用化学发光法测定辣根过氧化物酶几种体系的研究

Study on Several Systems Used for Chemiluminescent Determination of Horseradish Peroxidase

作者采用四种体系,确定了用化学发光法测定辣根过氧化物酶(HRP)的最佳测定条件和操作步骤。用鲁米诺-H_2O_2(NaOH)可以方便地测定0.05～4pmol HRP,绝对检测限16fmol。用邻苯二酚-H_2O_2(pH6.5)体系可测定0.1～100pmol HRP,绝对检测限3～17fmol。用鲁米诺-H_2O_2-对碘苯酚(pH8.5)体系,可由1分钟时的光强度测定10～50fmol HRP,绝对检测限10fmol。用对羟基联苯代替对碘苯酚,测1或3分钟时的光强度可测定6～40fmol HRP,绝对检测限6.5或3.3fmol.

Four systems were studied used forchemiluminescent datermination of HRPand the determination conditions optimiz-ed. In luminol-H_2O_2(NaOH) system,HRP was determined conveniently in therange of 0.05--4 pmol,with a correlationcoefficient of 0.998 for the linear log-logequation and a limit of detection of 16fmol HRP. In pyrogallol-H_2O_2 (pH 6.5)system, 0.1--100 pmol of HRP were deter-mined, based on three log-log regressionequations with correlation coefficients of0.997, 0.994 and 0.999, respectively, andlimits of datection of 3--17 fmol HRP.Luminol--H_2O_2-para iodophenol system wasused to determine HRP in the range to 10-50 fmol, based on the chemiluminescenceintensity at 1 min and the correlation co-efficient ot the log-log equation being0.996 with a limit of detection of 10 fmolHRP. When p-hydroxybiphenyl was usedto take the place of p--iodophenol and thechemiluminescence intensity at 1 min or3 min measured, 6--40 fmol HRP were de-termined with a correlation coefficient ofo.994 or 0.995 for the log-log equationand a limit of detection of 6.5 or 3.3 fmolHRP.