Detection of Hydrogen Peroxide in vitro and in vivo Using Peroxalate Chemiluminescent Micelles

Abstract

Hydrogen peroxide plays a key role as a second messenger in the normal cellular signaling but its overproduction has been implicated in various life-threatening diseases. Peroxalate chemiluminescence is the light emission from a three component reaction between peroxalate, hydrogen peroxide and fluorophores. It has proven great potential as a methodology to detect hydrogen peroxide in physiological environments because of its excellent sensitivity and specificity to hydrogen peroxide. We developed chemiluminescent micelles composed of amphiphilic polymers, peroxalate and fluorescent dyes to detect hydrogen peroxide at physiological concentrations. In this work, we studied the relationship between the chemiluminescence reactivity and stability of peroxalate by varying the substitutes on the aryl rings of peroxalate. Alkyl substitutes on the aryl ring of peroxalate increased the stability against water hydrolysis, but diminished the reactivity to hydrogen peroxide. Chemiluminescent micelles encapsulating diphenyl peroxalate showed significantly higher chemiluminescence intensity than the counterpart encapsulating dimethylphenyl or dipropylphenyl peroxalate. Diphenyl peroxalate-encapsulated micelles could detect hydrogen peroxide generated from macrophage cells stimulated by lipopolysaccharide (LPS) and image hydrogen peroxide generated during LPS-induced inflammatory responses in a mouse.

Possibilities and Challenges for Quantitative Optical Sensing of Hydrogen Peroxide

“…There are also reports on chemiluminescence-based H 2 O 2 sensitive NPs, which consist of a fluorescent dye, incorporated in a peroxalate ester containing polymer [98] or a dye and the peroxalate ester incorporated in a polymeric micelle [99]. The peroxalate ester reacts with H 2 O 2 forming a highly energetic dioxetanedione, which-upon collision-transfers its energy to the dye molecule and degrades into two CO 2 molecules.…”

Oxalate-curcumin–based probe for micro- and macroimaging of reactive oxygen species in Alzheimer’s disease

“…When oxalate is mixed with H 2 O 2 , it converts into 1,2-dioxetanedione, which is not stable and instantly decomposes to generate CO 2 and release photons (35,36). Nanoparticles containing oxalate and dyes have been used to detect H 2 O 2 in vitro and in vivo via chemiluminescence (37)(38)(39). Our design strategy is different from chemiluminescent nanoparticles, which use oxalate as an exciting source for a fluorophore that is confined inside the nanoparticle.…”

Hydrogen peroxide-responsive copolyoxalate nanoparticles for detection and therapy of ischemia–reperfusion injury

“…As shown in Fig. 3b, the addition of H 2 O 2 to the suspension of rubrene-loaded HPOX nanoparticles (HPOX/ Rb) instantaneously initiated chemiluminescence reactions to produce high energy intermediate dioxetanedione that chemically excites Rb, leading to the generation of strong light emission at 565 nm [17,18]. The nanoparticles showed no light emission without H 2 O 2 .…”