过氧亚硝基-鲁米诺化学发光体系的改进  

An Improvement of Chemiluminescent System for Determination of Peroxynitrite Anion

作　　者：范小兵[1] 沙大年[1] 梁小凤[1] 韩超[1] 胡天喜[1] 

机构地区：[1]上海交通大学昂立生物医药研究所,上海200030

出　　处：《生物化学与生物物理进展》2001年第2期251-255,共5页Progress In Biochemistry and Biophysics

摘　　要：建立了一个测定过氧亚硝基阴离子 (ONOO-)化学发光的改进体系 ,测试了某些抗氧化剂清除ONOO-的作用 ,其体系的组成和启动发光的程序如下 :向pH 10 5碳酸缓冲液配的 0 0 1mol/L浓度NaN3 溶液通O330s ,取其 80 0 μl原位注入含有 10 0 μl水配样品和 10 0 μl的 0 0 0 1mol/L鲁米诺溶液中 ,启动化学发光(chemiluminescence ,CL) ,立即测定每 6秒的脉冲数 (CP6S) ,连续测定 10～ 30次 .根据实际需要 ,选其某一次的CL强度作为评判指标 ,对比抗氧化剂的活性 .该发光体系灵敏、简便、且较稳定 ,最低可检测限为 8 74μmol/L的ONOO-量 ,线性范围为 8 74～ 74 0 4μmol/L .批内变异系数 3 35 % (n =10 ) ,批间变异系数 5 5 2 % (n =10 ) .测得维生素C (Vit.C )、茶多酚 (EGCG)、原花菁素、硫脲皆有抑制CL ,即清除ONOO-的作用 .An improvement of chemiluminescent system for determination of peroxynitrite anion (ONOO-) has been made. In this system, the effect of some antioxidants for scavenging ONOO- was tested. The constitute of this system and the program of starting were followed as: The ozone (O-3) was bubbled through a glass-frit into 10 ml 0.01 mol/L solution of sodium azide in CB (pH 10.5) to generate ONOO-. The 800 mul ozonized solution of azide was injected into a glass tude in situ which contains 100 mul sample and 100 mul luminol solutions to initiate chemiluminescence (CL). The pluses /6 seconds (CP6S) were determined immediately and continually for 10 similar to 30 times. A certain CL intensity (CP6S) was chosen as evaluation index to compare the activity of antioxidants. This chemiluminescent system is sensible, simple and stable. The determination limit was 8.74 mu mol/L ONOO-. The linear rang was 8.74 similar to 74.04 mu mol/L ONOO-. The intra batch and inter batch variation coefficient (CV%) of the analysis were 3.35% (n = 10) and 5.52% (n = 10) respectively. It was tested that Vit. C, teapolyphenol, procyanidin and thiourea all have effects on scavenging ONOO-.

关 键 词：化学发光体系 过氧亚硝基阴离子 抗氧化剂筛选 氮自由基 

分 类 号：R965.1[医药卫生—药理学] Q503[医药卫生—药学]