用高效液相电化学检测直接测定氧自由基  

DIRECT MEASUREMENT OF OXY-RADICALS USING HPLC-ECD

作　　者：刘雨[1] 张均田[1] 

机构地区：[1]中国医学科学院药物研究所,北京100050

出　　处：《药学学报》1993年第6期416-421,共6页Acta Pharmaceutica Sinica

摘　　要：应用DMPO捕捉羟自由基(·OH)生成DMPO—羟游离基(DMPO-OH),经高效液相电化学检测羟自由基。选用ODS反相柱(10 μm)及柠檬酸30 mmol/L—乙酸钠50 mmol/L—3%乙腈为流动相,流速1-2 ml/min,检测电压0.6 V。采用EDTA-H_2O_2-Fe^(2+)(FeSO_4300 nmol/L,EDTA 300μmol/L,H_2O_2180μmol/L及DMPO 2 mmol/L和H_2O_2光照(H_2O_218 mmol/L,DMPO 2 mmol/L光照6 min)两种产生·OH的体系作为药物筛选及作用机制探讨。其RSD分别为6.1和8.0%。检测灵敏度和ESR相似,本文介绍了O_2的检测方法。A direct, sensitive, simple and specific high-pressure liquid chromatographic (HPLC) method was used for the quantitation of hydroxyl radicals by means of oxy-radical trapping of DMPO to form DMPO-OH adducts. The DMPO-OH adduct peak was separated successfully and identified by HPLC-ECD with a Waters ODS reversedphase 10 μm column. The mobile phase composed of citric acid/sodium acetate (citric acid—30 mmol/L—sodium acetate 50 mmol/L—3% acetonitrile, pH 5. 1), at a flow rate of 1.2 ml/min and detection potential of 0.6 V with Ag/AgCl as reference electrode. Both EDTA—Fe^(2+)-H_2O_2 (FeSO_4300 μmol/L, EDTA 300 μmol/L, H_2O_2180 μmol/L and DMPO 2 mmol/L) and H_2O_2 photolysis (H_2O_218 mmol/L and DMPO 2 mmol/L photolysis for 6 min) systems were taken to produce hydroxyl free radicals for screening new drugs and studyins the mechanism of action. The relative standard deviations were 6.1 and 8.0% respectively. The sensitivity of the method was shown to be similar to that of ESR. The method for detection of superoxide anions with HPLC-ECD was also described.

关 键 词：自由基 高效液相色谱 电化学 检测 

分 类 号：R914.1[医药卫生—药物化学]