Fluorometric and colorimetric platforms for rapid and sensitive hydroxychloroquine detection in aqueous samples

The detection of pharmaceuticals has been an active area of research with numerous application areas ranging from therapeutic and environmental monitoring to pharmaceutical manufacturing and diagnostics. And, the emergence of COVID-19 pandemic has increased the demand for detection of certain active pharmaceutical ingredients such as Hydroxychloroquine (HCQ) mainly due to their increased manufacturing and usage. In this study, we present two optical, fluorometric and colorimetric, detection platforms for the rapid and sensitive detection of HCQ. These platforms take advantage of the interactions between the highly fluorescent dye Thioflavin T (ThT) and Tel24 G-quadruplex (G4) DNA structure, as well as the salt-induced aggregation behavior of negatively charged citrate-capped silver nanoparticles (Cit-AgNPs) in the presence of HCQ. In the fluorometric method, the addition of HCQ led to a significant and rapid decrease in the fluorescence signal of the ThT + Tel24 probe. In the colorimetric method, HCQ induced the aggregation of Cit-AgNPs in the presence of NaCl, resulting in a noticeable color change from yellowish-gray to colorless. Under the optimized conditions, the colorimetric platform exhibited a linear range of 18.0-240.0 nM and a detection limit of 9.2 nM, while the fluorometric platform showed a linear range of 0.24-5.17 μM and a detection limit of 120 nM. The selectivity of the proposed optical methods towards the target analyte was demonstrated by evaluating the response to other structurally similar small molecules. Finally, the practical applicability of both detection systems was confirmed by analyzing HCQ-spiked human urine samples that yielded average recoveries ranging from 75.4 to 110.2 % for the fluorometric platform and 86.9-98.2 % for the colorimetric platform. These results indicate the potential of the developed methods for HCQ detection in complex matrices.