Theone-electron reduced and protonatedcongener of nitric oxide (NO), nitroxyl (HNO) have been demonstrated withexcellent bio-pharmacological effects in cardiovascular disorder treatment,which are distinctive from that of NO. The high reactivity of HNO challenges theaccurate detection. To resolve this problem, a near-infrared (NIR) metal-freefluorescent probe, ER-JN, has been developed for the detection of intracellularHNO concentration in simulated physiological conditions and living cells. Theprobe is consisted of two moieties, the aza-BODIPY fluorophore and the HNOrecognition unit, diphenylphosphinobenzoyl group. The probe is purified by columnchromatography on silica eluting with CH₂Cl₂ to give theproduct as a green solid. Yield: 28 %. Ourprobe exhibits high sensitivity, good selectivity, and low cytotoxic effect,and can be applied to the fluorescent bio-imaging of HNO in simulated physiologicalconditions. When detected HNO, quantum yield increases from 0.01 to 0.35. The linearrange locate at 0~50 µmol/L. The detection limit is determined to be 0.03 μmol/L(S/N= 3). Using confocal laser scanning microscope imaging analysis, the probe can detectHNO concentration changes in living cells. Furthermore, the results of flowcytometry confirm that our probe can be employed for the qualitative andquantitative detection of intracellular HNO concentration level. In this work,we have found that probe ER-JN not only detects HNO in aqueous and in livingcells, but also targets endoplasmic reticulum. We anticipate that ER-JN willprovide experimental bases in studying physiological and pathological functionsof HNO in cells, in vitro and in vivo.

A Near-infraredfluorescent probe for the detection of HNO in living cells

Ping Liu¹, Xiaoyue Han^1,2, Fabiao Yu¹*,Lingxin Chen¹*

CHINESE JOURNAL OF ANALYTICAL CHEMISTRY Volume 43, Issue 12, December 2015

Chin J Anal Chem, 2015, 43(12), 1829–1836.

刘萍, 韩潇玥, 于法标, 陈令新. 用于检测细胞中亚硝酰氢的近红外荧光探针及其生物应用[J]. 分析化学, 2015, 43(12): 1829-1836