A Novel Anthraquinone Derivative as a Chemiluminescence Enhancer in Luminol Assays: Insights Into the CRET Mechanism

Chemiluminescence (CL) reactions are an effective reaction type frequently used in both in vivo and in vitro studies for the determination of many analytes. Some of the molecules used for this purpose are structures that act as enhancers of fluorescence-active molecules, whose activity can be enhanced via chemiluminescence resonance energy transfer (CRET). Such molecules, which are active in terms of fluorescence and operate through the CRET mechanism, are used in combination with molecules such as luminol to produce reaction systems that are more effective. In this study, a model system combining a synthesized anthraquinone derivative with luminol was investigated for its potential use as a CRET-based fluorescent enhancer. Chemical characterization of the molecule was performed using UV, IR, and fluorescence spectroscopy. Additionally, the anthraquinone sample, which was tested in conjunction with luminol as an enhancer on bloodstains, was observed to cause the bloodstains to luminesce for a longer duration and at a higher intensity than when luminol was used alone. As a result, it was observed that unlike conventional luminol enhancers, the synthesized anthraquinone molecule functions as a CRET-based fluorescent acceptor, enabling indirect signal amplification through energy transfer, and could be used as a new type of tracer in biological applications.