption which were static and dynamic were studied by AB-8 macroporous resin.Blueberry anthocyanins were purified by polyamide and silica-gel column chromatography.Blueberry anthocyanins were separated by thin-layer chromatography.Constitutes and structrue of blueberry anthocyanins was identified by RP-HPLC/MS.The stability of blueberry anthocyanins were tested.Microencapsulation and molecular modification of blueberry anthoeyanins were studied.The effect of blueberry anthocyanins on antioxidant was investigated by the method of chemical simulation in vitro.The effects of anthocyanions from blueberry fruits on hypolipidemic and antioxidative in hyperlipidemic rats were studied by animal tests.The main results were as follows:1.The optimum extraction technology parameters determined by ethanol were as follows: 60%ethanol,material-solvent ratio 1g:15mL,extraction time 120min,pH value 3.0, extraction temperature 40℃and extraction 2 times.Anthocyanins content was 328.91 mg/100g fresh fruits.2.The effect of cellulase,pectinase and their mixture on blueberry anthocyanins extraction was studied.The result showed that cellulose could enhance the extraction content of anthocyanins,but pectinse could not.In addition,the extraction content of anthocyanins by their mixture was lower than by cellulase.The optimum parameters of extraction technology by cellulase were enzymatic quantify 5mg/g,material-liquid ratio 1 g:8mL,enzymolysis time 60min,pH value 5.0,enzymolysis temperature 45℃and extraction 2 times.Blueberry anthocyanins content was more 49.53%by cellulase on extraction than water.3.Among 10 kinds of macroporous resins,AB-8 resin has better capabilities of absorption and desorption to purify crude blueberry anthocyanins.The conditions of static absorption and desorption of blueberry anthocyanins were that anthocyanins concentration in balance was 750mg/L,pH value of absorption was 3.0,the elution solvent was pH3.0 and 60%ethanol.The optimum conditions of dynamic absorption and desorption of blueberry anthocyanins were as follows:flow rate of absorption and desorption 1mL/min,concentration 3.0mg/mL.The elution solvent was 3BV(bed volume)60%ethanol.In those conditions,the anthocyanins produced by this technological process was purplish dark powder.Blueberry anthocyanins were purified by silica-gel and polyamide column chromatography after AB-8 resin purifying,the colour value was 79.20,85.30 respectively.Anthocyanins were separated by thin-layer chromatography.Separation was well done by butanol-acetic-water(4:1:5)and water-ethanol- butanol - acetic ester(12:8:24:36) as developing solvent.Two kinds of anthocyanins were isolated by preparative HPLC.Sample 1 was Malvidin3-arabinoside, Sample 2 was Malvidin3-glucoside by UV,IR,MS and NMR.4.The wavelength of absorption respectively was 520nm and 535nm in pH3.0 citric acid solvent and methanol.Saintcloud blueberry was found to contain 17 anthocyanins by HPLC/MS.The 17 of them were tentatively identified as Peonidin3-galactoside, Delphinidin3-galactosid,Cyanidin3-galactoside,Cyanidin3-glucoside,Petunidin3-galactoside ,Petunidin3-glucoside,Cyanidin3-arabinoside,Peonidin3-glucoside,Malvidin3-galactoside,Mal -vidin3-glucoside,Malvidin3-galactoside,Petunidin3-arabinoside,Delphinidin3-glucoside,Peon -idin3-(6"-acetyl)galactoside,Malvidin3-(6"-acetyl)galactoside,Peonidin3-(6"-acetyl) glucoside,Malvidin3-(6"-acetyl)glucoside.5.Blueberry anthocyanins are water-soluble pigments.The anthocyanins were stable under pH 3.0,below 60℃.It was heat-resistant to high temperature in a short time.The anthocyanins were sensitive to light and stable to microwave.It was found that oxidizer H₂O₂, reductive Na₂SO₃ and ascorbic acid could decrease the stability of anthocyanins and result in its more rapid discoloration.Sucrose and glucose had no harmful effect on the stability of blueberry anthocyanins,but had certain color maintenance at higher concentration.Sodium benzoate and K⁺ had no effect on the stability of anthocyanins.Mg²⁺,Ca²⁺,Cu²⁺and A1³⁺ had a certain effect of clout improvement,but no effect on the stability of anthocyanins.Na⁺,Zn²⁺ and Mn²⁺could not holy strengthen the colour at higher concentration,but enhance the stability of blueberry anthocyanins.Fe²⁺,Fe³⁺ and Pb²⁺ were harmful to the stability of blueberry anthocyanins while Fe³⁺ and Pb²⁺ could lead to precipitate in blueberry anthocyanins solution.6.The optimum technology parameters by using arabic gum as wall materials were as follows:the proportion of anthocyanins to wall materials was 3:197,emulsion concentration was 20%,inlet air and outlet air temperature were 180℃and 70℃respectively.The optimum technology parameters by using mixture ofβ-cyclodextrin,maltodextrinn,and arabic gum as wall materials were as follows:the ratio of core mateials to wall mateials 3:97,β-cyclodextfinto maltodextriun 1:2,arabic gum 7.5%,emulsion concentration 20%.The optimum technology parameters with mixture of soybean protein isolatidn(SPI) and maltodextrin showed that SPI and maltodextrin was in proportion of 3:7 as the materials,the proportion of core mateials to wall mateials was 4:96,the emulsion concentration was 15%(W/V).The optimum technology parameters by using ethyl cellulose as wall materials were as follows:EPG concentration was 15%,the proportion of anthocyanins to wall materials was 1:19,emulsion concentration was 4%respectively.Anthocyanins of microencapsulation have higher releasing rate in vitro.Releasing rate could exceed 90%after 60min in gastric juice of simulation.The temperature and light stability of microencapsulation and anthocyanins of molecular modification could be improved.7.Blueberry anthocyanins could inhibit peroxidation of lard and lipid,and have stronger effect of lipid peroxidation than ascorbic acids.Blueberry anthocyanins have the reduction ability,and the positive correlation existed between reductive ability and concentration. Blueberry anthoeyanins could effectively scavenge hydroxyl free radicals,superoxide anions, DPPH and peroxide.8.The serum TG,TC,LDL-C concentration,ApoB content and atherosclerosis index in rats fed with anthocyanins from blueberry fruits(ABBF)were significantly lower than those in model group,while serum HDL-C concentration and ApoA I content were higher.Comparing with model group,T-AOC,SOD,GSH-Px were increased significantly in serum and liver by feeding with ABBF,while MDA content were decreased in serum and liver.ABBF could decrease blood lipids level,improve blood lipid metabolism,inhibit lipid peroxidation,inhibit formation and development of antherosclerosis.