A phenothiazine-based ratiometric fluorescence probe for the detection of hydroxylamine in real water and living cells

Abstract

In this study, a phenothiazine-based ratiometric fluorescent probe PCHO was developed for highly sensitive and specific detection of hydroxylamine (HA). In the presence of HA, the aldehyde group on the PCHO molecule underwent a specific nucleophilic addition with HA to form an oxime group, accompanied by significant changes in fluorescence from green to blue. This detection mechanism was well supported by ¹H NMR titration, HRMS and DFT calculations. The probe PCHO exhibited high sensitivity for HA detection (LOD was 0.19 μM) with a rapid response time (1 min), high selectivity and strong anti-interference performance. Surprisingly, the probe PCHO could selectively distinguish HA from its similar competing agents such as hydrazine and amines. Moreover, paper strips loaded with PCHO were prepared and combined with a smartphone to achieve point-of-care and visual detection of HA. The probe PCHO was further applied for the detection of HA in real water samples, achieving a recovery rate of 98.90% to 104.86% and an RSD of 0.86% to 2.44%, confirming the accuracy and reliability of the method. Additionally, the probe PCHO was used for imaging analysis of HA in living cells, providing a powerful visualization tool for exploring the physiological functions of HA in vivo.