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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >Boron-chelating fluorescent probe (BOPB) in the red region combined with CE-LIF for the detection of NO in mice liver
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Boron-chelating fluorescent probe (BOPB) in the red region combined with CE-LIF for the detection of NO in mice liver

机译:红色区域的硼螯合荧光探针(BOPB)结合CE-LIF检测小鼠肝脏中的NO

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摘要

Precise measurement of nitric oxide (NO) is of great importance to understand the function of NO in liver and the mechanism of liver injury. 8-(3', 4'-Diamino phenyl)3,5-(2-hydroxyphenyl)-dimethylene pyrrole (BOPB), a fluorescent probe in the red region (>600 nm) newly developed in our group, has good photostability and excitation/emission wavelength of 622/643 nm matching well with commercial 635 nm semiconductor laser of CE-LIF detection. Therefore, BOPB was used in CE-LIF for the determination of NO in mice liver. Both derivatization and separation conditions were optimized. Derivatization reaction of BOPB and NO was carried out in pH 7.4 PBS buffer at 35 degrees C for 12 min and the separation of NO derivative of BOPB (BOPB-T) was achieved within 7.0 min in pH 9.0 running buffer containing 15 mM H3BO3-NaOH and 15 mM SDS. Good linearity was found in the range of 1.0 x 10(-9)-5.0 x 10(-7) M with the LOD of 0.02 nM. The proposed method was applied to the analysis of NO in real samples, and NO concentration was obviously increased in acute liver injury of mice. Compared to existing derivatization-based CE-LIF methods for NO, this method has lower LOD and less background interference owing to detection wavelength of BOPB in the red region.
机译:一氧化氮(NO)的精确测量对于了解NO在肝脏中的功能以及肝损伤的机制非常重要。 8-(3',4'-二氨基苯基)3,5-(2-羟基苯基)-二亚甲基吡咯(BOPB)是我们小组新开发的红色区域(> 600 nm)的荧光探针,具有良好的光稳定性和622/643 nm的激发/发射波长与CE-LIF检测的商用635 nm半导体激光器非常匹配。因此,BOPB被用于CE-LIF测定小鼠肝脏中的NO。优化了衍生化和分离条件。 BOPB和NO的衍生反应在pH 7.4 PBS缓冲液中于35摄氏度进行12分钟,并在7.0 min内在pH 9.0含15 mM H3BO3-NaOH的运行缓冲液中分离BOPB的NO衍生物(BOPB-T)和15 mM SDS。发现良好的线性范围为1.0 x 10(-9)-5.0 x 10(-7)M,LOD为0.02 nM。该方法用于实际样品中NO的分析,小鼠急性肝损伤中NO浓度明显升高。与现有基于NO的基于衍生化的CE-LIF方法相比,该方法具有较低的LOD,并且由于红色区域中BOPB的检测波长而具有较少的背景干扰。

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