紫象草原花青素的大孔吸附树脂纯化、平均聚合度及体外抗氧化活性测定  

Purification Process by Macroporous Sorbent Resin,Determination of Average Polymerization and In Vitro Antioxidant Activity of Proanthocyanidins from Pennisetum purpureum Schumab

作　　者：冉思蛟 王元龙 倪海花 刘赛格 瞿秋涵 康馨月 高彦华 RAN Sijiao;WANG Yuanlong;NI Haihua;LIU Saige;QU Qiuhan;KANG Xinyue;GAO Yanhua(College of Animal and Veterinary Sciences,Key Laboratory of Animal Science of National Ethnic Affairs Commission of China,Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resources Reservation and Utilization,Southwest Minzu University,Sichuan Chengdu 610041,China)

机构地区：[1]西南民族大学畜牧兽医学院,动物科学国家民委重点实验室,青藏高原动物遗传资源保护与利用教育部重点实验室,四川成都610041

出　　处：《饲料工业》2024年第3期72-81,共10页Feed Industry

基　　金：四川省自然科学基金项目[2023NSFSC1148];2021年度西南民族大学教育教学研究与改革重点项目[2021ZD63];西南民族大学中央高校基本科研业务费项目[ZYN2023104]。

摘　　要：试验旨在利用大孔吸附树脂获得紫象草原花青素粗提物的纯化条件,并探究在此条件下获得纯化物的平均聚合度和体外抗氧化活性。试验采用单因素试验设计,通过比较3种大孔吸附树脂D101、HP-20、ADS-17的吸附与解吸特性,选择最优吸附树脂类型,对紫象草原花青素粗提物进行洗脱浓度、上样体积和洗脱体积的筛选,之后利用静态和动态吸附与解吸试验确定纯化条件,制备紫象草原花青素纯化物;采用盐酸-香草醛法测定纯化物原花青素的平均聚合度及其对1,1-二苯基-2三硝基肼(DPPH·)和羟自由基(·OH)的体外清除率。结果表明:采用φ2.6 cm×30.0 cm玻璃层析柱,选择D101型大孔吸附树脂、90%(V/V)浓度乙醇为洗脱剂,上样体积480 mL,洗脱体积155 mL时纯化效果较好,纯化后原花青素B2含量由纯化前10.01 mg/g提升至13.82 mg/g,提升了36.06%;紫象草原花青素粗提物经乙酸乙酯萃取后,获得水相和乙酸乙酯相的平均聚合度分别为26.87、13.63,纯化物的平均聚合度为14.67。当紫象草原花青素纯化物质量浓度为100 mg/mL时,DPPH·、·OH清除能力分别为99.55%、97.72%,相当于1 mg/mL维生素C,并且紫象草原花青素纯化物体外抗氧化活性显著高于粗提物(P<0.05)。综上所述,试验获得了大孔吸附树脂纯化紫象草原花青素的条件,纯化处理基本不影响原花青素提取物的平均聚合度,并发现原花青素纯化物具有良好的体外抗氧化活性。This experiment aimed to optimize the purification conditions of crude extracts of proanthocyanidins from Pennisetum purpureum Schumab using macroporous sorbent resin,and to investigate the average polymerization degree and in vitro antioxidant activity of the purified products obtained under these conditions.A single-factor experimental design was used to compare the adsorption and desorption characteristics of three types of macroporous adsorbent resins D101,HP-20 and ADS-17,and to select the optimal adsorbent resin type for the elution concentration,loading volume and elution volume of the crude extract of Pennisetum purpureum Schumab proanthocyanidins,followed by static and dynamic adsorption and desorption tests to determine the purification conditions and prepare the purified Pennisetum purpureum Schumab proanthocyanidins.The average polymerization degree of the purified proanthocyanidins and their in vitro scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH·)and hydroxyl(·OH)radicals were determined by the hydrochloric acid-vanillin method.The results showed that the purification effect was better when the sample volume was 480 mL and the elution volume was 155 mL,and the content of proanthocyanidin B2 was increased from 10.01 mg/g to 13.82 mg/g after purification by 36.06% on a φ2.6 cm×30.0 cm glass chromatography column with D101 type macroporous adsorbent resin and 90%(V/V)concentration ethanol as eluent.The average polymerization degree of aqueous phase and ethyl acetate phase were 26.87 and 13.63,respectively,and the average polymerization degree of purified product was 14.67.When the mass concentration of pennisetum purpureum Schumab proanthocyanidins purified product was 100 mg/mL,the scavenging ability of DPPH·and·OH radicals was 99.55% and 97.72%,respectively,which was equivalent to 1 The antioxidant activity of the purified Pennisetum purpureum Schumab proanthocyanidins was significantly higher than that of the crude extract(P<0.05).In conclusion,the conditions for purification o

关 键 词：紫象草 原花青素 大孔吸附树脂 平均聚合度 体外抗氧化活性 

分 类 号：S816.32[农业科学—饲料科学]