背景:羟胺(HA)是重要的工业原料和药物中间体。此外,HA是一种重要的细胞代谢产物,它是一氧化氮和氮氧化物形成的中间体。然而,过量的HA对动物和植物都是有毒的。用于检测HA的常规方法是麻烦且复杂的。用荧光探针检测HA方便、灵敏。很少有探针可用于检测羟胺。因此,在这项工作中,开发了一种灵敏和选择性检测HA的荧光探针。
结果:合成了香豆素衍生物SWJT-22作为比色荧光探针,用于检测羟胺(HA),具有较高的灵敏度和选择性。探针对HA的检测限为0.15μM,低于大多数HA探针。在含有SWJT-22的水溶液中加入HA后,溶液的颜色从橙色变为黄色,荧光颜色也从橙色变为绿色。通过1HNMR滴定证实了SWJT-22与HA的反应机理,质谱和圆底烧瓶实验。此外,SWJT-22已制成便携式测试条,用于检测HA。SWJT-22已成功用于细胞成像,可以检测HeLa细胞和RAW264.7细胞中的内源性和外源性HA。
结论:由于羟胺在生物体中的生理作用,选择性和灵敏地检测羟胺至关重要。这项工作为检测羟胺提供了方便的工具,不仅检测细胞中的内源性和外源性HA,而且还制成便携式试纸。HA荧光探针SWJT-22有望促进HA在生理过程中的研究。
BACKGROUND: Hydroxylamine (HA) is vital industrial raw material and pharmaceutical intermediate. In addition, HA is an important cellular metabolite, which is intermediate in the formation of nitric oxide and nitroxide. However, excessive amounts of HA are toxic to both animals and plants. Conventional methods for the detection of HA are cumbersome and complicated. The detection of HA with fluorescent probes is convenient and sensitive. There are few probes available for the detection of
hydroxylamine. Therefore, a fluorescent probe for the sensitive and selective detection of HA was developed in this work.
RESULTS: A coumarin derivative SWJT-22 was synthesized as a colorimetric fluorescent probe to detect hydroxylamine (HA), with high sensitivity and selectivity. The detection limit of the probe to HA was 0.15 μM, which was lower than most probes of HA. Upon the addition of HA to aqueous solution containing SWJT-22, the color of the solution changed from orange to yellow, and the fluorescence color also changed from orange to green. The reaction mechanism of SWJT-22 to HA was confirmed by 1H NMR titrations, mass spectrometry and round bottom flask experiments. Moreover, SWJT-22 had been fabricated into portable test strips for the detection of HA. SWJT-22 had been successfully used in cellular imaging and could detect both endogenous and exogenous HA in HeLa cells and RAW 264.7 cells.
CONCLUSIONS: Due to the physiological role of
hydroxylamine in organisms, it is crucial to detect
hydroxylamine selectively and sensitively. This work provided a convenient tool for the detection of
hydroxylamine, not only to detect endogenous and exogenous HA in cells, but also made into portable test strips. The HA fluorescent probe SWJT-22 is expected to promote the study of HA in physiological processes.