Colorimetric pH-sensing of artificial gastric fluid using naphthalimide-based CH acids

In this study, we introduce novel colorimetric pH-sensing probes based on naphthalimide malonate derivatives. These probes were synthesized by reacting 4-bromo-1,8-naphthalimide with various malonates, including malononitrile, ethyl cyanoacetate, and diethyl malonate. Each derivative exhibited distinct pH-sensing characteristics due to their differing CH acidities. The malononitrile-based probe, NPI-N2, demonstrated pronounced chromogenic pH-signaling behavior, transitioning from colorless to red-violet, accompanied by a decrease in fluorescence intensity. Notably, NPI-N2 retained its pH-sensing capability in the presence of common metal ions, anions, and pepsin, a key component of gastric fluid. The pK_a of NPI-N2 was determined to be 3.08 through pH-dependent absorbance curve fitting. To modulate the pH-sensing range, ester-nitrile (NPI-EN) and diethyl ester (NPI-E2) subunits were incorporated into the naphthalimide framework, resulting in increased pK_a values of 6.73 and 10.76, respectively. The pH-signaling mechanism of NPI-N2 was elucidated by ¹H and ¹³C NMR spectroscopy, revealing deprotonation of the malononitrile moiety and subsequent resonance extension through the naphthalimide structure. To facilitate practical pH determination, NPI-N2 was integrated into a paper-based test strip, enabling convenient and reliable pH measurement of artificial gastric fluid.