Department of Medicinal and Applied Chemisrty, Kaohsiung Medical University, Kaohsiung 807, Taiwan.
Anal Chim Acta. 2011 Dec 5;708(1-2):141-8. doi: 10.1016/j.aca.2011.10.005. Epub 2011 Oct 8.
A colorless and non-fluorescent rhodamine derivative, rhodamine B hydrazide (RH), is applied to detect nitric oxide and form fluorescent rhodamine B (RB). The reaction mechanism of RH with NO is proposed in this study. The probe shows good stability over a broad pH range (pH>4). Furthermore, fluorescence intensity of RH displays an excellent linearity to the NO concentration and the detection limit is as low as 20 nM. A 1000-fold fluorescence turn-on from a dark background was observed. Moreover, the selectivity study indicated that the fluorescence intensity increasing in the presence of NO was significantly higher than those of other reactive oxygen/nitrogen species. In exogenously generated NO detection study, clear intracellular red fluorescence was observed in the presence of S-nitroso-N-acetyl-D,L-penicillamine (SNAP, a kind of NO releasing agent). In endogenously generated NO detection study, increasing incubation time of RH with lipopolysaccharied (LPS) pre-treated cells could obtain a highly fluorescent cell image. These cell imaging results demonstrated that RH can efficiently penetrate into Raw 264.7 cells and be used for detection of exogenously and endogenously generated nitric oxide.
一种无色非荧光的罗丹明衍生物，罗丹明 B 酰肼（RH），被应用于检测一氧化氮并形成荧光罗丹明 B（RB）。本研究提出了 RH 与 NO 的反应机制。该探针在较宽的 pH 范围内（pH>4）具有良好的稳定性。此外，RH 的荧光强度对 NO 浓度表现出极好的线性关系，检测限低至 20 nM。从暗背景观察到 1000 倍的荧光开启。此外，选择性研究表明，在存在 NO 的情况下，荧光强度的增加明显高于其他活性氧/氮物质。在体外生成的 NO 检测研究中，在 S-亚硝基-N-乙酰-D,L-青霉胺（SNAP，一种 NO 释放剂）存在的情况下，观察到细胞内明显的红色荧光。在体内生成的 NO 检测研究中，增加 RH 与脂多糖（LPS）预处理细胞孵育时间可以获得高荧光细胞图像。这些细胞成像结果表明，RH 可以有效地穿透 Raw 264.7 细胞，并用于检测外源性和内源性产生的一氧化氮。
