引用本文:
耿韶光, 江晓莹, 崔凤灵, 时蕾. 一种缩氨基硫脲类衍生物探针测定生物样品中的蛋白质含量[J]. 应用化学,
2013, 30(5): 555-559.
doi:
10.3724/SP.J.1095.2013.20312
Citation: GENG Shaoguang, JIANG Xiaoying, CUI Fengling, SHI Lei. Determination of Protein in Biological Samples Using a Thiosemicarbazone Derivative as a Fluorescence Probe[J]. Chinese Journal of Applied Chemistry, 2013, 30(5): 555-559. doi: 10.3724/SP.J.1095.2013.20312
Citation: GENG Shaoguang, JIANG Xiaoying, CUI Fengling, SHI Lei. Determination of Protein in Biological Samples Using a Thiosemicarbazone Derivative as a Fluorescence Probe[J]. Chinese Journal of Applied Chemistry, 2013, 30(5): 555-559. doi: 10.3724/SP.J.1095.2013.20312
一种缩氨基硫脲类衍生物探针测定生物样品中的蛋白质含量
摘要:
在Tris-HCl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△λ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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关键词:
- 蒽醌缩对甲氧基苯基氨基硫脲
- / 人血清白蛋白
- / 三维荧光光谱
- / 同步荧光光谱
English
Determination of Protein in Biological Samples Using a Thiosemicarbazone Derivative as a Fluorescence Probe
Abstract:
The interaction between(E)-2((1,4-dihydroxy-9,10-anthraquinone-2-yl) methylene)-N-(4-methoxyphenyl) hydrazinecarbothioamide (EN) and human serum albumin (HSA) is investigated by fluorescence in Tris-HCl buffer solution (pH=7.4). The results of fluorescence experiments and three-dimensional fluorescence spectra indicate that the presence of EN can change the polarity of the hydrophobic microenvironment and induce minor changes of the secondary structure of HSA. Under the optimum conditions,the linear range of the protein is 1.380~165.6 mg/L and the detection limit is 0.414 mg/L with the synchronous fluorescence method.The contents of the proteins in human serum,saliva and urine samples are detected and the recovery is ranged from 98.4% to 105%. The effects of coexistent material are also investigated. The results of synchronous fluorescence method are in good agreement with the Coomassie Brilliant Blue(CBB) method.
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