食品科学

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O2 - ·荧光探针法检测及其应用

赵永强,林 洪,李来好,杨贤庆,郝淑贤,张牧天   

  1. 1.中国海洋大学食品科学与工程学院,山东 青岛 266003;2.中国水产科学研究院南海水产研究所,农业部水产品加工重点实验室,国家水产品加工技术研发中心,广东 广州 510300;
    3.北京师范大学-香港浸会大学联合国际学院,广东 珠海 519085
  • 出版日期:2013-06-25 发布日期:2013-06-17

Development and Application of Fluorescence Probe Method for Superoxide Anion Radical Detection

ZHAO Yong-qiang,LIN Hong,LI Lai-hao,YANG Xian-qing,HAO Shu-xian,ZHANG Mu-tian   

  1. 1. College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China;
    2. Key Laboratory of Aquatic Product Processing, Ministry of Agriculture, National R & D Center for Aquatic Product Processing,South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510300, China;
    3. Beijing Normal University-Hong Kong Baptist University United International College, Zhuhai 519085, China
  • Online:2013-06-25 Published:2013-06-17

摘要:

以2-氨基吡啶与吡啶-2-甲醛为原料,经亲核取代反应合成了一种新型荧光探针——2-(2’-吡啶亚胺甲基)吡啶(2-APC),所得目标产物经熔点测定、元素分析、红外光谱与核磁共振波谱等方法表征确认。利用2-APC与超氧阴离子自由基(O2-·)发生荧光淬灭反应的原理,建立了一种测定邻苯三酚自氧化体系产生O2-·的方法,该方法反应体系最佳条件为反应pH8.2、反应温度(T)40℃、反应时间(t)40min,测定条件为:λex=295nm、λem=365nm,狭缝宽度5nm。邻苯三酚在0.4×10-6~8.0×10-6mol/L浓度范围内与相对荧光强度呈良好线性关系,线性回归方程为y=37.567x+55.581,R2=0.9834。应用该方法对L-抗坏血酸清除O2-·能力进行评价,结果表明L-抗坏血酸清除O2-·的IC50值为0.182mmol/L。

关键词: O2-·, 荧光探针, 合成, 应用

Abstract:

A novel fluorescence probe 2-APC was synthesized based on nucleophilic substitution reaction of 2-aminopyridine
and 2-pyridinecarbaldehyde in this study. The reaction product was characterization by melting test, elemental analysis,
infrared spectrum and 1HNMR. A fluorescence probe method for superoxide anion radical detection was established using
the probe. The optimum reaction conditions were pH 8.2, 40 ℃ and 40 min. The conditions of fluorimetric determination
were established as follows: λex = 295 nm, λex = 365 nm and slit width = 5 nm. In the range from 0.4 × 10-6 to 8.0×10-6 mol/L,
relative fluorescence intensity (y) and pyrogallol concentration (x) showed a good linear relationship: y =37.567x + 55.581,
R2 = 0.9834. Superoxide anion radical (O2 - •) scavenging activity of L-ascorbic acid was evaluated using the method and an
IC50 of 0.182 mmol/L was obtained.

Key words: superoxide anion radical, fluorescence probe, synthesis, application