A deoxygenation-switch-based red-emitting fluorogenic light-up probe for the detection of highly toxic free bilirubin in human blood serum

Abstract

Reaction-based chemical switches are attracting great interest due to their high selectivity, and their use has become a powerful technique for developing fluorogenic probes. Herein, a benzorhodol-derivative-attached N-oxide probe (DEBNox) has been designed as a new fluorogenic probe for the detection of the biologically toxic species bilirubin based on a deoxygenation switching mechanism. Upon reaction with added Fe³⁺, bilirubin produces Fe²⁺ ions in situ, which in turn promote a deoxygenation reaction with DEBNox to generate the corresponding high-red-fluorescence (λ_em: ∼623 nm) benzorhodol derivative (DEB). This type of Fe³⁺-mediated response helps the probe to act as a qualified turn on selective fluorescence sensor for bilirubin with a detection range as low as 33 nM. Moreover, the probe was successfully employed to detect free bilirubin in human blood serum specimens with acceptable accuracy and reliability. This DEBNox-based light-up strategy also facilitates the construction of reliable and highly sensitive assays based on a paper-based strategy, similar to pH-indicator paper, as is demonstrated here via bilirubin detection in real serum samples. These findings could be useful for developing powerful diagnostic tools for the detection of free bilirubin in the near further.