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本文采用乙腈除蛋白的方法对血清进行了预处理。维生素D_3与荧光试剂(硫酸-乙醇溶液)的反应条件选择采用均匀设计。荧光分析条件为:激发波长(λ_(ex))427nm,荧光波长(λ_(em))490nm.维生素D_3浓度在0.1～10.0μg/ml范围内与荧光强度成线性关系(相关系数r=0.9981)。人血清中维生素D_3的平均回收率达97.5%以上。精密度也很好,日内变异系数为1.8%(n=6),日间变异系数为4.5%(n=8)以下。并用本法检出服药后血清中维生素D_3的浓度。
In this paper, the method of acetonitrile deproteinization of serum pretreatment. The reaction conditions of vitamin D_3 and fluorescent reagent (sulfuric acid - ethanol solution) were selected using uniform design. Fluorescence analysis conditions were as follows: the excitation wavelength (λ_ (ex)) was 427 nm and the fluorescence wavelength was 490 nm.The linear relationship between the concentration of vitamin D_3 and fluorescence intensity was found in the range of 0.1 ~ 10.0 μg / ml (correlation coefficient r = 0.9981) . The average recovery rate of vitamin D 3 in human serum reached more than 97.5%. The precision was also good, with intraday variability of 1.8% (n = 6) and daytime variability of 4.5% (n = 8). And use this method to detect the serum concentration of vitamin D 3 after taking medicine.