葡萄糖氧化酶-过氧化物酶改良法的建立——血糖测定时间窗口的探讨

Establishment of improved glucose oxidase-peroxide enzyme method:time window for measurement of blood glucose

目的对小鼠血糖葡萄糖氧化酶-过氧化物酶(glucose oxidase-peroxide enzyme,GOD-POD)法进行改良。方法针对糖尿病模型大鼠的血糖测定,优化了影响测定结果的一些重要因素(如GOD-POD浓度、显色反应稳定时段、波长、线性范围等)。结果 (1)GOD-POD浓度的选择:当两者的浓度均为1040IU·ml-1,有一显色反应相对稳定的时段(室温下反应从90min至180min),此时样品光密度值(optical density,OD)的变异系数(coefficient of variation,CV)较小(0.846%±0.205%,n=3),可用作对较大样本血糖测定的时间窗口;(2)波长的选择:GOD-POD浓度均为1040 IU·ml-1时,550nm的测定灵敏度比505nm好;(3)线性范围:在葡萄糖浓度为3.75～480mg·dl-1范围内线性关系良好,最低检测值达3.75mg·dl-1。结论相对于原法,改良法适于糖尿病模型大鼠的血糖测定及降糖药物的效应观察。

Aim To improve a glucose oxidase-peroxide enzyme（GOD-POD） method.Methods Some important factors that have influence on accurate measurement of blood glucose have been optimized.Results Firstly,the choice of GOD and POD concentrations：when the concentrations of both GOD and POD were 1 040 IU·ml-1 in color reactions,there was a relatively stable time window（90 ～ 180 min under room temperature） for the detection of optical density（OD） during which the coefficient of variation（CV） of OD values was the smallest（0.846% ± 0.205%,n = 3）.Secondly,the choice of wavelength：550 nm wavelength was better than 505 nm in sensitivity when the GOD-POD concentration was 1 040 IU · ml-1.Thirdly,the range and limit of detection：the range of linearity was from 3.75 mg·dl-1 to 480 mg·dl-1 with a value for r = 0.998;the limit of detection was 3.75 mg·dl-1.Conclusion Compared with the primary method,the improved GOD-POD method can be suitable to measure the blood glucose of diabetic rats and evaluate the effects of drugs for decreasing blood glucose.