A highly sensitive and colorimetric fluorescent probe for visualizing hydroxylamine in immune cells

Area of Science:

• Chemical Biology
• Analytical Chemistry
• Biochemistry

Background:

• Hydroxylamine (HA) is a crucial industrial and pharmaceutical intermediate, and a cellular metabolite.
• Excessive HA levels are toxic to organisms, necessitating accurate detection methods.
• Existing HA detection methods are often cumbersome, highlighting the need for sensitive and selective probes.

Purpose of the Study:

• To develop a novel fluorescent probe for sensitive and selective detection of hydroxylamine (HA).
• To characterize the probe's performance and reaction mechanism.
• To demonstrate the probe's utility in cellular imaging and portable detection.

Main Methods:

• Synthesis of coumarin derivative SWJT-22 as a colorimetric fluorescent probe.
• Spectroscopic analysis (UV-Vis, fluorescence) to determine probe response to HA.
• ¹H NMR titrations and mass spectrometry to elucidate the reaction mechanism.
• Fabrication of portable test strips and application in cellular imaging (HeLa and RAW 264.7 cells).

Main Results:

• SWJT-22 exhibited high sensitivity and selectivity for HA detection, with a low detection limit of 0.15 μM.
• Colorimetric change from orange to yellow and fluorescence shift from orange to green upon HA addition.
• Successful application in detecting endogenous and exogenous HA in various cell lines.
• Development of portable test strips for practical HA detection.

Conclusions:

• The developed fluorescent probe SWJT-22 offers a convenient and effective tool for selective and sensitive HA detection.
• SWJT-22 facilitates the study of HA's physiological roles by enabling intracellular detection and portable analysis.
• This probe is expected to advance research on hydroxylamine in biological processes.