1,3,5,7-四甲基-2,6-二乙酯基-8-(3’,4’-二氨苯基)-二氟化硼-二吡咯甲烷荧光光度法灵敏测定血液中亚硝基硫醇

Sensitive Determination of S-Nitrosothiols in Blood by Spectrofluorimetry using 1,3,5,7-Tetramethyl-2,6-dicarbethoxy-8-(3',4'-diaminophenyl)-Difluoroboradiaza-s-indacence as Probe

S-亚硝基硫醇在生物体内具有传递信号、改变蛋白质的结构和功能、阻止有害物质对细胞的伤害等重要的生理作用。建立了一种灵敏、简单的应用荧光探针1,3,5,7-四甲基-2,6-二乙酯基-8-(3’,4’-二氨苯基)-二氟化硼-二吡咯甲烷标记并测定S-亚硝基硫醇的荧光光度法:在1.0×10-4mol/LHg2+存在下,荧光探针与S-亚硝基硫醇在30℃反应15min生成衍生物,在激发和发射波长分别为500nm和510nm处测定其荧光强度。荧光强度和S-亚硝基硫醇的浓度在(8.0～800.0)×10-9mol/L范围内成良好的线性关系。当信噪比等于3时,方法的检出限为6.0×10-10mol/L。该方法对健康人与心血管疾病患者血液中S-亚硝基硫醇的测定的加标回收率为97.46%～103.10%。

S-nitrosothiols （RSNO） is an extremely important and versatile messenger in the biological system. It can transport signal,change the structure and function of protein and prevent damage of deleterious substance to cells in vivo. In this work,a spectrofluorimetric method for determining RSNO is proposed. An excellent fluorescence probe 1,3,5,7-tetramethyl-2,6-dicarbethoxy-8-（3 ,4 -diaminophenyl）-difluoroboradiaza-s-indacence is used to label RSNO. The derivatization reaction performs in aqueous medium at 30℃ for 15min in the presence of 1.0 × 10-4 mol/L Hg^2＋ . The fluorescence of the derivative is detected at an emission wavelength of 510nm with excitation at 500 nm. The response is a linear for concentration in the range of （8.0 - 800.0） × 10^-9 mol/L with a correlation coefficient of 0.9990. The detection limit is 6.0 × 10^-10 mol/L（ S/N = 3）. The method is applied to determine RSNO in the blood of healthy persons and patients suffering from cardiovascular diseases. Recoveries of RSNO from spiked blood samples are between 97.46% and. 103.10%. The results indicate that the as-presented method is rapid, simple, sensitive and feasible.