在Tris-HC1缓冲溶液体系中(pH=7.4),研究了1,4-二羟基-2-甲酰基-9,10-蒽醌缩对甲氧基苯基氨基硫脲(EN)与人血清白蛋白(HSA)体系的荧光猝灭光谱和三维荧光光谱,证明EN与HSA可以发生相互作用,使人血清白蛋白的疏水微环境的极性以及构象发生变化.考察了Δλ值、反应介质和离子强度等因素对体系同步荧光光谱特征及强度的影响.在此基础上,建立了以EN为分子探针,运用固定波长同步荧光光谱法测定生物样品中的蛋白质含量的方法.在最佳实验条件下,体系同步荧光强度与HSA在1.380～165.6 mg/L范围内呈良好的线性关系.对11份空白溶液进行平行测定,检出限达到0.414 mg/L,相对标准偏差为1.52％.运用此方法对血清、唾液、尿液进行了加标回收实验,回收率在98.4％～105％.且同步荧光光谱法测定结果与考马斯亮蓝法基本一致.
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