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一氧化氮(NO)是生物体内的一种重要生物信号传导分子,广泛参与生物体内多种生理及病理过程。为建立快速高效、准确检测生物体释放NO的分析方法,本文选用高灵敏度、高选择性的NO特异性荧光探针8-(3,4-diaminophenyl)-2,6-bis(2-car-boxyethyl)-4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene(DAM-BO-PH),激发波长和发射波长分别为520 nm和535 nm,以高通量微孔板(384孔)作为实验工具载体。该方法荧光强度与NO浓度在8.0×10-10~8.0×10-7mol.L-1范围内呈良好线性关系,R=0.9989,检出限为0.18 nmol.L-1,回收率为98%~102%。该方法应用于多种生物样品中释放NO的分析检测,结果令人满意。
Nitric oxide (NO) is an important biological signal transduction molecule in organism, which is widely involved in many physiological and pathological processes in the organism. In order to establish a rapid, efficient and accurate method for the detection of NO released by organism, a high sensitivity and high selectivity NO-specific fluorescent probe 8- (3,4-diaminophenyl) -2,6-bis (2- boxyethyl 4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a and 4a-diaza-s-indacene with the excitation and emission wavelengths of 520 nm and 535 nm, with high-throughput microplate (384 wells) as the experimental tool carrier. The fluorescence intensity of this method showed a good linear relationship with the concentration of NO in the range of 8.0 × 10-10 ~ 8.0 × 10-7mol·L-1, R = 0.9989, the detection limit was 0.18 nmol.L-1, the recovery was 98% ~ 102%. The method has been applied to the analysis and detection of NO release in a variety of biological samples with satisfactory results.