摘要
为建立一种快速高效分离乳清蛋白胃蛋白酶水解液中的抗氧化肽的方法,采用基于有机溶剂-盐的双水相体系进行分离。以上相多肽提取率和ABTS自由基清除率为评价指标,结合反相-高效液相色谱(RP-HPLC)检测,探究多肽的分配规律,确定最优分离条件。对上、下相和水解液中多肽进行纯化,分别测定其ABTS自由基清除能力、DPPH自由基清除能力、羟自由基清除能力以及还原能力,并用基质辅助激光解吸飞行时间质谱(MALDI-TOF)和液相色谱对纯化后多肽进行表征。结果表明:双水相体系分离抗氧化肽的最优条件为加入4mL 25%NaH_2PO_4,3 mL正丙醇,NaCl 0.75 g/10 m L,此时,大多数的抗氧化性多肽被分配到双水相上相中,且纯化后上相多肽各项抗氧化值均高于下相及水解液多肽。结论:亲水有机溶剂-盐双水相体系可对水解液中抗氧化性多肽进行分离富集,且效果良好。
To explore a new and simple rapid method for the separation of antioxidant peptides from pepsin-catalyzed hydrolysis of whey protein isolate（WPH）, an aqueous two phase systems（ATPS） formed by organic solvents and salts was developed. The effect of component in ATPS on the separation was evaluated by the extraction yield, 2, 2＇-Azinobis-（3-ethylbenzthiazoline-6-sulphonate）（ABTS） radical scavenging capacity combined with Reversed Phase Highperformance Liquid Chromatography（RP-HPLC）. Then the peptides in top phase, bottom phase and WPH were purified to determine the ABTS radical scavenging capacity, DPPH radical scavenging capacity,·OH radical scavenging capacity and ferric ion reducing antioxidant power. RP-HPLC and matrix assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF） were used for characterization. The obtained data indicated that the antioxidant peptides were preferentially partitioned into the top phase and the ATPS composed of 4 mL 25% sodium dihydrogen phosphate, 3 mL1-propanol, and added sodium chloride 0.75 g/10 mL showed good selectivity on the antioxidant peptides. Under this condition, the most antioxidant peptides with high oxidative stability was separated to the top phase, and the antioxidant activity of the purified top phase peptides was higher than that of the bottom phase and the WPH. The representation by RP-HPLC and MALDI-TOF also proof it. Conclusion： ATPS composed of organic solvents and salts could extract and enrich antioxidant peptides to the top phase from WPH.
作者
冯志彪
屈玉霄
张晓庆
刘春红
姜彬
Feng Zhibiao;Qu Yuxiao;Zhang Xiaoqing;Liu Chunhong;Jiang Bin(College of Science,Northeast Agricultural University,Harbin 150030)
出处
《中国食品学报》
EI
CAS
CSCD
北大核心
2018年第10期170-177,共8页
Journal of Chinese Institute Of Food Science and Technology
基金
国家自然科学基金项目(31201366)
哈尔滨市科技创新人才项目(2016RAQXJ052)
