氧化酶光度法葡萄糖定量测量条件探讨

Study on the experimental conditions for determination of glucose using glucose oxidase and spectrophotometric

依据人体尿糖、血糖的浓度范围，研究从2．0～38．5mmol／L不同浓度的葡萄糖水溶液在不同温度下与葡萄糖氧化酶试剂反应的动力学过程，在可见光谱区，结合分光光度法和最小二乘法对溶液中的葡萄糖含量进行定量分析，分析波长，光谱带宽，反应温度及反应时间等对定量精度的影响。实验结果表明，对于本文所用的葡萄糖氧化酶试剂，在32℃下葡萄糖氧化酶与葡萄糖反应10min后在550nm处进行定量，相关系数超过0．9999，预测标准误差（SEP）0．626mmol／L。如果测量时间和温度偏离指定条件，将产生较大的测量误差；但如果定量波长偏离吸收峰，并在一定带宽（如30nm）范围内进行定量，仍能得到满意的结果。

According to the glucose concentration range in human urine and blood, the dynamic processes of the glucose aqueous solutions between 2.0mmol/L and 38.5mmol/L reacting with glucose oxides （GOD） reagent were studied. The absorption spectra （450 -700nm） in different bandwidth, temperature and time were measured and the glucose concentrations were determined by least square method. In this article, some factors which include the wavelength, bandwidth of the spectrum, the temperature and the reaction time will impact on the determination precision were analyzed. The experimental results show that the optimal measurement conditions of the reagent we used are 32℃, 10 to 15 minutes after reacting and calibrating at 550nm, in this situation, we get the correlation coefficient r 〉 0. 9999, the standard error of the prediction （SEP） is about 0. 626mmol/L. The results also show that if the measurement time and temperature is derivate to this optimal conditions, it will result in a considerable measurement error, however, if we calibrate the glucose concentration within a bandwidth of 20nm rather than lnm at the center wavelength of the absorption spectra peak, the measurement error is small.