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.
出处
《药学学报》
CAS
CSCD
北大核心
1993年第6期416-421,共6页
Acta Pharmaceutica Sinica
关键词
自由基
高效液相色谱
电化学
检测
HPLC- ECD
Hydroxyl radical
5,5- dimethylpyrroline-1-oxide (DMPO)
EDTAH_2O_2-Fe~(2+0
H_2O2-UV
O_1^-
