RP-HPLC法测定鼠肝组织儿茶酚氧位甲基转移酶活性的研究  

Study on Catechol-O-Methyltransferase Activity in Rat Liver by RP-HPLC

作　　者：杨丽娜[1] 肖盛元[1] 邓玉林[1] 

机构地区：[1]北京理工大学生命科学与技术学院,北京100081

出　　处：《现代科学仪器》2009年第1期59-61,共3页Modern Scientific Instruments

基　　金：民用航天项目(No:D2220062902)

摘　　要：建立了反相液相色谱法测定鼠肝组织儿茶酚氧位甲基转移酶(COMT)活性的方法。以3,4-二羟基苯甲酸为底物,测定产物4-羟基-3-甲氧基苯甲酸的生成量计算酶活。色谱柱Agilent ZORBAX Eclipse XDB-C18 (150×4.6mm,5μmd),流动相为甲醇/0.7%乙酸水溶液(23/77,v/v),流速1.0mL/min,检测波长254nm,柱温25℃。4-羟基-3-甲氧基苯甲酸的线性范围0.5～20μg/mL,回收率94～98%,RSD小于5%,检测限0.1μg/mL(S/N= 3)。结果表明,该方法简便、快速、重复性好,可应用于鼠肝组织中COMT活性的测定。A reversed phase high performance liquid chromatographic method for determination of Catechol - O - methyltransferase activity in Rat Liver was developed. 4 - Hydroxy - 3 - methoxybenzoic acid was used to de- termine COMT activity when 3,4 - Dihydroxybe - nzoic acid was the substrate. Agilent ZORBAX Eclipse XDB - C18 （ 150 × 4.6mm,5 μmd） was selected. The mobile phase consisted of methol/0.7 % Acetic acid （23/77, v/v） with the flow rate 1.0mL/min at the detection wavelength 254nm. The linear rang of 4 - Hydroxy - 3 - methoxybenzoic acid was from 0.5 to 20μg/mL. The average recovery was 94 -98% with the RSD 〈 5%. The detection limit was 0.1 μg/mL（ S/N = 3 ）. Conclusion： The method was proved to be a simple, quick and good reproducible. The analytic condition could be applied to determination of COMT activity in the rat liver.

关 键 词：RP—HPLC COMT活性 肝组织 4-羟基-3-甲氧基苯甲酸 

分 类 号：R341[医药卫生—基础医学]